摘要:ObjectiveTo observe the effects of modified Chaihu Shugansan on the calmodulin-dependent protein kinase Ⅱ(CaMKⅡ)/cAMP-response element binding protein (CREB) signaling pathway in the hippocampus and heart tissue of a rat model with myocardial ischemia and depression and explore the mechanism by which this formula prevents and treats coronary heart disease combined with depression.MethodsThe model of myocardial ischemia combined with depression was established by high-fat diet, intraperitoneal injection of isoproterenol (ISO), and chronic unpredictable mild stress (CUMS). A total of 108 SD male rats were randomly divided into normal group, model group, high (23.4 g·kg-1), medium (11.7 g·kg-1), and low (5.85 g·kg-1) dose groups of modified Chaihu Shugansan, CaMKⅡ inhibitor (KN93) group, and KN93 + high, medium, and low dose groups of modified Chaihu Shugansan, with 12 rats in each group. From the first day of modeling to the end of modeling, drugs were administered once a day. In the seventh and eighth weeks, the KN93 group and the KN93 + high, medium, and low dose groups of modified Chaihu Shugansan were intraperitoneally injected with KN93 three times weekly. At the end of the eighth week, behavioral tests including sucrose preference, open field, and elevated plus maze were conducted. Electrocardiogram (ECG) lead Ⅱ changes were observed in each group of rats, and hematoxylin-eosin (HE) staining was performed to observe changes in heart tissue. Serum levels of triglycerides (TG), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), and lactate dehydrogenase (LDH) were measured by using an enzyme-labeled instrument. Creatine kinase (CK) and creatine kinase-MB (CK-MB) were detected by ultraviolet spectrophotometry, while serum monocyte chemoattractant protein-1 (MCP-1) was measured by enzyme-linked immunosorbent assay (ELISA). Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect mRNA expression of CaMKⅡ and CREB in hippocampal and heart tissue, and Western blot was performed to assess protein expression of CaMKⅡ, phosphorylated (p)-CaMKⅡ, CREB, and p-CREB.ResultsCompared to the normal group, the model group showed significant reductions in sucrose preference rate, total activity distance in the open field, number of entries into the center area of the open field, and percentage of entries into the open arms of the elevated plus maze (P<0.01). The ECG showed ST-segment elevation, and HE staining showed serious degeneration of myocardial fibers, disordered arrangement, and infiltration of a large number of inflammatory cells. In addition, serum TC and LDL levels increased (P<0.01), and HDL level decreased (P<0.01). CK, CK-MB, LDH, and MCP-1 levels significantly increased (P<0.05, P<0.01). The mRNA expression of CaMKⅡ and CREB and the protein expression of p-CaMKⅡ and p-CREB decreased in the hippocampal tissue (P<0.05, P<0.01), but those increased in the heart tissue (P<0.01). Compared to the model group, the high, medium, and low dose groups of modified Chaihu Shugansan showed improvements in these abnormalities. The KN93 group had reduced sucrose preference, total activity distance in the open field, number of entries into the center area of the open field, and percentage of entries into the open arms of the elevated plus maze (P<0.01), as well as decreased serum CK, CK-MB, LDH, and MCP-1 levels (P<0.05, P<0.01). KN93 also reduced ST-segment elevation, alleviated the degeneration degree of myocardial fibrosis, and lowered inflammatory cell infiltration. The mRNA expression of CaMKⅡ and CREB and the protein expression of p-CaMKⅡ and p-CREB in both the hippocampal and heart tissue were reduced (P<0.05, P<0.01). The KN93 + high, medium, and low dose groups of modified Chaihu Shugansan showed further improvements in these abnormalities compared to the KN93 group.ConclusionThe modified Chaihu Shugansan exerts antidepressant and myocardial protective effects in rats with myocardial ischemia and depression, possibly related to bidirectional regulation of the CaMKⅡ/CREB signaling pathway, with the high-dose modified Chaihu Shugansan showing the best effects.
关键词:modified Chaihu Shugansan;myocardial ischemia combined with depression;calmodulin-dependent protein kinaseⅡ (CaMKⅡ)/cAMP-response element binding protein (CREB) signaling pathway
摘要:ObjectiveTo explore the application value of modified Wendantang in the treatment of perimenopausal insomnia of phlegm heat type and observe its effects on neurotransmitters and body fat.MethodsA total of 162 patients with perimenopausal insomnia of phlegm heat type who were treated in the Traditional Chinese Medicine Department of The First Affiliated Hospital with Nanjing Medical University from May 2021 to November 2023 were randomly divided into two groups before inclusion in the study. The control group of 81 cases received conventional Western medicine treatment, while the observation group of 81 cases received combined treatment with modified Wendantang, and the treatment lasted for 4 weeks.ResultsInterleukin-β (IL-1β), and tumor necrosis factor alpha (TNF-α) decreased, while serum dopamine (DA), melatonin (MT) and serotonin (5-HT) increased in both groups after treatment compared with before treatment (P<0.05). After treatment, compared with the control group, the observation group showed better improvement in the above indicators (P<0.05). The levels of follicle stimulating hormone (FSH) and luteinizing hormone (LH) decreased, while the level of estradiol (E2) increased in both groups after treatment compared with before treatment (P<0.05). Compared with the control group, the observation group showed better improvement in the above sex hormone indicators after treatment (P<0.05). The rapid eye movement sleep period (REM) and total sleep time (TST) were prolonged, and the sleep latency (SL) was shortened in the two groups after treatment compared with before treatment (P<0.05). Compared with the control group, the observation group showed better improvement in sleep conditions after treatment (P<0.05). The Pittsburgh sleep quality index (PSQI) and Traditional Chinese Medicine (TCM) total symptom score decreased significantly in both groups after treatment compared with before treatment (P<0.05). Compared with the control group, the observation group showed better improvement in PSQI and TCM total symptom score after treatment (P<0.05). The total effective rate of the observation group was 93.83% (76/81), while that of the control group was 82.72% (67/81), and the two had a statistically significant difference (χ2=9.135, P<0.05).ConclusionFor patients with perimenopausal insomnia of phlegm heat type, the application of modified Wendantang can help regulate neurotransmitters and inflammatory factor levels, improve sex hormone levels, enhance sleep quality, and improve therapeutic effects.
摘要:ObjectiveTo elucidate the correlation between alterations in digestion and absorption functions and hepatic deficiency states in aging rats based on the R-spondin1/Wnt3a signaling pathway, and reveal the intervention mechanism of Bugansan.MethodsForty-eight SPF-grade male SD rats were randomly assigned to six groups: blank control, model, low-, medium-, and high-dose (7.03, 14.06, 28.12 g·kg-1, respectively) Bugansan, and vitamin E (suspension, 27 mg·kg-1), with 8 rats in each group. The rat model of aging was established by intraperitoneal injection of D-galactose (400 mg·kg-1), while the blank control group was injected with normal saline. Since the day of modeling, rats in intervention groups received corresponding agents by gavage, and those in blank control and model groups received an equal volume of normal saline (10 mL·kg-1). General biological features such as fur color, activity, body mass, water intake, and food intake were observed. Meanwhile, the content of malondialdehyde (MDA) and the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the serum were measured to assess aging. Grip strength and the content of total bile acids (TBA) and the activity of α-amylase (AMY) in the serum were measured to evaluate hepatic deficiency states. The activity of β-galactosidase (β-gal) in the duodenum was measured to evaluate intestinal senescence. The levels of glucagon-like peptide-1 (GLP-1), vasoactive intestinal peptide (VIP), and D-xylose in the serum were determined to assess digestion and absorption functions of the small intestine. Hematoxylin-eosin staining was conducted to observe pathological changes of the duodenum to assess the small intestine damage. Immunohistochemical staining was employed to visualize the expression of B-cell-specific Moloney murine leukemia virus integration site 1 (Bmi1) and leucine-rich repeat-containing G protein-coupled receptor 5 (Lgr5) in the duodenal tissue. Moreover, Real-time quantitative polymerase chain reaction (Real-time PCR) was utilized to quantify the mRNA levels of Ki67, Bmi1, and Lgr5 to assess proliferation and regeneration of the small intestine. Additionally, the mRNA levels of R-spondin1, Wnt3a, β-catenin, and glycogen synthase kinase-3β (GSK-3β) and the protein levels of R-spondin1, Wnt3a, β-catenin, and phosphorylated GSK-3β (p-GSK-3β) in the duodenum were determined by Real-time PCR and Western blot, respectively, to analyze the mechanisms of intestinal digestion and absorption dysfunction in aging rats and the regulatory characteristics of Bugansan.ResultsCompared with blank control group, the model group showed decreases in body mass, water intake, food intake, grip strength, activities of SOD, GSH-Px, and AMY in the serum and content of GLP-1, VIP and D-xylose in the serum (P<0.05), increases in the content of MDA and TBA in the serum and β-gal activity in the duodenum (P<0.05), reductions in villus length, villus width, crypt depth, and villi/crypt (V/C) value, down-regulated mRNA and protein levels of Ki67, Lgr5, Bmi1, R-spondin1, Wnt3a, β-catenin, and up-regulated level of GSK-3β, phosphorylation (p)-GSK-3β (P<0.05). Compared with the model group, Bugansan increased the body mass, water intake, food intake, grip strength, and activities of SOD, GSH-Px, and AMY and levels of GLP-1, VIP and D-xylose in the serum (P<0.05), while decreasing the content of MDA and TBA in the serum and β-gal activity in the duodenum (P<0.05). Furthermore, Bugansan increased the villus length, villus width, crypt depth, and V/C value, up-regulated the mRNA and protein levels of Ki67, Lgr5, Bmi1, R-spondin1, Wnt3a, β-catenin, and down-regulated the level of GSK-3β and p-GSK-3β (P<0.05).ConclusionAging rats exhibit obvious impairments in digestion and absorption functions, accompanied by a state of hepatic deficiency. The traditional Chinese medicine approach of tonifying liver Qi effectively ameliorates aging-related changes by modulating the R-spondin1/Wnt3a signaling pathway to mitigate intestinal senescence and enhance digestion and absorption functions, ultimately contributing to the delay of aging.
关键词:aging;digestion and absorption dysfunction;R-spondin1/Wnt3a signaling pathway;liver Qi deficiency;Bugansan
摘要:ObjectiveBased on the TCM theory of "Yang transforms materials to Qi while Yin constitutes material form", this paper explored the effects of Zuogui Wan and Yougui Wan on the molecular mechanism of mitochondrial biogenesis during the adipogenic differentiation process of rat bone marrow mesenchymal stem cells (BMSCs) by mediating peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) and peroxisome proliferators-activated receptor γ (PPARγ), providing theoretical support for the prevention and treatment of postmenopausal osteoporosis (PMOP) using Zuogui Wan and Yougui Wan.MethodsBMSCs were divided into a blank group, Zuogui Wan (ZGW) group, Yougui Wan (YGW) group, and Progynova group. Cell identification was performed using flow cytometry. The growth curves of BMSCs were plotted using the methylthiazolyldiphenyl-tetrazolium bromide (MTT) method, and the effects of Zuogui Wan and Yougui Wan on the proliferation of BMSCs were detected. The Oil red O staining method was used to detect lipid droplet formation. The Western blot method was used to detect the expression of adipogenesis-related factors PPARγ, CCAAT/enharcer-binding protein (C/EBP)α, C/EBPβ, lipoprotein lipase (LPL) protein, brown adipose tissue-related (BAT) proteins PGC-1α, uncoupcing protein 1 (UCP1), PR domdin-containing protein 16 (PRDM16), mitochondrial biogenesis-related PGC-1α, nuclear respiratory factor 1 (Nrf1), nuclear factor E2-related factor 2 (Nrf2), and mitochondrial transcription factor A (TFAM). The expression of adipogenesis-related factors PPARγ, C/EBPα, C/EBPβ, LPL genes, and the copy number of cytochrome B (CytoB mtDNA) gene was detected using real-time polymerase chain reaction (Real-time PCR). Mitochondrial ultrastructure was detected using transmission electron microscopy.ResultsCompared with that in the blank group, the proliferation ability of BMSCs in each treatment group increased continuously as the intervention progressed, and lipid droplets significantly decreased after the drug intervention. The mRNA and protein expression levels of adipogenesis-related factors PPARγ, C/EBPα, C/EBPβ, and LPL were significantly downregulated (P<0.01), while those of the BAT-related factors PGC-1α, UCP1, PRDM16 were significantly upregulated (P<0.01). The number of mitochondria increased, accompanied by reduced swelling. The double membrane and cristae structure were clear, and the internal cristae rupture was reduced. The copy number of CytoB mtDNA in each treatment group was significantly increased (P<0.01). The protein expression levels of mitochondrial biogenesis-related PGC-1α, Nrf1, Nrf2, and TFAM in each treatment group were significantly increased (P<0.01).ConclusionBoth Zuogui Wan and Yougui Wan can prevent and treat PMOP by intervening in mitochondrial biogenesis in BMSCs through PGC-1α/PPARγ.
