Abstract:ObjectiveTo explore the effect of Xiao Xianxiongtang (XXXT) on the transforming growth factor (TGF)-β1-induced invasion, metastasis, and epithelial-mesenchymal transition (EMT) of gastric cancer MGC-803 cells and the underlying mechanism.MethodThe molecular docking between XXXT and nuclear factor of activated T cells (NFAT) was performed by CB-DOCK (http://clab.labshare.cn/cb-dock/). The invasion and metastasis model of MGC-803 cells was established with 10 μg·L-1 TGF-β1. MGC-803 cells were classified into blank group, model group, 0.1 g·L-1 XXXT group, 0.2 g·L-1 XXXT group, and 0.4 g·L-1 XXXT group. For further clarifying the key role of Wnt5a/Ca2+/NFAT signaling pathway in the inhibition of XXXT on gastric cancer, MGC-803 cells were transfected with Wnt5a overexpression plasmid, and then the cells were classified into blank plasmid group, Wnt5a-OE group, blank plasmid + XXXT (0.4 g·L-1) group, and Wnt5a-OE + XXXT (0.4 g·L-1) group. Cell viability was determined by cell counting kit-8 (CCK-8) assay, cell invasion and migration ability by Transwell invasion assay and wound healing assay, expression of EMT-related proteins (E-cadherin, N-cadherin, Vimentin, Snail) and Wnt5a/Ca2+/NFAT signaling pathway-related key proteins [Wnt5a, calcineurin (CaN), NFAT1, and p-NFAT1] by Western blot, and changes in intracellular Ca2+ concentration by immunofluorescence assay.ResultMolecular docking suggested that XXXT acted on Wnt5a/Ca2+/NFAT signaling pathway. XXXT (0.1, 0.2, 0.4 g·L-1) significantly promoted the loss of MGC-803 cell viability (P<0.05,P<0.01). It inhibited cells from invading the transwell lower chamber and slowed down the healing of cell wounds in a dose-dependent manner (P<0.05, P<0.01). Moreover, it promoted the expression of E-cadherin while suppressed the expression of N-cadherin, Vimentin, and Snail (P<0.05, P<0.01). Further experiments showed that XXXT could inhibit the expression of Wnt5a, CaN, NFAT1, and p-NFAT1, and reduce the nuclear expression of NFAT1 and the transcription activity mediated by NFAT1, so as to reduce the cellular Ca2+ concentration (P<0.05, P<0.01). XXXT can reverse the effect of Wnt5a (P<0.05, P<0.01).ConclusionXXXT can attenuate the invasion, metastasis, and EMT of MGC-803 cells via the Wnt5a/Ca2+/NFAT pathway, thereby weakening the tumor-promoting effect of TGF-β1. In summary, XXXT may exert therapeutic effect on gastric cancer by regulating the invasion, metastasis, and EMT of gastric cancer cells.
Keywords:Xiao Xianxiongtang;gastric cancer;invasion and metastasis;epithelial-mesenchymal transition;Wnt5a/Ca2+/ nuclear factor of activated T cells (NFAT) signaling pathway
Abstract:ObjectiveTo explore the molecular mechanism of "transmission between the lung and brain" of influenza based on Janus kinase 1/signal transducer and activator of transcription 1(JAK1/STAT1) signaling pathway and further investigate the intervention effect of Maxing Shigantang (MXSGT).MethodA total of 100 SPF BALB/c mice were randomly divided into a normal group,a model group,an oseltamivir group (21.63 mg·kg-1·d-1),an antiviral granules group(3.9 g·kg-1·d-1), and an MXSGT group(6.05 g·kg-1·d-1), with 20 mice in each group. The pneumonia model was induced in mice except for those in the normal group by intranasal infection of influenza A virus(IAV). Twenty-four hours after modeling,mice were treated with corresponding drugs, while those in the normal group and the model group received the same amount of normal saline by gavage, once a day for 3 and 7 days. The pathological changes in the lung and brain were observed by hematoxylin-eosin(HE)staining. The mRNA expression of IAV nucleoprotein(NP),JAK1, and STAT1 in the lung and brain was detected by real-time quantitative polymerase chain reaction(Real-time PCR), and the protein expression of JAK1 and STAT1 in the lung and brain was detected by Western blot. Immunohistochemical method was used to detect the expression of phosphorylated(p)-STAT1 in the lung and brain tissues, and enzyme-linked immunosorbent assay(ELISA) was used to detect the serum levels of interleukin-1β(IL-1β) and interleukin-10(IL-10).ResultCompared with the normal group, the model group showed obvious pathological changes in the lung tissues and cerebral cortex, increased relative mRNA expression of IAV NP in the lung (P<0.01), elevated mRNA and protein expression of JAK1 and STAT1 in the lung and brain tissues (P<0.05,P<0.01),up-regulated expression level of p-STAT1 in lung tissues and cerebral cortex (P<0.05,P<0.01), and increased serum level of IL-1β (P<0.05). Compared with the model group, the MXSGT group showed alleviated pathological damage to lung tissues and cerebral cortex, decreased relative mRNA expression of IAV NP in lung tissues(P<0.01),reduced mRNA and protein expression levels of JAK1 and STAT1 in lung tissues and brain tissues(P<0.05,P<0.01), and increased serum level of IL-10(P<0.01).ConclusionThe abnormal activation of the JAK1-STAT1 signaling pathway may be one of the molecular mechanisms of "transmission between the lung and brain" of influenza. As an effective compound prescription against the influenza virus,MXSGT can alleviate the pathological damage of brain tissues in mice infected with IAV by regulating the level of cytokines mediated by this pathway.