摘要:ObjectiveUltra performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) coupled with network pharmacology and molecular docking was utilized to explore the efficacy and mechanism of action of Ermiao Situ decoction on rats with damp-heat eczema.MethodsA rat model of damp-heat eczema was established by artificial climate chamber intervention combined with sensitization induction by dinitrochlorobenzene (DNCB), and it was randomly divided into the normal group, the model group, the medium- and high-dose groups of Ermiao Situ decoction (3.40 g·kg-1 and 6.80 g·kg-1), and the prednisone acetate group (2.51 mg·kg-1), with eight rats in each group, totalling 46 rats, of which six rats were tested with the drug-containing serum. The chemical analysis of drug-containing serum from rats was carried out by UPLC-Q-TOF-MS/MS, combined with network pharmacology for the prediction of key components, core targets, and signaling pathways, and molecular docking experiments were performed by CB-Dock2 online website. The pharmacological effects of Ermiao Situ decoction in the treatment of damp-heat eczema were investigated by epitaxial indexes combined with the pathologic tissue staining method. The serum levels of gastrin (GAS), interleukin-4 (IL-4), and interleukin-13 (IL-13) were measured by enzyme-linked immunosorbent assay (ELISA). Interleukin-6 (IL-6), Janus kinase 1 (JAK1), phosphorylated (p)-JAK1, signal transduction and activation of transcription factor 3 (STAT3), and p-STAT3 protein expression level was determined by Western bolt.ResultsA total of 19 active ingredients were detected in drug-containing serum samples of rats, which were predicted to act on 198 targets for the treatment of damp-heat eczema, among which the key ingredients included rhodopsin, huangpai alkaloids, and quercetin, and the main core targets included STAT3, tumor necrosis factor (TNF), and IL-6, which were mainly involved in the cancer signaling pathway, phosphatidylinositol 3-kinase (PI3K)/protein kinase (Akt) signaling pathway, T helper 17 (Th17) cell differentiation signaling pathway, and JAK/STAT signaling pathway. The molecular docking results suggested that the key components had strong binding activities with the core targets IL-6, JAK1, and STAT3 in the JAK/STAT signaling pathway. The results of animal experiments showed that compared with those in the normal group, rats in the model group were depressed. They had loose hair, loose stools, epidermal oozing, vesiculation, and generation of thick scabs in the form of scales, decreased body weight, increased anus temperature and water intake, and increased indexes of the spleen, thymus gland, and stomach (P<0.05, P<0.01), and the lesion tissue could be seen to be hyperkeratotic, with the aggregation of inflammatory cells and nonsignificant separation of epidermis and dermis. The gastric mucosa was thinned, deficient, and structurally disorganized, and obvious inflammatory cell aggregation was seen. The levels of GAS, IL-4, and IL-13 in serum were significantly reduced (P<0.05, P<0.01), and the protein expression levels of IL-6, JAK1, p-JAK1, and p-STAT3 in the lesion tissue were significantly increased (P<0.05, P<0.01). Compared with those in the model group, rats in each administration group had stable mental states, formed feces, a clean perianal area, and basically normal epidermis. Only a small amount of scaly scabs existed, and the rats had body weight increased, with decreased anal temperature and water intake, as well as decreased spleen, thymus, and gastric indexes (P<0.05, P<0.01). Epidermal thickness was decreased, and epidermal and dermal separation boundaries were obvious, but hyperkeratotic and accumulation of inflammatory cells could still be seen. The thickness of gastric mucosa increased, and the structure was restored to varying degrees. The levels of GAS, IL-4, and IL-13 content in the serum of rats were increased to varying degrees, and the protein expression levels of IL-6, JAK1, p-JAK1, and p-STAT3 in the dermal lesion tissue were significantly decreased (P<0.05, P<0.01).ConclusionErmiao Situ decoction may exert therapeutic effects on rats with damp-heat eczema by modulating the JAK/STAT signaling pathway.
关键词:Ermiao Situ decoction;damp-heat eczema;ultra performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS);network pharmacology;Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling pathway
摘要:ObjectiveTo investigate the improvement effect of Qibai Pingfei capsules on pulmonary vascular remodeling in rats at different stages of chronic obstructive pulmonary disease (COPD) and to analyze its possible mechanism of action.MethodsMale Sprague-Dawley (SD) rats were randomly divided into a normal group, an early COPD model group, an advanced COPD model group, an early-intervention high-dose group, a late-intervention high-dose group, an early-intervention low-dose group, a late-intervention low-dose group, an early-intervention pyrrolidine dithiocarbamate (PDTC) group, and a late-intervention PDTC group, with 15 rats in each group. A rat model of early COPD was constructed by using cigarette smoke combined with airway infusion using lipopolysaccharide(LPS), and a rat model of advanced COPD was constructed by using airway infusion with LPS, cigarette smoke, and hypoxia. All groups except the normal group were given LPS airway drops on days 1 and 14 of the experiment, smoked for 1 h per day, and administered the drug once a day for 40 weeks from day 15 onward. In the high- and low-dose groups, rats were given 1 g·kg-1 and 250 mg·kg-1 Qibai Pingfei capsules, respectively by gavage, and in PDTC groups, rats were given 100 mg·kg-1 of PDTC by intraperitoneal injection. The advanced COPD model group underwent 6 h of hypoxia per day in weeks 5-6. Lung function and mean pulmonary artery pressure were tested in rats. Morphologic changes in lung tissues were detected by hematoxylin-eosin(HE)staining. Collagen deposition in lung tissues was examined by Masson staining, and the levels of inflammatory factors including interleukin-1β(IL-1β), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α)in lung tissues were detected by enzyme-linked immunosorbent assay (ELISA). The number of inflammatory cells in the alveolar lavage fluid of rats in each group was detected by Giemsa staining, and the protein expression of Toll-like receptor 4(TLR4), myeloid differentiation factor 88(MyD88), nuclear factor-κB(NF-κB), TNF-α, vascular endothelial-cadherin(VE-cadherin), α-smooth muscle actin(α-SMA), and platelet endothelial cell adhesion molecule-1(CD31) was detected by Western blot in the lung tissues of rats.ResultsCompared with the normal group, the model group showed significantly decreased forced expiratory volume in 0.3 s (FEV0.3), forced vital capacity (FVC), and FEV0.3/FVC ratio related to lung function (P<0.05), thickening of pulmonary vasculature, increased collagen deposition in the lungs, and enhanced mean pulmonary arterial pressure and expression levels of IL-6, IL-1β, and TNF-α (P<0.05). Additionally, the model group also exhibited increased numbers of macrophages, lymphocytes, and neutrophils (P<0.05), significantly higher protein expression of TLR4, MyD88, NF-κB, TNF-α, and α-SMA (P<0.05), and significantly lower protein expression of VE-cadherin and CD31 (P<0.05). Lung function was significantly improved in the Qibai Pingfei capsules groups compared with the model group (P<0.05), with mean pulmonary arterial pressure reduced and pulmonary vascular thickening and collagen deposition in the lungs ameliorated. The Qibai Pingfei capsules groups also showed reduced expression levels of IL-6, IL-1β, and TNF-α (P<0.05) and decreased numbers of macrophages, lymphocytes, and neutrophils (P<0.05), as well as reduced protein expression of TLR4, MyD88, NF-κB, TNF-α, and α-SMA (P<0.05) and elevated protein expression of VE-cadherin and CD31 (P<0.05) in rat lung tissues.ConclusionQibai Pingfei capsules inhibits inflammatory response and endothelial-to-mesenchymal transition probably by regulating the TLR4/NF-κB signaling pathway, thus improving pulmonary vascular remodeling in COPD model rats and showing therapeutic effects in the early stage of COPD.
摘要:ObjectiveTo evaluate pharmacological effects of Shengmai injection(SMI)on cerebral ischemia and study its neuroprotective mechanism.MethodsMale specific pathogen-free (SPF) Sprague-Dawley (SD) rats were randomly divided into a sham group, a model group, a low-dose SMI group(3 mL·kg-1), a middle-dose SMI group(6 mL·kg-1), a high-dose SMI group(12 mL·kg-1), and a Ginaton group(4 mL·kg-1)according to the random number table method, with 12 rats in each group. The rat model of cerebral ischemia-reperfusion(MCAO/R)was prepared via the suture method. The administration groups were intraperitoneally injected with corresponding concentrations of SMI or Ginaton injection after reperfusion, which was conducted for 3 consecutive days. The sham group and model group were administered the equivalent volume of physiological saline. The pharmacological effects of SMI on brain injury in MCAO/R rats were evaluated by neurological function scores, cerebral infarction area, hematoxylin-eosin (HE) staining, Nissl staining, terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining, and Western blot. The dominant link and key protein of SMI treating cerebral injury were explored using proteomic analysis. The related mechanisms of SMI were further validated using enzyme-linked immunosorbent assay (ELISA), Western blot, and chloride ion fluorescence probe with oxygen-glucose deprivation/reoxygenation(OGD/R)-treated PC12 cells and MCAO/R rats.ResultsCompared with the sham group, the model group showed significantly increased neurological function scores, cerebral infarction area, neuronal apoptosis rate, and expression levels of apoptosis related proteins (P<0.05, P<0.01)and significantly decreased density of Nissl bodies and neurons(P<0.01). Compared with the model group, the SMI groups exhibited significantly decreased neurological function scores, cerebral infarction area, neuronal apoptosis rate, and expression levels of apoptosis related proteins (P<0.05, P<0.01)and significantly increased density of Nissl bodies and neurons (P<0.05). The proteomic analysis results showed that oxidative stress and inflammatory response were important processes of SMI intervening in MCAO/R injury, and the chloride intracellular channel protein 1 (CLIC1) was one of key proteins in its action network. The levels of representative indicators of oxidative stress and inflammatory response in the MCAO/R rats of the SMI groups were significantly reduced, compared with those in the model group(P<0.05, P<0.01), and the expression levels of CLIC1 and downstream NOD-like receptor protein 3 (NLRP3) decreased (P<0.01). In addition, the experimental results based on the OGD/R PC12 cells showed that SMI significantly increased the cell survival rate(P<0.01) and significantly decreased the intracellular chloride ion concentration(P<0.05).ConclusionSMI has neuroprotective effects. Oxidative stress and inflammatory response are key processes of SMI intervening in MCAO/R injury. The potential mechanism is closely related to the regulation of CLIC1.