Keywords:influenza A virus (IAV);"transmission between the lung and brain" of influenza;Maxing Shigantang;Janus kinase 1/signal transducer and activator of transcription 1(JAK1/STAT1) signaling pathway;cytokines
Abstract:ObjectiveTo observe the effects of Sinisan on behaviors and NOD-like receptor protein 3 (NLRP3) inflammasomes of depressed rats induced by chronic unpredictable mild stress (CUMS) and further explore the anti-depressant mechanism of Sinisan.MethodFifty male rats were randomly divided into a normal group, a model group, an NLRP3 inhibitor (MCC950) group (10 mg·kg-1), and low- (2.5 g·kg-1) and high-dose (5 g·kg-1) Sinisan groups, with 10 rats in each group. The depression model was induced by 42 d CUMS in rats except for those in the normal group. Drug intervention was performed on the 22nd day of modeling by gavage in the Sinisan groups and by intraperitoneal injection in the MCC950 group. Except for the MCC950 group, the remaining four groups received 10 mg·kg-1 physiological saline by intraperitoneal injection, while the rats in the model group, the normal group, and the MCC950 group were administered with 3 mL of physiological saline by gavage. Twenty-one days later, the sucrose preference test and open field test were performed. Western blot was used to detect the protein expression of NLRP3, apoptosis-associated speck-like protein containing a CARD (ASC), cysteinyl aspartate-specific protease-1 (Caspase-1), B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), ionized calcium-binding adapter molecule 1 (Iba1), and CD68 in the hippocampus of rats in each group. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of interleukin-1β (IL-1β) and interleukin-18 (IL-18) in the hippocampus of rats. Nissl staining and TUNEL were used to assess the pathological changes and apoptosis level in the hippocampal CA1 region of rats, respectively.ResultThe sucrose preference rate and consumption volume in the sucrose preference test, the total distance, the percentage of central movement distance, and the percentage of residence time in the open field test of rats in the model group were lower than those in the normal group (P<0.01). Compared with the model group, the Sinisan groups and the MCC950 group showed improved depression-like behaviors, apoptosis level in the hippocampal CA1 region, and neuron loss to varying degrees. Sinisan could reduce the levels of IL-1β, IL-18, Bax, Iba1, and CD68 in the hippocampus (P<0.05, P<0.01), increase the level of Bcl-2 (P<0.05, P<0.01), and inhibit the protein expression of NLRP3, ASC, and Caspase-1 related to NLRP3 inflammasomes (P<0.05, P<0.01).ConclusionSinisan can improve the depression-like behaviors and pathological damage of hippocampal neurons in CUMS-induced rats, and the mechanism may be related to the inflammatory response mediated by the NLRP3 inflammasome signaling pathway.
Keywords:Sinisan;chronic unpredictable mild stress (CUMS);neuroinflammation;NOD-like receptor protein 3 (NLRP3) inflammasome;depression;apoptosis
Abstract:ObjectiveTo establish a traditional Chinese medicine (TCM) syndrome model with yin deficiency and internal heat, discuss the rationality of model evaluation, and analyze differentially expressed genes in multiple dimensions to explore the molecular mechanism-signaling pathways as well as key targets of Baihe Dihuangtang (BHDH) in treating depression with Yin deficiency and internal heat.MethodForty male SD rats were randomly divided into a blank control group,a model group,a fluoxetine group (positive drug),a BHDH group, and a Zhibai Dihuangtang group (positive drug for Yin deficiency and internal heat). The depression model with Yin deficiency and internal heat was induced by chronic unpredictable mild stress (CUMS)combined with Chinese herbal drugs with warm and heat nature. The model established was comprehensively evaluated by the detection of the basic condition, behavioral performance, and biochemical indicators of rats in each group. The differentially expressed genes were screened out by mRNA sequencing and underwent Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analyses. The protein-protein interaction (PPI) network was plotted and key genes were analyzed to explore the underlying mechanism of BHDH in treating depression with Yin deficiency and internal heat.ResultThe comparison of basic conditions, behavioral assays, energy metabolism, endocrine hormones, cytokines, and neurotransmitters showed that the model was properly induced. BHDH could significantly improve depression with Yin deficiency and internal heat by regulating the pathways related to the nervous system, endocrine system, and inflammatory and immune system. The key genes of the PPI network were Fos, Epha8, Npy2r, Htr2c, and Nr4a1.ConclusionUnder the guidance of TCM theories of treatment based on syndrome differentiation and etiology and pathogenesis,this study established a depression model with yin deficiency and internal heat in animals and evaluation system in accordance with the symptoms and signs of emotional diseases, and further confirmed the scientificity of the modeling method and the underlying mechanism of BHDH in interfering with depression with Yin deficiency and internal heat based on the results of mRNA sequencing.
Keywords:emotional diseases;depression;model with Yin deficiency and internal heat;Baihe Dihuangtang;mechanism researc
Abstract:Zuojinwan originated from Danxi′s Experiential Therapy (《丹溪心法》) in the Yuan dynasty. It is a representative prescription for the treatment of liver fire invading stomach syndrome, and is also one of the typical prescriptions of the anti-adjuvant method of traditional Chinese medicine (TCM). In this paper, the method of bibliometrics was used to systematically sort out the ancient books of Zuojinwan, and 729 relevant literature data were obtained. After certain retrieval and screening, 57 ancient books of TCM were finally obtained. The statistics and analysis were carried out from the aspects of prescription source, historical evolution, composition, functions, evolution of prescription meaning, prescription dose, and preparation and usage of Zuojinwan. It was found that Zuojinwan was composed of Coptis chinensis rhizoma and Euodia rutaecarpa fructus in a ratio of 6∶1. It was mainly used for the treatment of liver fire invading stomach syndrome. The symptoms included pain in chest and hypochondrium, vomiting and bitter mouth, noisy acid-swallowing, red tongue coating yellow, and pulse string number. Later medical records recorded that Zuojinwan was mostly consistent with the original prescription. It mainly treated various diseases caused by liver fire, including left by liver fire, including left hypochondriac pain, swallowing acid and vomiting acid, tendon hernia and lump, epigastric pain, bitter mouth pulse string, head pain, diarrhea, gonorrhea, cold and hot, abdominal pain, alcohol wet yellowing, silence of oral dysentery and so on. There was little controversy in the analysis of relevant prescriptions. In the past dynasties, pills was mainly used, which was consistent with the original prescription. In modern times, it is mainly water flooding for pills or steamed cakes for pills, warm boiling water to serve 6 g, taking 2-3 g per time, the history is basically the same. In this paper, through the excavation, collation and systematic analysis of the ancient literature of TCM that recorded Zuojinwan, we hope to provide the literature basis for the development, inheritance and utilization of this famous classical formulas.
Keywords:Zuojinwan;Danxi′s Experiential Therapy;literature research;indications and symptoms;Coptidis Rhizoma;Euodiae Fructus;famous classical formulas
Abstract:ObjectiveTo observe the pathological changes of hepatic sinusoidal obstruction syndrome (HSOS) induced by different doses of monocrotaline (MCT) in rats, investigate the dose and duration of modeling, and elucidate the mechanism.MethodA total of 72 male SD rats were randomized into normal group (n=12), and low-, medium-, and high-dose MCT groups (n=20 per group, 80,120,160 mg·kg-1, respecctively). In the model groups, different doses of MCT were intragastrically administered to induce the HSOS in rats. After 48 h and 120 h separately, rats in each group were sacrificed and sampling was performed. The survival rate of rats in each group was calculated, and the body weight, liver weight, and and serum liver function indexes of the rats were examined. The histopathological changes of the liver were observed based on scanning electron microscopy, hematoxylin and eosin (HE) staining, and Sirius red (SR) staining. Glutathione S-transferase (GST) activity, total superoxide dismutase (T-SOD) activity, and malondialdehyde (MDA) content of liver tissue homogenate were measured with microplate method. The expression of liver tissue-related indexes was detected by real-time polymerase chain reaction (PCR), Western blot, and immunohistochemistry.ResultThe activity of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in MCT groups rose with the increase in MCT dose (P<0.05, P<0.01) compared with that in the normal group. With the extension of modeling time, the activity of serum ALT and AST in the low-dose group decreased (P<0.01), while the activity of them in the medium-dose and high-dose groups increased (P<0.01). HE staining showed that hepatocyte necrosis, inflammatory cell infiltration, and erythrocyte accumulation in MCT groups. Electron microscopy demonstrated that fenestrae of liver sinusoidal endothelial cells widened and the sieve plates disappeared. Morever, the injury was worsened with the increase in MCT dose. In addition, the expression of CD44 in MCT groups was significantly reduced compared with that in the normal group (P<0.05, P<0.01). SR staining showed that no positive staining was found in model groups after 48 h, while collagen deposition in portal areas and liver sinusoids could be seen in model groups after 120 h. MCT groups showed increase in MDA content and GST activity and decrease in T-SOD activity compared with the normal group, particularly the medium-dose and high-dose groups (P<0.01), and the changes were dose-dependent after 120 h (P<0.01). The protein expression of CD68 (pro-inflammatory macrophage marker) was raised with the increase in dosage, which was consistent with the results of immunohistochemistry (P<0.01), while CD163 (anti-inflammatory macrophage marker) protein and mRNA expression was significantly decreased with the increase in dosage (P<0.01). Western blot results showed that the expression of phosphorylated nuclear factor-κB/nuclear factor-κB (p-NF-κB/NF-κB) and phosphorylated protein kinase B/protein kinase B (p-Akt/t-Akt) was significantly increased in medium-dose and high-dose MCT groups (P<0.05,P<0.01). The protein expression of α-smooth muscle actin (α-SMA) in liver tissues in MCT groups was significantly increased over time and with the increase in dose, and the mRNA expression of α-SMA, collagen type I α1 (Col1a1), and collagen type Ⅳ α1 (Col4a1) showed the same trend (P<0.05, P<0.01). The results of TUNEL staining showed that apoptotic cells were increased with the rise of MCT dose, while B-cell lymphoma-2(Bcl-2) /Bcl-2 associated X protein (Bax) was remarkably decreased (P<0.01).ConclusionHSOS in rats induced by intragastric administration of different doses of MCT was aggravated with the increase of dosage. In the low-dose (80 mg·kg-1) MCT group, the liver healed spontaneously over time. However, liver damage caused by MCT of 120 mg·kg-1 and 160 mg·kg-1 aggravated over time, and even fibrosis and death occurred. The pathological mechanism of MCT-induced HSOS in rats may be that MCT triggered intense oxidative stress in liver tissue, thus activated pro-inflammatory macrophages to secrete large amounts of inflammatory factors, and further activated the NF-κB/Akt signalling pathway, leading to severe cell damage and death.