WANG Tengjian, ZHAO Wanlu, YU Yang, LIU Yan, CAO Kun, LIN Zheyuan, WU Yue, LUO Lilan, LAI Weizhi, LOU Zhaohuan, ZHANG Qiaoyan, ZHANG Quanlong, QIN Luping
摘要:ObjectiveTo investigate the optimal ratio of goat horn replacing antelope horn in Fufang Lingjiao Jiangya pills and the blood pressure-lowering mechanism of this medicine.MethodsThe blood pressure-lowering efficacy of Fufang Lingjiao Jiangya pills with varying proportions of goat horn replacing antelope horn was evaluated on spontaneous hypertensive rats (SHR). In this experiment, 50 SHR rats were randomly grouped as follows: model (n=8), captopril (0.01 g·kg-1) (n=6), low-dose blank Fufang Lingjiao Jiangya pills (0.342 g·kg-1) (n=6), high-dose blank Fufang Lingjiao Jiangya pills (0.684 g·kg-1) (n=6), low-dose antelope horn-containing Fufang Lingjiao Jiangya pills (0.378 g·kg-1) (n=6), high-dose antelope horn-containing Fufang Lingjiao Jiangya pills (0.756 g·kg-1) (n=6), low-dose goat horn-containing Fufang Lingjiao Jiangya pills (0.378 g·kg-1) (n=6), and high-dose goat horn-containing Fufang Lingjiao Jiangya pills (0.756 g·kg-1) (n=6). Additionally, 8 WKY rats were used as the normal group. Drugs were administered by gavage for 4 weeks while an equal volume of distilled water was administered for the normal and model groups. Blood pressure was measured before administration, 3 h post administration, and biweekly thereafter. In the experiment for Fufang Lingjiao Jiangya pills with goat horn replacing antelope horn in different proportions, 48 SHR rats were randomly grouped as follows: model, blank Fufang Lingjiao Jiangya pills (0.684 g·kg-1), antelope horn-containing Fufang Lingjiao Jiangya pills (0.756 g·kg-1), 2× goat horn-containing Fufang Lingjiao Jiangya pills (0.824 g·kg-1), 4× goat horn Fufang Lingjiao Jiangya pills (0.969 g·kg-1), and 6× goat horn Fufang Lingjiao Jiangya pills (1.112 g·kg-1). The normal group included 8 WKY rats, and the normal group and model group received an equal volume of distilled water. The treatment lasted for 2 weeks, and blood pressure was recorded at various time points (pre-administration, 3 h post administration, and on days 4, 7, 10, and 14 of administration). Serum levels of angiotensin-converting enzyme (ACE), angiotensin Ⅱ(Ang Ⅱ), renin, and interleukin-6 (IL-6) were measured by enzyme-linked immunosorbent assay. Histopathological changes in the heart, kidney, and thoracic aorta were observed by hematoxylin-eosin staining. The protein levels of ACE2, angiotensin Ⅱ type 1 receptor (AT1R), and angiotensinogen (AGT) in the kidney tissue were determined by Western blot, while the expression of nuclear factor (NF)-κB p65 and Toll-like receptor 4 (TLR4) in the thoracic aorta tissue was assessed by immunohistochemistry.ResultsCompared with the model group, all treatment groups showed lowered blood pressure (P<0.05, P<0.01), and the 6× goat horn-containing Fufang Lingjiao Jiangya pills group showed consistent blood pressure-lowering effect with the antelope horn-containing Fufang Lingjiao Jiangya pills group. Compared with the normal group, the model group showed elevated serum levels of ACE, Ang Ⅱ, renin, and IL-6, while the elevations were declined in the Fufang Lingjiao Jiangya pills groups (P<0.05, P<0.01). Pathological changes in the heart, kidney, and thoracic aorta were alleviated in all the treatment groups, with the 6× goat horn- and antelope horn-containing Fufang Lingjiao Jiangya pills groups exhibited the best effect. Western blot and immunohistochemistry results showed that all the treatment groups exhibited down-regulated protein levels of AT1R, AGT, NF-κB p65, and TLR4 and up-regulated protein levels of ACE2 (P<0.05, P<0.01) compared with model group, with the 6×goat horn- and antelope horn-containing Fufang Lingjiao Jiangya pills groups showcasing the best effect.ConclusionReplacing antelope horn with 6×goat horn in Fufang Lingjiao Jiangya pills can achieve consistent blood pressure-lowering effect with the original prescription. The prescription may exert the effect by inhibiting the renin-angiotensin-aldosterone system (RAAS) and TLR4/NF-κB signaling pathways.
摘要:ObjectiveTo investigate the mechanisms through which Shugan Bushen Yulin decoction ameliorates spermatogenic dysfunction and sperm quality degradation caused by propylthiouracil (PTU) via the endoplasmic reticulum stress pathway in rats.MethodsSixty male rats were randomly assigned into six groups: a control group, a model group, low- (6.75 g·kg-1), medium- (13.5 g·kg-1), and high-dose (27 g·kg-1) Shugan Bushen Yulin decoction groups, and an L-carnitine (0.27 g·kg-1) group, with ten rats in each group. Other groups except the control group were administrated with PTU (10 mg·kg-1) by gavage for 12 consecutive days. After the completion of modeling, rats were administrated with corresponding agents or normal saline (control group) via gavage for 28 consecutive days. Twenty-four hours after the last administration, rats were sacrificed, and the body and organ (testis, epididymis, and seminal vesicle) weights were measured to calculate the organ indices. Hematoxylin-eosin staining was employed to observe the pathological changes in the testes and epididymis, and the testicular spermatogenic function of rats was scored. Enzyme-linked immunosorbent assay was employed to measure the levels of thyroid-stimulating hormone (TSH), triiodothyronine (T3), thyroxine (T4), follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), estradiol (E2), and malondialdehyde (MDA) and the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the rat serum. Terminal deoxynucleotidyl transferase dUTP nick end labeling was adopted to assess the rate of testicular cell apoptosis. Western blot was conducted to determine the expression levels of glucose-regulated protein 78 (GRP78), CCAAT/enhancer-binding protein homologous protein (CHOP), phosphorylated protein kinase RNA-like endoplasmic reticulum kinase (p-PERK), protein kinase RNA-like endoplasmic reticulum kinase (PERK), phosphorylated eukaryotic translation initiation factor 2α kinase (p-EIF2α), eukaryotic translation initiation factor 2α kinase (EIF2α), and activating transcription factor 4 (ATF4) in the testicular tissue of rats.ResultsCompared with the control group, the model group exhibited reductions in volumes and indexes of testes, epididymides, and seminal vesicles (P<0.01), pathological damage of testes and epididymides, and declines in spermatogenic function and sperm density and motility (P<0.01). In addition, the model group demonstrated elevated serum levels of TSH and MDA, lowered levels of T3, T4, FSH, LH, T, and E2, decreased activities of SOD and GSH-Px (P<0.05, P<0.01), an increased apoptosis rate of testicular cells (P<0.01), and up-regulated expression of GRP78, CHOP, p-PERK/PERK, p-EIF2α/EIF2α, and ATF4 in the testicular tissue (P<0.01). Compared with the model group, Shugan Bushen Yulin decoction at different doses improved the morphology and indexes of testes, epididymides, and seminal vesicles (P<0.05), restored the spermatogenic function (P<0.05, P<0.01), increased the sperm density and motility (P<0.05, P<0.01), and decreased the percentage of apoptotic testicular cells (P<0.05). Furthermore, the decoction lowered the serum level of MDA, elevated the serum levels of FSH, LH, T, and E2, and increased the activities of SOD and GSH-Px (P<0.05, P<0.01). Moreover, the expression levels of GRP78, CHOP, p-PERK/PERK, p-EIF2α/EIF2α, and ATF4 in the testicular tissue were down-regulated (P<0.05, P<0.01).ConclusionShugan Bushen Yulin decoction can ameliorate the spermatogenic dysfunction and sperm quality degradation induced by PTU in a rat model of hypothyroidism by repairing the testicular oxidative damage and regulating the PERK/EIF2α/ATF4 signaling pathway, thereby alleviating spermatogenic cell apoptosis.
摘要:ObjectiveTo investigate the effect of Qingrun prescription(QRP)-containing serum on improving insulin resistance in HepG2 cells and its potential mechanisms.MethodsAn insulin resistance model was established in HepG2 cells with 1×10-6 mol·L-1 insulin. Branched-chain α-keto acid dehydrogenase (BCKDH) gene silencing was achieved using siRNA, and the cells were divided into 8 groups: normal group, model group (1×10-6 mol·L-1 insulin), metformin group (1 mmol·L-1 metformin), high-, medium-, and low-dose QRP groups (20%, 10%, and 5% QRP-containing serum, respectively), QRP + siRNA-silenced BCKDH (si-BCKDH) group (10% QRP-containing serum + si-BCKDH), and QRP + si-NC group (10% QRP-containing serum + si-NC). Glucose levels in the supernatant were measured with a glucose assay kit, while glycogen content was assessed using a glycogen assay kit. Levels of branched-chain amino acids (BCAAs) and branched-chain keto acids (BCKAs) were determined using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). mRNA transcription and protein expression levels of BCKDH, dishevelled, Egl-10, and pleckstrin (DEP) domain-containing mammalian target of rapamycin (mTOR)-interacting protein (DEPTOR), mTOR, and ribosomal protein S6 kinase 1 (S6K1) were detected using real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot.ResultsCompared to the normal group, the model group exhibited significantly decreased glucose consumption and glycogen content, increased levels of BCAAs and BCKAs, downregulated expression of BCKDH and DEPTOR, and upregulated mTOR and S6K1 expression (P<0.01). In comparison to the model group, QRP treatment at all doses significantly enhanced glucose consumption and glycogen content while reducing BCAAs and BCKAs levels (P<0.01). The high- and medium-dose QRP groups demonstrated significant upregulation of BCKDH mRNA transcription and protein expression, as well as DEPTOR mRNA transcription. Moreover, the DEPTOR protein expression level was significantly increased in high-, medium-, and low-dose QRP groups, while mTOR and S6K1 mRNA and protein expression levels were markedly downregulated (P<0.05, P<0.01). Compared to the QRP + si-NC group, the QRP + si-BCKDH group exhibited increased BCAAs and BCKAs levels, significantly decreased BCKDH mRNA transcription and protein expression, downregulated DEPTOR mRNA and protein expression, and upregulated mTOR and S6K1 mRNA and protein expression (P<0.05, P<0.01).ConclusionQRP may improve insulin resistance by reprogramming BCAAs metabolism. This effect involves upregulating BCKDH, reducing BCAAs and BCKAs levels, and suppressing the mTOR pathway activation.
摘要:ObjectiveTo explore the role and mechanism of Hei Xiaoyaosan in regulating the protein kinase B (Akt)/glycogen synthase kinase-3β (GSK-3β)/nuclear factor E2-related factor 2 (Nrf2)/heme oxygenase-1 (HO-1) signaling pathway in cascade modulation of oxidative stress and apoptosis for preventing and treating Alzheimer's disease (AD).MethodsNinety male SD rats of 4 months old were randomly assigned into a control group (n=10), a sham group (with injection of 1 μL normal saline into bilateral hippocampi, n=10), and a modeling group (with injection of 1 μL beta-amyloid 1-42 solution into bilateral hippocampi to induce AD, n=70). One week after modeling, 50 successfully modeled rats were selected and randomly allocated into model, donepezil hydrochloride (0.45 mg·kg-1), and high-, medium-, and low-dose (15.30, 7.65, 3.82 g·kg-1, respectively) Hei Xiaoyaosan groups. The rats were administrated with corresponding drugs once daily for six consecutive weeks. The Morris water maze was used to assess the learning and memory abilities of rats. Hematoxylin-eosin (HE) staining was performed to reveal hippocampal morphological changes in AD rats. Apoptosis in the hippocampal CA3 region was detected by terminal-deoxynucleotidyl transferase-mediated Nick end labeling. Immunofluorescence was used to visualize the expression of neuronal nuclear antigen (NeuN) in the CA1 region. Additionally, enzyme-linked immunosorbent assay was performed to assess the activities of glutathione peroxidase (GSH-Px), glutathione-S-transferase (GST), and catalase (CAT) in the hippocampus. Real-time PCR was conducted to measure the mRNA levels of Akt, GSK-3β, Nrf2, and HO-1, while Western blot was employed to determine the protein levels of phosphorylated Akt (p-Akt)/Akt, phosphorylated GSK-3β (p-GSK-3β)/GSK-3β, Nrf2, and HO-1.ResultsCompared with the control group, the model group on day 5 showed an increase in total swimming distance (P<0.01), a reduction in the percentage of time spent in the target quadrant (P<0.01), reduced and disarranged neurons, nuclear condensation, varying degrees of cellular damage, increased apoptosis of hippocampal neurons (P<0.01), decreased NeuN content (P<0.01), weakend activities of GSH-Px, GST, and CAT (P<0.01), and down-regulated mRNA levels of Nrf2 and HO-1 (P<0.01) and protein levels of p-Akt/Akt, p-GSK-3β/GSK-3β, Nrf2, and HO-1 (P<0.01) in the hippocampus. Compared with the model group, donepezil hydrochloride and high, medium, and low doses of Hei Xiaoyaosan shortened the total swimming distance on day 5 (P<0.05, P<0.01), increased the percentage of time spent in the target quadrant (P<0.05, P<0.01), improved the arrangement and morphology of neurons, reduced nuclear condensation and the apoptosis rate of hippocampal neurons (P<0.01), increased the NeuN content (P<0.01), enhanced the activities of GSH-Px, GST, and CAT (P<0.05, P<0.01), and up-regulated the mRNA levels of Nrf2 and HO-1 (P<0.05, P<0.01) and the protein levels of p-Akt/Akt, p-GSK-3β/GSK-3β, Nrf2, and HO-1 (P<0.05, P<0.01) in the hippocampus.ConclusionHei Xiaoyaosan can regulate the Akt/GSK-3β/Nrf2/HO-1 pathway to enhance the antioxidant stress capacity and inhibit neuron apoptosis to exert the neuroprotective effect, thereby ameliorating the cognitive dysfunction and pathological damage in AD rats.