Keywords:monocrotaline;hepatic sinusoidal obstruction syndrome;oxidative stress;macrophages;nuclear factor-κB/protein kinase B(NF-κB/Akt) signal
Abstract:ObjectiveTo explore the intervention effect of Erxian decoction on intestinal microflora after ovariectomy in rats by 16S rRNA gene sequencing.MethodThirty-two female healthy SD rats were randomly divided into a Sham operation (Sham) group, a model (OVX) group, an estrogen (E) group, and an Erxian decoction (EXD) group, with 8 rats in each group. The rats in the E group and the EXD group received 1.8×10-4 g·kg-1 estradiol valerate solution and 9 g·kg-1 Erxian decoction, respectively, and those in the Sham group and the OVX group received an equal volume of distilled water once a day for 16 weeks. After 16 weeks, the levels of serum estrogen and blood lipid were detected. The fecal DNA was extracted, followed by 16S rRNA gene sequencing and analysis.ResultCompared with the Sham group, the OVX group showed reduced serum estrogen level (P<0.01) and increased serum levels of total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) (P<0.05). Compared with the OVX group, the E group and the EXD group showed increased serum estrogen level (P<0.01) and reduced TC and LDL-C (P<0.05). Alpha diversity showed that there was no significant change in intestinal microflora diversity after ovariectomy. Beta diversity showed that there were significant differences in the structure of intestinal microflora in the four groups. The intervention of Erxian decoction could improve the changes in intestinal microflora after ovariectomy. LEfSe was used to analyze the differential flora in the four groups. The results showed that the Sham group and the OVX group had 3 differential bacterial phyla and 18 differential bacterial genera, the OVX group and the E group had 1 differential bacterial phylum and 12 differential bacterial genera, and the OVX group and the EXD group had 3 differential bacterial phyla and 5 differential bacterial genera. Estrogen intervention could reverse the change trend of Ruminococcus 1, Anaerovibrio, and Turicibacter in the OVX group. Erxian decoction intervention could reverse the change trend of Bacteroidetes, Firmicutes, Prevotella 9, Ruminococcaceae UCG-014, Ruminococcus 1, and Fusicatenibacter in the OVX group.ConclusionThe structure and function of intestinal microflora in ovariectomized rats changed obviously, and Erxian decoction could ameliorate the change.
Abstract:ObjectiveTo observe the effects of Scutellariae Radix (SR)-Paeoniae Radix Rubra (PRR) combination of different proportions on the expression of the Toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88)/nuclear transcription factor κB (NF-κB) and phosphatidylinositol kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathways in liver tissues of rats with hepatic fibrosis and explore the mechanism against hepatic fibrosis.MethodSixty male SD rats of SPF grade were randomly divided into a normal group, a model group, a positive control (silymarin) group, and SR-PRR 1∶1, SR-PRR 1∶2, and SR-PRR 1∶4 groups, with 10 rats in each group. The hepatic fibrosis model was induced in rats except for those in the normal group by intraperitoneal injection of 40% tetrachloromethane (CCl4)-olive oil solution at 3 mL·kg-1, 5 mL·kg-1 for the first time, for 8 weeks, twice per week. After 4 weeks, rats were treated correspondingly at 10 mL·kg-1 by intragastric administration, and the body weight of rats in each group was weighed for 8 weeks. After administration, histopathological changes in the liver were observed. Serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), hyaluronic acid (HA), laminin (LN), albumin (ALB), alkaline phosphatase (AKP), and superoxide dismutase (SOD) activities, malondialdehyde (MDA), and hydroxyproline (HYP) content in liver tissues were detected. The mRNA expression levels of TLR4, MyD88, NF-κB, PI3K, Akt, and mTOR in the liver of rats were detected by real-time fluorescence-based quantitative polymerase chain reaction (Real-time PCR).ResultCompared with the model group, SR-PRR combination of different proportions could recover the body weight and improve the pathological injury of the liver. As revealed by enzyme linked immunosorbent assay (ELISA) results, compared with the normal group, the model group showed increased ALT, AST, HA, LN, AKP, MDA, and HYP levels to different degrees (P<0.05). Compared with the model group, the groups with drug intervention showed decreased levels of ALT, AST, HA, LN, AKP, MDA, and HYP, potentiated SOD activity, and increased level of ALB (P<0.05). As revealed by Real-time PCR results, compared with the normal group, the model group showed increased mRNA expression of TLR4, MyD88, NF-κB, PI3K, Akt, and mTOR (P<0.05). Compared with the model group, the groups with drug intervention showed reduced mRNA expression of TLR4, MyD88, NF-κB, PI3K, Akt, and mTOR in the liver of rats (P<0.05).ConclusionSR-PRR combination of different proportions can improve the histopathological injury in liver tissues caused by CCl4, with the optimal effect observed in the SR-PRR 1∶4 group. SR-PRR may inhibit the development of liver fibrosis by inhibiting the expression of TLR4/MyD88/NF-κB and PI3K/Akt/mTOR signaling pathways, thereby alleviating chemical-induced liver injury.
Abstract:ObjectiveTo investigate the possible mechanism of Bushen Zhuyun prescription (BSZYP) to reduce the level of ovarian apoptosis in Brown Norway (BN) rats with luteal phase deficiency (LPD).MethodFifty SPF female BN rats were randomly divided into a model group, a dydrogesterone group (0.002 g·kg-1), and a low (4.5 g·kg-1), a medium (9 g·kg-1), and a high-dose (18 g·kg-1) BSZYP groups, with ten rats in each group. The rats were administrated with corresponding drugs by gavage, once a day for three estrus cycle. Western blot was used to detected the protein expression levels of c-Jun NH2 terminal kinase (JNK), extracellular signal-regulated kinase (ERK), phosphorylated-ERK (p-ERK), phosphorylated-JNK (p-JNK), p38 mitogen-activated protein kinase (p38 MAPK), phosphorylated-p38 MAPK (p-p38 MAPK ), B-cell lymphoma (Bcl-2), and Bcl-2 associated X protein (Bax) in ovary. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression levels of ERK, JNK, p38 MAPK, Bax, and Bcl-2 in ovary. Enzyme-linked immunosorbent assay (ELISA) was used to detect the serum progesterone (P) and estradiol (E2) levels. Hematoxylin-eosin (HE) staining was used to observe the ovarian tissue morphology.ResultCompared with the model group, the recovery of estrus cycle of rats in all BSZYP groups had statistical significance after 1-circle administration (P<0.05). The ovarian tissue morphology in the low-dose BSZYP group was improved, and that in the medium and high-dose BSZYP groups was significantly improved with clear follicle, less vesicular follicle and atretic follicle, and more granular layers. The number of luteum, especially the fresh luteum, in the medium and high-dose groups was increased with smooth edge and large volume. The mRNA expression level of Bcl-2 was increased in all-dose BSZYP groups, while the mRNA expression level of Bax was significantly decreased in all-dose BSZYP group (P<0.05, P<0.01). The mRNA expression levels of JNK and p38 MAPK were significantly decreased in the high-dose BSZYP group (P<0.01), and the mRNA expression levels of ERK were increased in the low and medium-dose BSZYP groups (P<0.05). The protein expression level of Bcl-2 was significantly increased in the medium and high-dose BSZYP groups (P<0.01), and the protein expression level of Bax was significantly decreased in the all-dose BSZYP groups (P<0.01). No significant difference was observed in the protein expressions of JNK, ERK, and p38 MAPK in the BSZYP groups. The protein expression levels of p-ERK in the ovarian tissues of rats were significantly inoreased in the medium and high-dose BSZYP group (P<0.01), p-JNK, and p-p38 MAPK in the ovarian tissues of rats were significantly decreased in the medium and high-dose BSZYP group (P<0.01). The level of E2 was increased in all-dose BSZYP groups (P<0.05, P<0.01), and the level of P in the medium-dose BSZYP group was increased (P<0.05).ConclusionBSZYP improved the serum sex hormones, restored the estrous cycle, reduced atretic follicle and vesiculation, and maintained luteal morphology and function of BN rats, so as to improve luteal function and treat luteal phase deficiency. The mechanism of BSZYP may be related to reduce the level of ovarian tissue apoptosis in BN rats by regulating MAPKs signaling pathway.