摘要:ObjectiveTo explore the mechanism of Hei Xiaoyaosan in improving the cognitive function in Alzheimer's disease (AD) from cell apoptosis mediated by the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/nuclear factor kappa B (NF-κB) signaling pathway.MethodsFour-month-old SD male rats were randomly assigned into a blank group, a sham group, a model group, a donepezil hydrochloride (0.45 mg·kg-1) group, and high-, medium-, and low-dose (15.30, 7.65, and 3.82 g·kg-1, respectively) Hei Xiaoyaosan groups, with 10 rats in each group. The sham group received bilateral hippocampal injection of 1 μL normal saline, while the other groups received bilateral hippocampal injection of 1 μL beta-amyloid 1-42 (Aβ1-42) solution for the modeling of AD. Rats were administrated with corresponding agents once a day for 42 consecutive days. The Morris water maze test was carried out to assess the learning and memory abilities of rats. Hematoxylin-eosin staining was employed to observe pathological changes in the hippocampus of rats. Enzyme-linked immunosorbent assay was employed to measure the levels of cysteinyl aspartate-specific proteinase-3 (Caspase-3), B-cell lymphoma-2 (Bcl-2), and Bcl-2-associated X protein (Bax). Western blot was employed to determine the protein levels of PI3K, Akt, and NF-κB. A cell model of AD was established by co-culturing Aβ1-42 and PC12 cells in vitro. Cell viability and apoptosis were detected by the cell-counting kit 8 (CCK-8) assay and flow cytometry (FC), respectively.ResultsAnimal experiments showed that compared with the blank group, the model group had a prolonged escape latency (P<0.01), a reduced number of crossing platforms (P<0.01), disarrangement and a reduced number of hippocampal neurons, up-regulated expression of Bax and Caspase-3, down-regulated expression of Bcl-2 (P<0.01), decreased p-PI3K/PI3K and p-Akt/Akt levels, and an increased p-NF-κB/NF-κB level (P<0.01). Compared with the model group, donepezil hydrochloride and high- and medium-dose Hei Xiaoyaosan shortened the escape latency and increased the number of crossing platforms (P<0.05, P<0.01), improved the arrangement and increased the number of hippocampal neurons, down-regulated the expression levels of Bax and Caspase-3, up-reguated the expression level of Bcl-2 (P<0.05, P<0.01), increased the p-PI3K/PI3K and p-Akt/Akt levels (P<0.05, P<0.01), and reduced the p-NF-κB/NF-κB level (P<0.05, P<0.01). Cell experiments showed that compared with the blank group, the model group exhibited an increased apoptosis rate (P<0.01). Compared with the model group, the serum containing Hei Xiaoyaosan at various doses improved the cell viability (P<0.01), and the serum containing Hei Xiaoyaosan at the high dose decreased the cell apoptosis (P<0.01).ConclusionHei Xiaoyaosan may improve the learning and memory abilities of AD model rats by regulating cell apoptosis, while increasing the vitality and reducing the apoptosis rate of AD model cells via the PI3K/Akt/NF-κB signaling pathway.
关键词:Alzheimer's disease;Hei Xiaoyaosan;cell apoptosis;phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/nuclear factor-kappa B (NF-κB) signaling pathway
摘要:ObjectiveTo investigate the effects of Hei Xiaoyaosan on cognitive impairment and the histone deacetylase sirtuin-1 (SIRT1)/tumor suppressor p53/solute carrier family 7 member 11 (SLC7A11) signaling pathway in the rat model of Alzheimer's disease (AD).MethodsA total of 90 16-week-old SPF-grade SD male rats were randomly assigned in a blank group (n=10), a sham group (n=10, with injection of 1 μL normal saline into the bilateral hippocampi), and an AD modeling group (n=70, with injection of 1 μL β amyloid 1-42 solution into the bilateral hippocampi). According to the random number table method, fifty successfully modeled rats were assigned into model, donepezil hydrochloride (0.45 mg·kg-1), and high-, medium-, and low-dose (15.30, 7.65, and 3.82 g·kg-1, respectively) Hei Xiaoyaosan groups, and they were administrated with corresponding agents via gavage once a day for 42 consecutive days. Morris water maze test was carried out to examine the cognitive function of rats. Nissl staining was employed to observe the morphology of hippocampal neurons in each group, and Prussian blue staining was used to detect iron deposition in the hippocampal tissue. Biochemical kits were used to measure the levels of superoxide dismutase (SOD), malondialdehyde (MDA), and iron ion (Fe2+) in the hippocampal tissue. Western blot and real-time quantitative polymerase chain reaction (Real-time PCR) were employed to determine the protein and mRNA levels, respectively, of SIRT1, p53, SLC7A11, glutathione peroxidase 4 (GPX4), and acyl-CoA synthetase long-chain family member 4 (ACSL4) in the hippocampus.ResultsCompared with the blank group, the model group showed reductions in target quadrant movement distance (P<0.01) and target quadrant residence time (P<0.05), disarrangement of hippocampal neurons, increased ferroptosis deposition in the hippocampus, a lowered level of SOD, risen levels of MDA and Fe2+ (P<0.05, P<0.01), down-regulated protein and mRNA levels of SIRT1, SLC7A11, and GPX4 (P<0.05, P<0.01), up-regulated protein and mRNA levels of p53 and ACSL4 (P<0.01), and aggravated pathological process of AD. Compared with the model group, donepezil hydrochloride extended the target quadrant residence time and the target quadrant movement distance (P<0.05, P<0.01). High- and medium- doses of Hei Xiaoyaosan extended the target quadrant residence time and the target quadrant movement distance (P<0.05, P<0.01), improved the neuron arrangement and reduced the ferroptosis deposition in the hippocampus, elevated the SOD level, lowered the MDA and Fe2+ levels (P<0.05, P<0.01), up-regulated the protein and mRNA levels of SIRT1, SLC7A11, and GPX4 (P<0.01, P<0.05), and down-regulated the protein and mRNA levels of p53 and ACSL4 (P<0.05, P<0.01).ConclusionHei Xiaoyaosan can regulate the SIRT1/p53/SLC7A11 signaling pathway to mitigate oxidative stress and inhibit ferroptosis, thereby ameliorating the cognitive dysfunction in AD rats.
关键词:Alzheimer's disease;Hei Xiaoyaosan;histone deacetylase sirtuin-1 (SIRT1)/tumor suppressor p53/solute carrier family 7 member 11 (SLC7A11) signaling pathway;ferroptosiss
摘要:ObjectiveTo observe the effects of Hei Xiaoyaosan on the learning and memory abilities of Alzheimer's disease model mice (APP/PS1 mice), and to explore its mechanism through the inflammatory cascade mediated by nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3)/cysteine aspartate-specific protease (Caspase-1)/gasdermin D (GSDMD) signaling pathway.MethodsSPF-grade 4-month-old APP/PS1 mice were randomly divided into the model group, MCC950 group, and Hei Xiaoyaosan high-, medium-, and low-dose groups. C57BL/6J mice were used as the blank group. After 7 days of adaptive feeding, mice in each group were intervened. The Hei Xiaoyaosan high-, medium-, and low-dose groups were given corresponding doses by gavage (25.79, 12.90, 6.45 g·kg-1·d-1), the MCC950 group was intraperitoneally injected with 10 mg·kg-1·2 d-1, and the blank group received the same volume of physiological saline by gavage. After 90 days of intervention, the learning and memory abilities were assessed using the Y maze and Morris water maze tests. The structural changes of hippocampal neurons were observed by hematoxylin-eosin (HE) staining. The expression of amyloid precursor protein (APP) in the hippocampal CA3 region was detected by immunohistochemistry. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of interleukin (IL)-10, IL-18, and IL-1β in the hippocampus. Western blot was applied to detect the protein expression of NLRP3, Caspase-1, GSDMD, and GSDMD-N in the hippocampus. Immunofluorescence was used to detect the co-localization of GSDMD-N and ionized calcium-binding adapter molecule-1 (Iba-1) in the hippocampus.Results① In the Y maze test, compared with the blank group, the spontaneous alternation rate of the model group was significantly reduced (P<0.01). Compared with the model group, the spontaneous alternation rate in the Hei Xiaoyaosan high- and low-dose groups was significantly increased (P<0.01). ② In the Morris water maze test, during the 1-4 days of the location navigation test, the escape latency time of mice decreased with the extension of training time. On day 4, compared with the blank group, the model group showed a significantly increased escape latency (P<0.05). Compared with the model group, the MCC950 group and the Hei Xiaoyaosan low-dose group showed significantly reduced escape latency (P<0.05). In the spatial exploration experiment, compared with the blank group, the number of platform crossings in the model group was significantly reduced (P<0.01). Compared with the model group, the Hei Xiaoyaosan low-dose group showed significantly increased platform crossings (P<0.05). ③ HE staining showed that, compared with the blank group, the hippocampal CA3 cells of the model group were damaged, arranged loosely and irregularly, swollen, with unclear boundaries, and the nuclei were pyknotic and deeply stained. MCC950 and all doses of Hei Xiaoyaosan improved the hippocampal CA3 cell damage in APP/PS1 mice to varying degrees. ④ Immunohistochemical results indicated that, compared with the blank group, the expression of APP in the hippocampal CA3 region was significantly increased in the model group (P<0.01). MCC950 and all doses of Hei Xiaoyaosan could reduce the expression of APP in the hippocampal CA3 region of APP/PS1 mice (P<0.01). ⑤ ELISA results showed that the levels of IL-18 and IL-1β in the hippocampus of mice in the model group were significantly increased, and IL-10 levels were significantly reduced (P<0.01). Compared with the model group, the IL-18 levels in the MCC950 group and the Hei Xiaoyaosan medium- and low-dose groups were significantly reduced (P<0.01). IL-1β levels in the hippocampus of the MCC950 group and Hei Xiaoyaosan high-, medium-, and low-dose groups were significantly decreased (P<0.01). The IL-10 levels in the hippocampus of the MCC950 group and the Hei Xiaoyaosan medium- and low-dose groups were increased (P<0.05, P<0.01). ⑥ Western blot results showed that compared with the blank group, the protein levels of NLRP3, Caspase-1, GSDMD, and GSDMD-N in the hippocampus of the model group were significantly elevated (P<0.01). Compared with the model group, the content of NLRP3 and Caspase-1 in the hippocampus of the treated groups was decreased (P<0.05, P<0.01). The content of GSDMD in the hippocampus of the Hei Xiaoyaosan high-, medium-, and low-dose groups was reduced (P<0.05, P<0.01), and the content of GSDMD-N in the hippocampus of the Hei Xiaoyaosan medium- and low-dose groups was decreased (P<0.05, P<0.01). ⑦ Immunofluorescence results showed that, compared with the blank group, the co-expression of GSDMD-N and Iba-1 in the hippocampus of the model group was significantly increased (P<0.01). Compared with the model group, the co-expression of GSDMD-N and Iba-1 in the treated groups was significantly reduced (P<0.01).ConclusionHei Xiaoyaosan may regulate the NLRP3/Caspase-1/GSDMD signaling pathway to affect the release of inflammatory factors, alleviate neuroinflammation,improve hippocampal histopathological changes,and improve learning and memory deficits,thus providing potential therapeutic benefits for Alzheimer's disease.