Keywords:Bushen Zhuyun prescription;mitogen-activated protein kinase (MAPKs);luteal phase deficiency (LPD);apoptosis;ovary
Abstract:ObjectiveTo investigate the therapeutic effect and the possible mechanism of Mankuining decoction on dextran sulfate sodium (DSS)-induced ulcerative colitis (UC) in mice.MethodA total of 90 male SPF C57BL/6 mice were randomly classified into normal group, model group, mesalazine group (0.266 g·kg-1), and high-, and medium-, low-dose (20, 10, 5 g·kg-1) Mankuining decoction groups, with 15 rats in each group. Mice, except the normal group, drank 3% DSS solution for 7 days to induce UC, and administration (ig) started on the day of modeling. The model group and the normal group were given equivalent amount of 0.9% normal saline once a day for 7 days. The general conditions of mice were recorded every day and the disease activity index (DAI) was calculated. On the 8th day, mice were killed by cervical dislocation. All the colons and feces were collected. The length of colon was recorded, and the histopathological changes of colon were observed based on hematoxylin-eosin (HE) staining. The content of inflammatory factors in colon was detected by enzyme-linked immunosorbent assay (ELISA), and the changes of intestinal flora in mouse feces were determined based on 16SrRNA sequencing.ResultCompared with the normal group, the model group had severe colon damage, reduction in colon length (P<0.01), increase in DAI (P<0.01), decrease in interleukin-10 (IL-10) and transforming growth factor-β1 (TGF-β1) in colon(P<0.01), rise of interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-17α (IL-17α), and tumor necrosis factor-α (TNF-α) in colon (P<0.05, P<0.01), and decrease in abundance and diversity of intestinal flora. Compared with the model group, mesalazine and high-, medium-, low-dose Mankuining decoction alleviated the colon injury, recovered the length of colon (P<0.01), decreased DAI (P<0.01), increased IL-10 and TGF-β1 in colon (P<0.01), and decreased IL-1β, IL-6, IL-17α, and TNF-α in colon (P<0.01). Moreover, they raised the abundance and diversity of intestinal flora compared with the model group, as manifested by the increase in the abundance of Firmicutes, Akkermansia, Dubosiella, and Blautia and the decrease in the abundance of Bacteroidetes, Muribaculaceae, Clostridia_UCG-014, and Alistipes.ConclusionMankuining decoction has definite effect in treating UC mice, and the effect is positively correlated with the concentration. In addition, different concentration has different influence on the structure of flora. The mechanism is the likelihood that it alleviates the disorder of intestinal flora to restore intestinal immune balance and further promote the recovery of colonic mucosa.
Abstract:ObjectiveTo study the effect of icariin on the proliferative capacity of hepatocellular carcinoma cell line CLC5 and the underlying mechanism.MethodThe targets of icariin were screened out by network pharmacology, and the target network and protein-protein interaction (PPI) network were constructed to predict the possible targets and pathways of icariin. CCK-8 assay was employed to explore the effects of different concentrations (0, 6.25, 12.5, 25, 50 μmol·L-1) of icariin on the viability of CLC5 cells. Further, CLC5 cells were treated with 0, 25, 50 μmol·L-1 icariin, and the effect of icariin on CLC5 cell proliferation was examined by Edu-488 assay and clone formation assay (CFA). Western blot was employed to measure the expression levels of proteins in the protein kinase B (Akt)/glycogen synthase kinase 3β (GSK3β)/cell cycle-dependent kinase (CDK) pathway in the CLC5 cells exposed to different concentrations of icariin.ResultNetwork pharmacological analysis revealed that icariin may inhibit the hepatocellular carcinoma via cell cycle arrest and inhibition of tumor cell proliferation. Compared with the blank group, icariin decreased the viability of CLC5 cells in a time- and concentration-dependent manner (P<0.01) and reduced the positive rate of Edu-488 and the colonies in CFA (P<0.05, P<0.01). Moreover, icariin down-regulated the protein levels of p-Akt, p-GSK3β, CDK4, and CyclinD1 (P<0.05, P<0.01).ConclusionIcariin may block cell cycle to suppress the proliferation of CLC5 cells via inhibiting the Akt/GSK3β/CDK pathway.
Abstract:ObjectiveTo investigate the intervention effect of total glucosides of paeony (TGP) on the renal injury of MRL/lpr mice based on the Toll-like receptor 9 (TLR9)/myeloid differentiation factor 88 (MyD88)/nuclear transcription factor-κB (NF-κB) signaling pathway and explore the immunological mechanism of TGP in preventing and treating systemic lupus erythematosus (SLE).MethodMRL/lpr female mice of SPF grade were randomly divided into a model group, a dexamethasone group (0.15 g·kg-1), and high- (0.078 g·kg-1) and low-dose (0.039 g·kg-1) TGP groups, and female C57BL/6J mice were assigned to a blank group, with 7 mice in each group. Mice in each group were treated with corresponding drugs or normal saline by gavage at the same time every day. After 4 weeks, samples were collected. The kidney and spleen were weighed, and the organ index was calculated. Serum creatinine (SCr) and blood urea nitrogen (BUN) levels in each group were detected by biochemical assay. Hematoxylin-eosin (HE) staining was used to observe the histopathological changes in the kidney. The degree of renal fibrosis was evaluated by Masson staining. The serum levels of interleukin (IL)-2, interferon (IFN)-α, IL-4, and anti-nuclear antibody (ANA) were detected by enzyme-linked immunosorbent assay (ELISA). The mRNA expression of TLR9, MyD88, and NF-κB p65 in renal tissues was detected by real-time quantitative polymerase chain reaction (Real-time PCR). The protein expression of TLR9 and NF-κB p65 in renal tissues was detected by immunofluorescence. The protein expression of TLR9, MyD88, and NF-κB p65 in renal and spleen tissues was tested by Western blot.ResultCompared with the blank group, the model group showed increased SCr, BUN, spleen index, and kidney index (P<0.05), deteriorated pathological injury and fibrosis in renal tissues, elevated serum levels of IFN-α, IL-4, and ANA, decreased level of IL-2 (P<0.05), and up-regulated TLR9, MyD88, and NF-κB p65 mRNA and protein levels in the kidney and spleen (P<0.05). Compared with the model group, the TGP groups displayed reduced SCr, BUN, spleen index, and kidney index (P<0.05), relieved pathological damage and fibrosis in renal tissues, decreased serum levels of IFN-α, IL-4, and ANA (P<0.05), increased level of IL-2, and declining mRNA and protein expression levels of TLR9, MyD88, and NF-κB p65 in the kidney and spleen (P<0.05).ConclusionTGP may inhibit the expression of downstream inflammatory factors to regulate immunity and resist SLE-induced renal injury by regulating the TLR9/MyD88/NF-κB signaling pathway.
Keywords:total glucosides of paeony;systemic lupus erythematosus(SLE);Toll-like receptor 9 (TLR9)/myeloid differentiation factor 88 (MyD88)/nuclear transcription factor-κB (NF-κB) signaling pathway;SLE model mice
Abstract:ObjectiveTo investigate the effects of Gandou decoction (GDD) on the mitophagy of hippocampal neurons in toxic milk (TX) mouse model of Wilson disease and explore the protective mechanism of GDD against neuron injury through the PTEN induced kinase 1 (Pink1) /E3 ubiquitin ligase (Parkin) pathway.MethodSixty mice were randomly divided into a blank group, a model group, a penicillamine group (0.09 g·kg-1), and low- (5.5 g·kg-1), medium- (11 g·kg-1), and high-dose (22 g·kg-1) GDD groups, and treated correspondingly by gavage for 8 weeks. Morris water maze, traction test, and pole test were used for the evaluation of animal behaviors. Hematoxylin-eosin (HE) staining and transmission electron microscopy were used to observe cell apoptosis, ultrastructure, autophagy, and mitochondrial structure. The levels of superoxide dismutase (SOD), reactive oxygen species (ROS), and malondialdehyde (MDA) were detected by enzyme-linked immunosorbent assay (ELISA). Real-time fluorescence-based quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of Pink1, Parkin, autophagy-associated protein Beclin-1, microtubule-associated protein 1 light chain 3Ⅱ (LC3Ⅱ), and p62. Western blot was conducted to detect the protein expression of Pink1, Parkin, Beclin-1, LC3Ⅱ/Ⅰ, and p62.ResultCompared with the blank group, the model group showed prolonged escape latency, decreased times of platform crossing, lower score in the traction test, and longer pole climbing time (P<0.01). Compared with the model group, the medium- and high-dose GDD groups and the penicillamine group showed shortened escape latencies, increased times of platform crossing, higher scores in the traction test, and shortened pole climbing time (P<0.01). Compared with the blank group, the model group displayed severely damaged neurons and increased autophagosomes. Compared with the model group, the medium- and high-dose GDD groups and the penicillamine group showed improved neuron damage and reduced autophagosomes. The levels of ROS and MDA were higher and SOD was lower in the model group than those in the blank group (P<0.01), while the levels of the above indicators were reversed by GDD intervention as compared with the model group (P<0.01). Compared with the blank group, the model group exhibited up-regulated mRNA and protein expression of Pink1, Parkin, LC3Ⅱ, and Beclin-1 and down-regulated p62 (P<0.05). Compared with the model group, the medium- and high-dose GDD groups showed reduced mRNA and protein expression of Pink1, Parkin, LC3Ⅱ, and Beclin-1 and increased p62 (P<0.05, P<0.01).ConclusionGDD can significantly inhibit the excessive mitophagy in neurons of TX mice and protect neurons from damage. The mechanism may be related to the regulation of the Pink1/Parkin pathway.