关键词:Alzheimer's disease;Hei Xiaoyaosan;neuroinflammation;nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3(NLRP3)/cysteine aspartate-specific protease(Caspase-1)/gasdermin D(GSDMD) signaling pathways;experimental study
摘要:ObjectiveTo establish preliminary diagnostic criteria for spleen and kidney deficiency syndrome in ischemic stroke patients and provide a basis for standardized diagnosis and treatment of ischemic stroke.MethodsRelevant literature on the diagnostic criteria for spleen and kidney deficiency syndrome in ischemic stroke patients was retrieved, and data were mined and extracted to form an item pool. Based on the formation of the item pool, this study used the Delphi method to initiate two rounds of questionnaire surveys with selected experts to complete the initial screening of items and the discrimination of symptom importance. A prospective clinical investigation method was adopted to collect clinical information from patients, and statistical analysis methods and data mining techniques were comprehensively used to determine their primary and secondary symptoms. Based on the clear main and secondary symptoms identified, combined with expert group discussions, the study established preliminary diagnostic criteria for spleen and kidney deficiency syndrome in ischemic stroke patients.ResultsA total of 25 relevant syndrome differentiation standards were included. After splitting, standardizing, and screening the items, the study established a pool of 48 items. The first round of questionnaire survey consulted 30 experts, with both the positive coefficient of experts and the effectiveness rate of the questionnaire reaching 100%. The Kendall's coefficient of concordance was 0.359. According to the item screening criteria, 26 items were retained in this round of questionnaire survey. A total of 176 cases were collected through clinical information investigation, including 94 cases with spleen and kidney deficiency syndrome and 82 cases without spleen and kidney deficiency syndrome. The statistical results were as follows: ① Descriptive statistics: The main symptoms with a frequency of ≥ 30% included mental fatigue and lack of strength, weakness of the lower back and knees, etc. The secondary symptoms with a frequency of ≥ 10% and ≤ 30% were lassitude and disinclination to talk, shortness of breath, etc. ② Binary logistic regression analysis: The main symptoms with an odds ratio (OR) value of ≥ 3 were mental fatigue and lack of strength, weakness of the lower back and knees, etc. The secondary symptoms with an OR value of ≥ 1 and ≤ 3 were lassitude and disinclination to talk, shortness of breath, etc. Artificial neural network: The main symptoms with a weight value(Wij)of ≥ 0.5 and < 1 were mental fatigue and lack of strength, lassitude and disinclination to talk, etc. The secondary symptoms with Wij of ≥ 0.3 and < 0.5 were shortness of breath, flaccid limbs, etc. In the second round of questionnaire survey, a total of 37 experts were consulted, with both the positive coefficient of experts and the effectiveness rate of the questionnaire reaching 100%. The Kendall's coefficient of concordance was 0.237. According to the criteria to determine primary and secondary symptoms based on the Delphi method, the main symptoms included in this round of the questionnaire were mental fatigue and lack of strength, lassitude and disinclination to talk, etc., and the secondary symptoms were shortness of breath, dizziness, etc.ConclusionThe main symptoms of spleen and kidney deficiency syndrome in ischemic stroke patients are mental fatigue and lack of strength, weakness of the lower back and knees, loose stool, pale and edematous tongue texture possibly with tooth marks, and deep and thready pulse or weak pulse. The secondary symptoms include shortness of breath, dizziness, tinnitus and deafness, decreased appetite or postprandial abdominal distension, pale complexion, frequent micturition at night, dull tongue texture, and white and slippery tongue coating. The preliminarily established diagnostic criteria for spleen and kidney deficiency syndrome in ischemic stroke patients can provide a standardized and objective basis, thereby better guiding clinical diagnosis and treatment of ischemic stroke.
关键词:ischemic stroke;spleen and kidney deficiency syndrome;Delphi method;expert questionnaire;clinical investigation;diagnostic criteria
摘要:ObjectiveTo observe the clinical effect of modified Huanglian Wendantang on cardiovascular risk factors in patients with metabolic syndrome under the guidance of treating disease before its onset.MethodsA total of 82 patients with metabolic syndrome treated in the First Affiliated Hospital of Heilongjiang University of Chinese Medicine from July 2023 to July 2024 were selected and allocated into an observation group (41 cases) and a control group (41 cases) by the random number table method. The control group received routine treatment, and the observation group was treated with modified Huanglian Wendantang on the basis of routine treatment. Both groups were treated for 8 weeks. The therapeutic effects on TCM symptoms after treatment in the two groups were evaluated. The levels of obesity degree indicators, blood pressure indicators, glucose and lipid metabolism indicators, inflammatory factors, and vascular endothelial function indicators before and after treatment in the two groups were measured, and the treatment safety was evaluated.ResultsAfter treatment, the total response rate of TCM symptoms in the observation group was 97.56% (40/41), which was higher than that (87.80%, 36/41) in the control group (χ2=5.205, P<0.05). After treatment, both groups showed declines (P<0.05) in systolic blood pressure (SBD), diastolic blood pressure (DBP), triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), fasting blood glucose, 2-hour postprandial blood glucose (2 h PG), glycosylated hemoglobin (HbA1c), fasting insulin (FINS), Homeostatic Model Assessment for Insulin Resistance (HOMA-IR), body mass index (BMI), waist circumference (WC), waist-to-hip ratio (WHR), leptin (LEP), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), endothelin-1 (ET-1), and inducible nitric oxide synthase (iNOS). Moreover, the declines in the observation group were more obvious than those in the control group (P<0.05, P<0.01). After treatment, both groups showed elevated levels of high density lipoprotein cholesterol (HDL-C), adiponectin (ADP), nitric oxide (NO), and endothelial nitric oxide synthase (eNOS) (P<0.05), and the above indexes in the observation group were higher than those in the control group (P<0.01).ConclusionUnder the guidance of the thought of treating disease before its onset, modified Huanglian Wendantang was used to treat patients with metabolic syndrome. The decoction improved the clinical efficacy by ameliorating IR to improve insulin sensitivity, reducing inflammation, and protecting the vascular endothelial function. It inhibits cardiovascular risk factors without inducing adverse reactions, being worthy of clinical application and promotion.
关键词:treating disease before its onset;metabolic syndrome;Huanglian Wendantang;cardiovascular risk factors;clinical observation
摘要:ObjectiveTo observe the clinical efficacy of Tongfu Kuanzhong decoction (TFKZ) in the treatment of constipation-predominant irritable bowel syndrome (IBS-C) with spleen deficiency and Qi stagnation and its effects on anorectal manometry and colonic electrical activity parameters.MethodsSixty-four patients with IBS-C of the spleen deficiency and Qi stagnation type were included as research subjects and were randomized into the observation group (thirty-two cases) and control group (thirty-two cases). The control group was orally administered mosapride citrate tablets, and the observation group was orally administered TFKZ. Both groups were treated for 4 weeks. The traditional Chinese medicine (TCM) syndrome scores, patient assessment of constipation symptoms (PAC-SYM) score, general anxiety disorder-7 (GAD-7) score, patient health questionnaire-9(PHQ-9)score, anorectal manometry, and colonic electrical activity parameters were observed before and after treatment in the two groups.ResultsThe total points of TCM syndromes were significantly lower in both groups after treatment (P<0.01) and lower in the observation group (P<0.05). The fecal symptoms, rectal symptoms, abdominal symptoms, and total symptom scores of the patients in both groups were significantly decreased after treatment (P<0.01), and the rectal symptoms, abdominal symptoms, and total symptom scores of the observation group were lower (P<0.05,P<0.01). In both groups, the forced anal residual pressure and the initial rectal sensation threshold decreased after treatment (P<0.05,P<0.01), and forced anal residual pressure in the control group was lower (P<0.01). After treatment, the GAD-7 and PHQ-9 scores of patients in both groups decreased significantly (P<0.01), and the GAD-7 score of the observation group was significantly lower (P<0.01). After treatment, the average wave amplitude at different time points of each lead increased significantly in both groups, compared with that before treatment (P<0.01). The average wave amplitude of each lead of the ascending colon, descending colon, sigmoid colon, and rectum in the observation group at 5 min before the meal and 10 min after the meal, as well as that of the ascending colon and rectum at 20 min and 30 min after the meal, was higher than that of the control group (P<0.05).ConclusionTFKZ is effective in the treatment of IBS-C. Its mechanism may be related to the regulation of colonic function by down-regulating the forced anal residual pressure and the initial rectal sensation threshold and up-regulating the amplitude and frequency of intestinal electrical activity in each intestinal segment of the colon.
摘要:ObjectiveTo evaluate the clinical effects of Yishen Tongluo Prescription in treating patients with idiopathic asthenozoospermia and teratozoospermia due to kidney deficiency and collateral obstruction, as well as the effects of this formula on biochemical indices and oxidative stress of seminal plasma.MethodsPatients who fulfilled the inclusion criteria were randomly allocated by the random number table method into an observation group (61 patients, including 2 dropped out and 59 completed) and a control group (61 patients, including 2 dropped out, 1 excluded, and 58 completed). The observation group took Yishen Tongluo Prescription orally, 1 dose/day, and the control group took levocarnitine oral solution, 10 mL/time, 3 times/day. Both groups received a treatment course of 12 weeks and were followed up for 6 weeks. The percentage of progressive motile sperms (PR), percentage of sperms with normal morphology, seminal plasma oxidative stress indicators [malondialdehyde (MDA) and superoxide dismutase (SOD)], seminal plasma biochemical indicators [acid phosphatase (ACP), zinc (Zn), fructose (Fru), and alpha-glucosidase (α-Glu)], and traditional Chinese medicine (TCM) symptom scores were recorded for both groups before and after treatment.ResultsThe total response rate in the observation group was 76.3% (45/59), which was higher than that (44.8%, 26/58) in the control group (χ2=12.120, P<0.05). After treatment, both groups of patients showed increases in PR, percentage of sperms with normal morphology, and seminal plasma SOD level (P<0.05), which were higher in the observation group than in the control group (P<0.05). The MDA level and TCM symptom scores decreased in both groups after treatment (P<0.05), and the decreases were more obvious in the observation group than in the control group (P<0.05). After treatment, the seminal plasma levels of ACP, Zn, Fru, and α-Glu in the observation group (P<0.05) and that of α-Glu in the control group (P<0.05) rose. Moreover, the observation group had higher seminal plasma levels of ACP, Zn, Fru, and α-Glu than the control group (P<0.05). Neither group showed obvious adverse reactions.ConclusionIn summaryYishen Tongluo prescription is able to improve the sperm vitality and reduce the sperm abnormality rate in infertile patients with idiopathic asthenozoospermia and teratozoospermia due to kidney deficiency and collateral obstruction. It may exert therapeutic effects by regulating the microenvironments of the epididymis, prostate, and seminal vesicles, thereby recovering the seminal plasma biochemical levels, enhancing the antioxidant capacity of seminal plasma, clearing excessive reactive oxygen species, and alleviating oxidative stress.
关键词:Idiopathic asthenozoospermia and teratozoospermia;Yishen Tongluo prescription;syndrome of kidney deficiency and collateral obstruction;efficacy evaluation
摘要:ObjectiveTo carry out a health technology assessment (HTA) of acupuncture for secondary dysmenorrhea (SD) caused by adenomyosis and endometriosis, in order to provide a reference for relevant medical decision-making.MethodsFrom the perspective of the health system, the assessment covers seven areas, including the technical characteristics, safety, effectiveness, economics, ethical fairness, organizational adaptability, and impact on patients and society. The results are reported accordingly.ResultsThe operational specifications of acupuncture are standardized, and the conditions for its use are clearly defined. Acupuncture has a lower overall incidence of adverse events. The main adverse events are localized pain, subcutaneous bleeding, and dizziness, with most symptoms being mild, all of which have corresponding standard treatments. No reports on occupational or environmental safety were found, and the safety operation specifications are available for reference. Compared with conventional Western medicine, acupuncture demonstrates higher effectiveness. Acupuncture may improve the quality of life scores of patients, though no significant difference was observed. The cost of acupuncture is higher than that of conventional Western medicine, but its overall economic value is greater. The informed consent information is relatively comprehensive. Most patients are aware of the potential benefits and risks of acupuncture and voluntarily opt for it. The treatment process fully respects patient privacy and human rights. The clinical application of acupuncture follows the current acupuncture medical service model, with no special requirements for the level of medical institutions. Patient accessibility and affordability are suitable. Patient satisfaction is high. Most patients indicated they would choose acupuncture again for SD. The main barriers to choosing acupuncture are psychological factors (such as fear of acupuncture), cost, and transportation issues. Nearly 70% of patients receiving acupuncture treatment benefit from medical insurance reimbursement, with reimbursement rates generally above 50%, indicating strong social security support.ConclusionThe implementation of HTA for acupuncture in the treatment of SD, using the standards for traditional Chinese medicine (TCM), is feasible. The implementation steps are clear, the data sources for each evaluation domain are adequate, the analysis methods are practical, and the evaluation results are comprehensive. Experts recommend that the findings be used as a reference for relevant medical decision-making.
摘要:ObjectiveTo elucidate the chemical composition of Danshenyin and its blood components in rats after oral administration.MethodsUltra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS/MS) coupled with PeakView 1.2 software was used to systematically characterize and identify the components of Danshenyin aqueous extract and its migratory components in rat blood after oral administration based on the retention time, quasi-molecular ion peaks, secondary fragmentation ions, and literature reports, and a preliminary compounds identification of Salviae Miltiorrhizae Radix et Rhizoma aqueous extract, the co-decoction of Santali Albi Lignum and Amomi Fructus was carried out to attribute the chemical constituents of the aqueous extract of Danshenyin.ResultsA total of 73 compounds, including 21 phenolic acids, 23 diterpenes, 6 flavonoids, 7 organic acids, 3 volatile oils and 13 others, were identified from the aqueous extract of Danshenyin. And 36 prototypes and 15 metabolites were identified in rat plasma, the major metabolic pathways included reduction, hydration, hydroxylation, demethylation, methylation, sulfation and others, these metabolites were mainly derived from tanshinones and salvianolic acids.ConclusionThe main blood components of the aqueous extract of Danshenyin are salvianolic acids and tanshinones, which may be the material basis of the efficacy. This study can provide reference for pharmacological research, quality control, and clinical application of Danshenyin.
关键词:Danshenyin;chemical constituent;serum pharmacochemistry;metabolites;fragmentation pattern;ultra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS/MS)
摘要:ObjectiveStudies have shown that nuciferine has anti-cerebral ischemia effect, but the specific mechanism of action has not been elaborated. Based on the transcriptome results, the pharmacological mechanism of nuciferine against cerebral ischemia was analyzed from multiple dimensions including tissue, cell, pathological process, biological process and signaling pathway.MethodsThirty SD rats were randomly divided into the sham group, model group and nuciferine group(40 mg·kg-1) according to weight. Except for the sham group, the model of middle cerebral artery occlusion(MCAO) was established by thread embolization method after 30 min of administration in the other two groups. Twenty-four hours after surgery, transcriptome sequencing was used to detect the gene expression profiles in the cortex penumbra of rat cerebral tissue, and gene ontology(GO) and kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analysis were performed for differentially expressed genes. The mechanismof nuciferine against cerebral ischemia was analyzed from 5 dimensions of tissue, cell, pathological process, biological process and signaling pathway by the transcriptome-based multi-scale network pharmacology platform(TMNP).ResultsTranscriptome sequencing and gene quantitative analysis showed that 667 genes were significantly reversed by nuciferine. Further enrichment analysis of KEGG and GO suggested that the pathways of nuciferine involved regulating stress response, ion transport, cell proliferation and differentiation, and synaptic function. TMNP research found that at the tissue level, nuciferine could significantly improve the cerebral tissue injury caused by ischemia. At the cellular and pathological levels, nuciferine could play an anti-cerebral ischemia role by improving the state of various nerve cells, mobilizing immune cells, regulating inflammation. And at the level of biological processes and signaling pathways, nuciferine mainly acted on the processes such as vascular remodeling, inflammation-related signaling pathways, and synaptic signaling.ConclusionCombined with the results of transcriptome sequencing, gene quantitative analysis and TMNP, the mechanism of nuciferine against cerebral ischemia may be related to processes such as intervening in stress response and inflammation, affecting vascular remodeling and regulating synaptic function. These results can provide a basis and reference for further study of the pharmacological mechanism of nuciferine against cerebral ischemia.
摘要:ObjectiveTo establish the quality standards for Sennae Folium(Cassia angustifolia) dispensing granules based on standard decoctions.MethodsHigh performance liquid chromatography(HPLC) specific chromatograms were established for 15 batches of Sennae Folium(C. angustifolia) standard decoctions and 10 of Sennae Folium(C. angustifolia) dispensing granules from different manufacturers, and the similarity evaluation, hierarchical cluster analysis(HCA) and principal component analysis(PCA) were performed. Linear calibration with two reference substances(LCTRS) and quantitative analysis of multi-components by single-marker(QAMS) were established for the common peaks in the specific chromatograms to determine the contents of main components in the decoction pieces, standard decoctions and dispensing granules, and to calculate their transfer rates from decoction pieces to standard decoctions and dispensing granules.ResultsThe similarities of specific chromatograms of 15 batches of Sennae Folium(C. angustifolia) standard decoctions and 10 batches of Sennae Folium(C. angustifolia) dispensing granules were all greater than 0.95, and a total of 8 characteristic peaks were calibrated, and five of them were identified, including kaempferol-3,7-O-diglucoside, apigenin-6,8-di-C-glucoside, quercetin-3-O-gentianoside, sennoside B and sennoside A. HCA and PCA results showed that there were certain differences in the composition of different batches of standard decoctions, but no clustering was observed in the production area. As the standard decoctions, the extract rate of 15 batches of samples was 26.54%-45.38%, the contents of kaempferol-3,7-O-diglucoside, apigenin-6,8-di-C-glucoside, quercetin-3-O-gentianoside, sennoside B and sennoside A were 12.16-19.26, 2.57-4.94, 3.27-5.11, 6.75-11.39, 4.69-7.79 mg·g-1, and their transfer rates from decoction pieces to standard decoctions were 45.41%-79.02%, 29.12%-55.07%, 40.52%-67.90%, 24.72%-49.12%, 27.54%-49.34%, respectively. The extract rates of Sennae Folium(C. angustifolia) dispensing granules(C8-C10) were 38.10%-39.50%, the transfer rates of the above five components from decoction pieces to dispensing granules were 72.85%-73.58%, 53.43%-53.94%, 40.19%-40.74%, 24.62%-25.00%, 28.65%-29.11%, respectively, which were generally consistent with the transfer rates from decoction pieces to standard decoctions.ConclusionCompared with the relative retention time method, LCTRS has higher prediction accuracy and is more suitable for chromatographic columns. The established quality control standard of Sennae Folium(C. angustifolia) dispensing granules based on standard decoction is reasonable and reliable, and all indicators of samples from different manufacturers are within the range specified based on the standard decoction, which can provide reference for the quality control and process research of this dispensing granules.
关键词:Sennae Folium;standard decoction;dispensing granules;linear calibration with two reference substances(LCTRS);quantitative analysis of multi-components by single-marker(QAMS);specific chromatograms;quality control;high performance liquid chromatography(HPLC)
摘要:ObjectiveTo study the changing law of appearance color and physicochemical properties of Angelicae Sinensis Radix Carbonisata(ASRC) during the processing by color space method combined with statistical analysis, so as to provide reference for determining the processing endpoint and evaluating the quality of the decoction pieces.MethodsTaking processing time(4, 8, 12, 16 min) and temperature(180, 200, 220, 240 ℃) as factors, ASRC decoction pieces with different processing degrees were prepared in a completely randomized design. Then, the brightness value(L*), red-green value(a*), yellow-blue value(b*), and total chromaticity value (E*ab) of the decoction pieces were determined by spectrophotometer, the color difference value(ΔE) was calculated, and the data of colorimetric values were analyzed by discriminant analysis. At the same time, the pH, charcoal adsorption, and contents of tannins, 5-hydroxymethylfurfural(5-HMF), tryptophan, chlorogenic acid, ferulic acid, senkyunolide I, senkyunolide H and ligustilide of ASRC with different processing degrees were determined by pH meter, ultraviolet and visible spectrophotometry and ultra-high performance liquid chromatography(UPLC). Principal component analysis(PCA) was used to analyze the data of physicochemical indexes, after determining the processing technology of ASRC, the canonical discriminant function was established to distinguish the decoction pieces with different processing degrees, and leave-one-out cross validation was conducted. Finally, Pearson correlation analysis was used to explore the correlation between various physicochemical indexes and chromaticity values.ResultsWith the prolongation of the processing time, L*, a*, b* and E*ab all showed a decreasing trend, and the established discriminant model based on color parameters was able to distinguish ASRC with different processing degrees. The pH showed an increasing trend with the prolongation of processing time, and the charcoal adsorption, and the contents of tannins, 5-HMF, and tryptophan all showed an increasing and then decreasing trend. Among them, the charcoal adsorption, contents of tannin and 5-HMF reached their maximum values successively after processing for 8-12 min. While the contents of chlorogenic acid, ferulic acid, senkyunolide I, senkyunolide H and ligustilide decreased with the increase of processing time, with a decrease of 60%-80% at 8 min of processing. Therefore, the optimal processing time should be determined to be 8-12 min. PCA could clearly distinguish ASRC with different processing degrees, while temperature had no significant effect on the processing degree. The 12 batches of process validation results(10 min, 180-240 ℃) showed that except for 3 batches identified as class Ⅱ light charcoal, all other batches were identified as class Ⅲ standard charcoal, and the chromaticity values of each batch of ASRC were within the reference range of class Ⅱ-Ⅲ sample chromaticity values. The correlation analysis showed that the chromaticity values were negatively correlated with pH and charcoal adsorption, and positively correlated with contents of tryptophan, chlorogenic acid, ferulic acid, senkyunolide I, senkyunolide H, and ligustilide. And both pH and charcoal adsorption were negatively correlated with the contents of the above components, but the charcoal adsorption was positively correlated with the content of 5-HMF.ConclusionThe chromaticity values and the contents of various physicochemical indicators of ASRC undergo significant changes with the prolongation of processing time, and there is a general correlation between chromaticity values and various physicochemical indicators. Based on the changes in color and physicochemical indicators, the optimal processing time for ASRC is determined to be 8-12 min. This study reveals the dynamic changes of the relevant indexes in the processing of ASRC, which can provide a reference for the discrimination of the processing degree and the quantitative study of the processing endpoint.
摘要:ObjectiveTo improve the accuracy of discrete element method in simulating the processing of traditional Chinese medicine(TCM) powder system under low shear conditions.MethodsIn this study, extract powders of Tongsaimai tablets and Qige granules were used as the research objects, the angle of repose(AOR) and effective angle of internal friction of the two materials were determined by AOR test method and shear cell test method. Based on the Hertz-Mindlin with JKR V2 contact model and particle scaling theory, taking the particle-particle restitution coefficient(A), particle-particle static friction coefficient(B), particle-particle rolling friction coefficient(C), particle-steel restitution coefficient(D), particle-steel static friction coefficient(E), particle-steel rolling friction coefficient(F) and Johnson-Kendall-Roberts(JKR) surface energy(G) as test factors, the simulated contact parameters of Tongsaimai tablets extract powder were first calibrated with a single reference value using AOR as the reference value, and then the simulated contact parameters of Tongsaimai tablets extract powder as well as Qige granules extract powder were co-calibrated with AOR and effective angle of internal friction as the joint reference value, respectively. Then, Plackett-Burman design was used to screen the critical contact parameters that have a significant effect on the simulated reference value, and the steepest ascent design was used to determine the optimal range of the critical contact parameters, finally, the regression model between the critical contact parameters and the simulated reference values was established through the design of the response surface test, and the critical contact parameters were calibrated based on the regression model and the desirability function approach.ResultsThe optimal combination of discrete elemental contact parameters A-G for Tongsaimai tablets extract powder under a single reference value was 0.100, 0.718, 0.616, 0.100, 0.400, 0.250 and 0.075 J·m-2, which was validated to have relative errors of 0.10% and -8.64% for the simulated AOR and the simulated effective angle of internal friction, respectively. And the optimal combination of discrete elemental contact parameters A-G for Tongsaimai tablets extract powder at the joint reference values was 0.100, 0.682, 0.598, 0.100, 0.521, 0.294 and 0.075 J·m-2, which was verified to have relative errors of 0.10% and -0.18% for the simulated AOR and the simulated effective angle of internal friction, respectively. The optimal combination of discrete elemental contact parameters A-G for Qige granules extract powder at the joint reference values was 0.150, 0.370, 0.330, 0.150, 0.500, 0.500 and 0.100 J·m-2, which was verified to have relative errors of 2.70% and -1.30% for the simulated AOR and the simulated effective angle of internal friction, respectively. Compared with the single reference value method, the joint calibration method not only increased the number of the critical contact parameters for characterizing particle-device interactions, but also was more accurate and reliable.ConclusionCompared with the results of single reference value calibration, the results obtained by the method of joint calibration of discrete element simulation contact parameters with AOR and effective angle of internal friction as the reference values are more accurate, which can provide more accurate and reliable simulation physical property data for the simulation experiments of TCM extract powder under low shear process conditions.
关键词:discrete element method;angle of repose;effective angle of internal friction;parameter calibration;critical contact parameter;low shear;response surface optimization
摘要:Shengjiangsan is a classic formula for treating warm diseases with wide clinical application and accurate efficacy. There are different opinions on the origin of this formula and lacks key information research on this formula. Therefore, in this study, we conducted systematic research into the historic origin, composition, and other key information of this Shengjiangsan. Results showed that Shengjiangsan has different versions, with "Neixian Fufang", "Jiawei Jianghuangwan", "Peizhensan", and "Taijiwan" being the same formula with different names. Shengjiangsan was first recorded as "Neixian Fufang" in Wanbing Huichun written by GONG Tingxian from the Ming dynasty, inherited and developed by YANG Lishan from Qing dynasty, and has been passed down to modern times. Pills and powder are two main forms of Shengjiangsan, and powder has become more popular nowadays. According to the measurement system of Ming and Qing dynasties, the recommended dosage and usage of Shengjiangsan are as follows. For the pill version of Shengjiangsan, Bombyx Batryticatus of 74.6 g, Curcumae Longae Rhizoma of 9.325 g, Cicadae Periostracum of 9.325 g, and Rhei Radix et Rhizoma of 149.2 g were processed into pills for preparation. Single dosage is Bombyx Batryticatus of 1.15 g, Curcumae Longae Rhizoma of 0.14 g, Cicadae Periostracum of 0.14 g, and Rhei Radix et Rhizoma of 2.3 g, with halved dosage applied for children. For the powder version of Shengjiangsan, the dosage varied in accordance with the severity of the disease. Bombyx Batryticatus of 1.84 g, Curcumae Longae Rhizoma of 0.28 g, Cicadae Periostracum of 0.92 g, and Rhei Radix et Rhizoma of 3.68 g were processed into powder for patients with mild symptoms. Bombyx Batryticatus of 2.48 g, Curcumae Longae Rhizoma of 0.37 g, Cicadae Periostracum of 1.23 g, and Rhei Radix et Rhizoma of 4.91 g were processed into powder for patients with severe symptoms. Bombyx Batryticatus of 3.68 g, Curcumae Longae Rhizoma of 1.84 g, Cicadae Periostracum of 0.55 g, and Rhei Radix et Rhizoma of 7.36 g were processed into powder for patients with critical conditions. In this formula, four herbs were ground to fine powder. For patients with mild symptoms, the whole formula was divided into four dosages, and each dosage weighed 6.71 g. The 200 mL yellow rice wine and 18.65 g honey were added, and the solution was stirred and taken cold till full recovery. For patients with severe symptoms, the whole formula was divided into three dosages, and each weighed 8.95 g. 300 mL yellow rice wine and 27.98 g honey were added, and the solution was stirred and taken cold. For patients with critical conditions, the whole formula was divided into two dosages, and each weighed 13.43 g. 400 mL yellow rice wine and 37.3 g honey were added, and the solution was stirred and taken cold. Shengjiangsan has the effect of ascending lucidity and descending turbidity, dissipating wind, and clearing heat. It is specialized in treating severe heat in exterior, interior, and triple energizers in warm diseases and has a wide modern clinical application. In this study, the historic evolution and key information of Shengjiangsan were reviewed and analyzed, and the key information table of Shengjiangsan was attached, serving as a reference for scholars' research and a theoretical basis for its market transformation.
关键词:classic formula;Shengjiangsan;ancient and modern application;key information;textual research;research progress
摘要:This article systematically analyzes the historical evolution of the name, origin, quality evaluation, harvesting, processing and other aspects of Picrorhizae Rhizoma by referring to the medical books, prescription books, and other documents of the past dynasties, combined with relevant modern research materials, in order to provide a basis for the development and utilization of famous classical formulas containing this medicinal herb. The research results indicate that Picrorhizae Rhizoma was first recorded in New Revised Materia Medica from the Tang dynasty. Throughout history, Huhuanglian has been used as its official name, and there are also aliases such as Gehu Luze, Jiahuanglian and Hulian. The main source of past dynasties is the the rhizomes of Picrorhiza kurrooa and P. scrophulariiflora. In ancient times, Picrorhizae Rhizoma was mainly imported by foreign traders via Guangzhou and other regions, and also produced in China, mainly in Xizang. In ancient times, it was harvested and dried in early August of the lunar calendar, while in modern times, it is mostly harvested from July to September, with the best quality being those with thick and crispy rhizomes without impurities, and bitter taste. Throughout history, Picrorhizae Rhizoma was collected, washed, sliced, and dried before being used as a raw material for medicine, it has a bitter and cold taste, mainly used to treat bone steaming, hot flashes, infantile chancre fever, and dysentery. There is no significant difference in taste and efficacy between ancient and modern times. Based on the research results, it is recommended that the rhizomes of P. scrophulariiflora in the 2020 edition of Chinese Pharmacopoeia, or the rhizomes of P. kurrooa, can be used in famous classical formulas containing this medicinal herb, which can be processed according to the processing requirements marked by the original formula. For those without clear processing requirements, the dried raw products are used as medicine.
关键词:Picrorhizae Rhizoma;famous classical formulas;origin;producing area;quality evaluation;processing method;function and indication;herbal textual research
摘要:ObjectiveWith the development of society and the increasing demand for pregnancy at an advanced age, diminished ovarian reserve (DOR) has attracted wider attention. This disease affects the normal menstruation of women and poses a threat to their mental health. Current research on DOR is still in the initial stage, and advancing research progress in DOR necessitates the construction of effective and clinically relevant animal models. Building on existing literature and integrating the diagnosis criteria of DOR in traditional Chinese medicine (TCM) and Western medicine, this study summarized, analyzed, and evaluated existing models, providing a reference for optimizing DOR animal models.MethodsA retrospective analysis of literature on DOR-related animal models was conducted, and the diagnostic criteria of DOR in TCM and Western medicine were sorted out. By means of evaluation methods of animal models, the clinical relevance of each model to TCM and Western medicine was assessed.ResultsDOR animal models included those based on natural factors, iatrogenic factors, immune factors, metabolic factors, and environmental factors. Among them, the model based on iatrogenic factors, especially models induced by Tripterygium wilfordii preparations, cyclophosphamide, and cisplatin, had a high degree of relevance to both Western medical diseases and TCM syndromes and was confirmed as the optimal modeling method for studying DOR at present.ConclusionThe existing DOR modeling methods mostly have a high degree of clinical relevance to Western medical diseases, while the number of DOR animal models with clear TCM syndromes is very limited. TCM plays an indispensable role in exploring DOR treatment methods. Therefore, the development of DOR animal models related to TCM syndromes should be strengthened, further exploring the potential of TCM and providing strong theoretical support for the application of TCM in DOR treatment.
关键词:diminished ovarian reserve;animal model;integration of disease and syndrome
摘要:Pneumonia is an infectious disease with high morbidity and mortality worldwide, and its damage to the body is not limited to the acute phase. The theory of combination of pathogenic factors emphasizes that the combination of new pathogens and residual pathogens in the body leads to the occurrence of diseases, which generalizes the causes of recurrence during pneumonia recovery. During the recovery stage of pneumonia, pathological changes such as disturbance of immune homeostasis, persistent low-grade inflammation, and microvascular damage continue to affect the body function, impair the health and quality of life of patients, and increase the risk of secondary infection. According to the theory of traditional Chinese medicine (TCM), pneumonia is caused by deficiency, and Qi deficiency and blood stasis is the core pathogenesis in the recovery stage. At this time, the body is not full of healthy qi and still has residual pathogens, and thus it is susceptible to external pathogenic factors that lead to disease recurrence. As an important part of the TCM philosophy of treating disease before its onset, prevention of recurrence after recovery emphasizes the need for aftercare in the recovery stage to prevent disease recurrence. Based on the pathogenesis theory of combination of pathogenic factors and the pathogenesis of Qi deficiency and blood stasis, this paper discusses the effect and connotation of TCM in regulating immune inflammation and microvascular damage in preventing recurrence of pneumonia during the recovery stage, aiming to develop new ideas for effective prevention and treatment of pneumonia at this stage.
关键词:combination of pathogenic factors;immune inflammation and microvascular damage;recovery stage of pneumonia;prevention of recurrence after recovery;theoretical discussion
摘要:Idiopathic pulmonary fibrosis (IPF) can be classified as pulmonary collateral disease,and its pathogenesis is mainly characterized by the loss of Qi meridian nourishment,the loss of Yin meridian nourishment,and the formation of blood stasis in the blood vessels. Qi Yin deficiency is the pathological basis that runs through IPF,and obstruction of meridians and collaterals is a key element in the development of the disease. The dysfunction of "spleen being in charge of the defensive function" is closely related to the formation of the pathological pattern of "lung deficiency and collateral stasis" in IPF. The term "spleen being in charge of the defensive function" originated from the Yellow Emperor's Inner Canon. If the spleen is healthy,the Qi will be filled with vitality. Positive energy is stored inside,evil cannot be dried up. Its concept is quite similar to the immune defense function in modern medicine. If the principle of "spleen being in charge of the defensive function" is lost,the key structure and function of the IPF alveolar epithelial barrier may be abnormal,and it can interact with various innate immune cells to promote inflammation and fibrosis processes. Therefore,this article explains the imbalance of immune homeostasis in IPF alveolar epithelium from two aspects:the barrier function of alveolar epithelial cells(AECs) and their interaction with innate immune cells. And based on the theory of "spleen being in charge of the defensive function",using traditional Chinese medicine for strengthening the spleen and nourishing Qi to treat IPF from the perspective of the spleen. This not only strengthens the scientific connotation of "spleen being in charge of the defensive function" in the pathogenesis of IPF,but also provides new research directions and ideas for its future clinical prevention and treatment.
关键词:spleen being in charge of the defensive function;lung deficiency and collateral stasis;idiopathic pulmonary fibrosis;epithelial homeostasis;alveolar epithelial cells
摘要:Lung cancer is the malignant tumor with the highest incidence and mortality rates globally. Current treatment methods for lung cancer primarily include surgery, chemotherapy, targeted therapy, and immunotherapy. However, the main limitations of these treatments are their side effects, the drug resistance, and the economic burden they impose. As a critical cancer pathway, the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) signaling pathway regulates tumor occurrence and development through multiple mechanisms by influencing various downstream targets. Consequently, the JAK/STAT signaling pathway offers a promising avenue for lung cancer treatment research. Numerous studies have demonstrated that the JAK/STAT signaling pathway plays a key role in the proliferation and growth of lung cancer cells, angiogenesis, epithelial-mesenchymal transition (EMT), metabolic alterations, remodeling of the immune microenvironment, and the development of treatment resistance. Traditional Chinese medicine (TCM) has garnered increasing attention due to its minimal side effects, low economic burden, and its potential to enhance efficacy and reduce toxicity when used in conjunction with Western medicine. In addition to traditional Chinese medicine compounds, a growing number of Chinese medicine monomers have come into the spotlight because of their more targeted effects. Numerous studies investigating the regulation of the JAK/STAT signaling pathway by TCM in the treatment of lung cancer have demonstrated that TCM can inhibit the proliferation and invasion of lung cancer cells, tumor angiogenesis, and EMT, improve the inflammatory and immunosuppressive microenvironments, and enhance treatment sensitivity by intervening in the JAK/STAT signaling pathway, thereby impeding the progression of lung cancer. In recent years, the research on the regulation of this pathway by TCM in the treatment of lung cancer has been updated rapidly. However, the summary of these studies has not been updated in time. This review summarizes and reflects on the recent research findings regarding the regulation of the JAK/STAT signaling pathway by TCM to intervene in lung cancer from three aspects, introducing the JAK/STAT pathway, elaborating the mechanism of this pathway in lung cancer, and exploring the intervention of TCM in the treatment of lung cancer through this pathway, to provide more reference for the treatment of lung cancer in the future.
关键词:Janus kinase (JAK)/signal transducer and activator of transcription (STAT);Chinese medicine monomer;Chinese medicine compound;lung cancer;A549 cells;cell cycle;immunity
摘要:Myocardial ischemia-reperfusion injury (MIRI) is one of the major complications of cardiovascular diseases in recent years, seriously threatening the life and health of patients. In recent years, although significant progress has been made in modern medical research on MIRI, due to its complex pathogenesis, adverse drug reactions, and high postoperative risks, there is an urgent need for safer and more effective therapeutic drugs. The pathological effects of MIRI involve inflammatory responses, oxidative stress, apoptosis, and other mechanisms, while Chinese medicine polysaccharide compounds exhibit pharmacological effects such as anti-inflammation, antioxidation, and anti-apoptosis, which are of great significance for improving MIRI. Numerous studies have found that Chinese medicine polysaccharide compounds mainly exert their effects by activating signal pathways such as nerve regenerative factor-1/epidermal growth factor receptor (NRG-1/ErbB), phosphatidylinositol-3-kinase/protein kinase B (PI3K/Akt), nuclear factor E2-related factor 2/heme oxygenase-1 (Nrf2/HO-1), adenosine 5ʹ-monophosphate-activated protein kinase/silent information regulator 1/peroxisome proliferators-activated receptor γ coactivator lα (AMPK/SIRT1/PGC1α), Ras homologous gene/Rho kinase (Rho/ROCK), glycogen synthase kinase 3 (GSK-3), and P38 mitogen-activated protein kinase (p38-MAPK), thereby playing a role in improving MIRI. This paper, based on an extensive review of relevant literature in China and abroad, systematically summarizes the latest research findings on the use of Chinese medicine polysaccharide compounds in the treatment of MIRI, analyzes the mechanisms by which these compounds improve MIRI, and explores new therapeutic directions based on the joint investigation of signaling pathways and metabolomics. The paper emphasizes the importance of interdisciplinary cooperation in promoting the development of new drugs, aiming to provide valuable references for the basic research and clinical application of Chinese medicine polysaccharide compounds, ultimately benefiting the majority of cardiovascular disease patients.
关键词:Chinese medicine polysaccharide;myocardial ischemia-reperfusion injury;anti-inflammatory;anti-apoptosis;anti-oxidation
摘要:Colorectal cancer is a malignant tumor that originates from the epithelial cells of the colon and rectum. It has the third highest incidence and the second highest mortality rate among malignant tumors worldwide. With the rapid development of the economy and the increasing Westernization of dietary habits in China, its incidence in China has been rising year by year. Over the past decade, despite the introduction of numerous treatment methods for colorectal cancer, the efficacy of existing therapies remains unsatisfactory. In recent years, traditional Chinese medicine (TCM) has become a major focus in the treatment of colorectal cancer due to its advantages of multi-target, multi-pathway mechanisms and low toxicity and side effects. Vascular endothelial growth factor (VEGF) is an important angiogenic factor that promotes blood vessel formation, providing nutrients and oxygen for tumor growth. It also increases vascular permeability, allowing tumor cells to easily pass through the blood vessel wall into other tissues, thereby facilitating metastasis. Several studies have shown that TCM can inhibit tumor angiogenesis and lymphangiogenesis, promote tumor cell apoptosis, and inhibit the proliferation of colorectal cancer cells by acting on the VEGF signaling pathway, thereby delaying tumor growth. In recent years, research in this field has been rapidly updated, but there is a lack of relevant summaries, making subsequent literature searches inconvenient. Therefore, this article focuses on the physiological functions of the VEGF signaling pathway, its role in the occurrence of colorectal cancer, and the intervention of TCM on VEGF, providing a supplement and summary of relevant information to offer a reference for future research in this area.
关键词:vascular endothelial growth factor (VEGF);signaling pathway;colorectal tumor;molecular mechanism;traditional Chinese medicine
摘要:Stroke is the main cause of death and disability among adults in China and is characterized by high incidence, disability, mortality, and recurrence rates. The combination of traditional Chinese and Western medicine has great potential in treating stroke and its sequelae. The classic traditional Chinese medicine prescription Da Chengqitang (DCQT) has a long history and proven efficacy in treating stroke. Clinically, DCQT is often used to treat stroke and its sequelae. However, the number and quality of clinical trials of DCQT in treating stroke need to be improved. Because of the insufficient basic research, the active ingredients and multi-target mechanism of action of DCQT remain unclear. Our research group has previously confirmed that DCQT can effectively reverse neurological damage, reduce iron deposition, and downregulate the levels of pro-inflammatory cytokines in the rat model of hemorrhagic stroke. The treatment mechanism is related to the nuclear factor erythroid 2-related factor 2 (Nrf2)-mediated signaling pathway and p38 mitogen-activated protein kinase (MAPK) signaling-mediated microglia activation. To clarify the pharmacodynamic basis and anti-stroke mechanism of DCQT, this article reviews the research progress in the treatment of stroke with DCQT in terms of clinical trials, pharmacodynamic material basis, safety evaluation, and mechanisms of absorbed components. This article summarizes 45 major phytochemical components of DCQT, 11 of which are currently confirmed absorbed components. Among them, emodin, rhein, chrysophanol, aloe-emodin, synephrine, hesperidin, naringin, magnolol, and honokiol can be used as quality markers (Q-markers) of DCQT. The mechanism of DCQT in treating stroke is complex, involving regulation of inflammatory responses, neuronal damage, oxidative stress, blood-brain barrier, brain-derived neurotrophic factor, and anti-platelet aggregation. This article helps to deeply understand the pharmacodynamic basis and mechanism of DCQT in treating stroke and provides a theoretical basis for the clinical application of DCQT in treating stroke and the development of stroke drugs.
摘要:Diabetes mellitus (DM) is a metabolic disease caused by absolute or relative insulin deficiency and reduced insulin sensitivity in peripheral cells, posing a serious threat to global health. Chronic complications arising in the later stages of DM can lead to the decline or even loss of function in multiple organs, including the eyes, heart, liver, kidneys, nerves, and feet, making them the primary cause of mortality in DM patients. Although modern medicine has made some progress in the treatment of these complications, challenges such as high costs and adverse drug reactions remain. Thus, identifying highly effective drugs with minimal adverse effects has become a top priority. Astragalus membranaceus is a shining gem in the treasure trove of Chinese medicine. Numerous studies have shown that its primary active component, astragaloside Ⅳ, possesses various biological activities, including anti-inflammatory, antioxidant, and antiviral effects, as well as benefits for cardiac and cerebral function, nerve conduction, and myocardial protection. Meanwhile, the phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway plays a crucial role in regulating oxidative stress, inflammatory responses, apoptosis, and autophagy. Extensive research has highlighted the significant role of this pathway in various DM complications, leading to widespread studies on its interaction with astragaloside Ⅳ. This review summarizes research findings on how astragaloside Ⅳ alleviates pancreatic cytotoxicity in DM patients by modulating the PI3K/Akt pathway. Additionally, it highlights its protective effects on basic cardiac function, inhibition of retinal cell damage, improvement of cerebral nerve dysfunction, reduction of chronic kidney and liver damage, and mitigation of neurovascular toxicity in the lower limbs. These insights provide a valuable reference for the clinical application of A. membranaceus and its active monomer, astragaloside Ⅳ, in the treatment of DM and its complications.
关键词:astragaloside Ⅳ;phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) molecular pathway;complications of diabetes;progress in pharmacological research
摘要:Acute lung injury (ALI) is a common and highly lethal clinical syndrome characterized by acute progressive respiratory failure. Currently, the treatment of ALI primarily involves respiratory support therapy and symptomatic pharmacotherapy, yet there is still a lack of specific and effective pharmacological treatments. Qingwen Baidu decoction is a traditional Chinese medicine formula that has the effects of clearing heat, removing toxin, cooling blood, and purging fire. Its pharmacological effects include anti-inflammatory, antipyretic, antibacterial, antiviral, sedative, and so on. The flavonoids, phenols, terpenes, and other components contained in this formula have strong pharmacological activity, which can regulate the inflammatory response caused and oxidative stress in ALI and maintain the integrity of alveolar-capillary barrier (ACB) by anti-apoptosis, anti-pathogen infection, and anti-pulmonary fibrosis, thereby improving the pathological changes of lung tissue. Among them, flavonoids have been reported more, and their mechanism of action is complex and diverse. For example, quercetin, luteolin, and baicalin act on multiple important targets, such as signal transducer and activator of transcription 3 (STAT3), mitogen-activated protein kinase 3 (MAPK3), etc. and participate in the regulation of Toll-like receptor 4 (TLR4)/myeloid differentiation primary response 88 (MyD88)/nuclear factor kappa B (NF-κB), nuclear factor erythroid 2-related factor 2(Nrf2)/Kelch-like ECH-associated protein 1 (Keap1), and silent information regulator 1 (SIRT1)/forkhead box protein O1 (FoxO1) signaling pathways, thereby intervening in pathological events such as inflammation, oxidative stress, cell apoptosis, and fibrosis. This paper aims to review the research progress on Qingwen Baidu decoction and its active ingredients in the prevention and treatment of lung injury in the expectation of providing reference for its subsequent pharmacological mechanism research and theoretical support for its clinical application and drug development in the treatment of ALI.
关键词:Qingwen Baidu decoction;lung injury;acute lung injury;active ingredients;mechanism of action
摘要:Osteoporosis (OP) is a systemic metabolic disease with a strong correlation with age. The prevalence of osteoporosis is rising annually as a consequence of the growing issue of population ageing. The current treatments for OP have numerous shortcomings. In contrast, traditional Chinese medicine has a long history and a rich species diversity. Furthermore, recent years have seen an increase in the number of studies examining the anti-OP properties of traditional Chinese medicine. This may provide a safe and effective alternative strategy for the treatment of OP. The zebrafish, due to its favourable optical transparency and high homology with human genes, has been extensively employed as an animal research model in the investigation of human skeletal-related disease mechanisms and drug screening. This paper presents a review of anti-osteoporosis studies of traditional Chinese medicine using zebrafish as a model for osteoporosis. It also provides a summary of the experimental evaluation methods involved in such studies, an analysis of the current status of traditional Chinese medicine in the treatment of osteoporosis using zebrafish as a model, and a summary of the mechanism of action and the signalling pathways involved in traditional Chinese medicine in the anti-osteoporosis treatment of zebrafish. The current research status of Chinese medicine in the treatment of OP was analysed, as well as the mechanism of action of Chinese medicine against OP and the signalling pathways involved. Furthermore, the advantages and disadvantages of various zebrafish modelling methods of OP were compared with those of traditional animal models. The objective of this study is to provide a reference for the evaluation method of the zebrafish model in the study of bone-related diseases, as well as for the study of the mechanism of action of traditional Chinese medicine against OP and for the reference of the research and development of new drugs.
关键词:osteoporosis;zebrafish;traditional Chinese medicine;signalling pathway;drug screening