Abstract:ObjectiveTo identify the protective effect and possible mechanism of Gandou Fumu decoction (GDFMD) on liver fibrosis in mice with Wilson's disease.MethodA total of 50 homozygous TXJ mice were randomly divided into five groups, with 10 mice in each group. Ten wild-type mice were selected as a normal group. The GDFMD high, medium, and low-dose groups were given 13.92, 6.96, 3.48 g·kg-1 of GDFMD, respectively. The penicillamine group were given 0.1 g·kg-1 of penicillamine. The model group and the normal group were given the same volume of 0.9% sodium chloride solution once a day for 4 consecutive weeks. The enzyme-linked immunosorbent assay (ELISA) method was performed to detect serum superoxide dismutase (SOD) activity and malondialdehyde (MDA) content. Corresponding kits were used to detect the mitochondrial adenine triphosphate (ATP) content and Na+-K+-ATPase activity in liver tissues. Hematoxylin-eosin (HE) and Masson staining were used to observe the pathological morphology of liver tissue, and transmission electron microscope was used to observe ultrastructural changes of liver tissues in mice. Western blot was used to detect the c-Jun N-terminal kinase, the phosphorylated protein, and the expressions of Caspase-3, B cell lymphoma-2 (Bcl-2), and Bcl-2 associated X protein (Bax) in c-Jun N-terminal kinase (JNK) signaling pathway.ResultCompared with the normal group, MDA content increased and SOD activity decreased in the model group (P<0.05). Compared with the model group, SOD activities in the GDFMD high-, medium-, and low-dose groups and the penicillamine group significantly increased (P<0.01), and MDA content significantly decreased (P<0.05, P<0.01). Compared with the normal group, ATP content and Na+-K+-ATPase activity significantly decrease in the model group (P<0.05). Compared with the model group, ATP content and Na+-K+-ATPase activity in the GDFMD medium and high-dose groups and the penicillamine group significantly increased (P<0.05, P<0.01). The results of the pathological morphology of liver tissue showed that a large number of liver cells degeneration and necrosis, inflammatory cell infiltration, unclear liver lobule structure, and collagen fiber deposition were observed in the model group. Transmission electron microscopy showed that the number of mitochondria in liver tissues significantly reduced, the mitochondria were locally damaged, and the cristae of mitochondria were broken even disappear in the model group. The pathological morphology of liver tissue and mitochondrial structure recovered to varying degrees after medicinal intervention. The results of Western blot suggested that, compared with the normal group, the expression levels of phosphorylation-JNK (p-JNK), p-JNK/JNK, Caspase-3, and Bax in the liver tissues were up-regulated, while the expression of Bcl-2 was down-regulated in the model group (P<0.05). The expression levels of p-JNK, p-JNK/JNK, Caspase-3 and Bax were down-regulated and the expression of Bcl-2 was up-regulated in the GDFMD high and medium-dose groups and the penicillamine group (P<0.01).ConclusionGDFMD can alleviate oxidative stress damage and recover mitochondrial function of TXJ mice with liver fibrosis. The mechanism of GDFMD may be related to regulating the JNK signaling pathway and downstream factors and inhibiting cell apoptosis.
Abstract:ObjectiveTo observe the clinical efficacy of Gandou Fumu granules (GDFM) in the treatment of Wilson disease (WD) with liver-kidney deficiency and phlegm-blood stasis.MethodNinety WD patients in The First Affiliated Hospital of Anhui University of Chinese Medicine were randomly divided into a control group (45 cases) and a treatment group (45 cases). All patients were treated with sodium 2,3-dimercaptopropane-1-sulfonate (DMPS), while those in the treatment group received additional GDFM. All patients were treated for four courses (32 days). The traditional Chinese medicine (TCM) syndrome scores,clinical effective rate,24 h urinary copper,ceruloplasmin (CER),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6 (IL-6),superoxide dismutase (SOD),glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) levels of the two groups before and after treatment were observed.ResultAfter treatment, the TCM syndrome scores of the two groups decreased (P<0.01),and the score of TCM syndrome in the treatment group was lower than that of the control group (P<0.01). The total effective rate of the treatment group was 82.22% (37/45), higher than 57.78% (26/45) of the control group (χ2=6.402,P<0.05). There was no significant difference in CER before and after treatment in both groups. The post-treatment 24 hour urinary copper increased (P<0.01), which was higher in the treatment group than that in the control group (P<0.05). The TNF-α,IL-1β, and IL-6 levels were significantly reduced in both groups after treatment(P<0.01),and the above indicators in the treatment group were significantly lower than those in the control group (P<0.01). After treatment,the SOD level increased and the MDA level decreased in the control group (P<0.01), while no significant difference in GSH-Px level was observed. The SOD and GSH-Px levels increased and the MDA level decreased in the treatment group (P<0.01). After treatment, SOD and GSH-Px levels of the treatment group were higher than those in the control group, while the MDA level was lower than that in the control group(P<0.05,P<0.01).ConclusionGDFM can improve the TCM syndrome score and clinical efficacy,enhance the copper removing effect,and inhibit the inflammatory response and antioxidative stress in the treatment of WD with liver and kidney deficiency and phlegm-blood stasis.
Keywords:Gandou Fumu granules;hepatolenticular degeneration;Wilson disease;liver and kidney deficiency;phlegm-blood stasis
Abstract:Hepatolenticular degeneration(HLD),also known as Wilson disease (WD), is a genetic disorder characterized by copper metabolism disorder caused by ATP7B gene mutation. Specifically, due to the ceruloplasmin synthesis disorder induced by gene mutation,copper cannot be excreted through bile,which results in pathological deposition of copper in various organs and damage to organs such as the brain and the liver. The incidence of WD in Chinese is significantly higher than that in the world. Copper chelating agents, such as D-penicillamine and dimercaptosuccinic acid, are used as the main therapeutic agents in western medicine. However, many clinical adverse events limit the application of these drugs. Traditional Chinese medicine (TCM) has its characteristics in the treatment of WD. As confirmed by long-term research on TCM clinical diagnosis and treatment,MD has become TCM dominant disease. In spite of many views about the etiology and pathogenesis of WD,a consensus has not been reached so far. Based on the theory of latent pathogen in TCM and the pathological mechanism of excessive deposition of copper ions in the body,this study proposed that latent toxin is the key etiology of WD,and further elaborated that the latent toxin of WD was inherited from parents and occurred in children and adolescents,which was hidden in the liver and the kidney and damaged the brain. The latent toxin, Yang in nature and dispersing in property, is prone to transform into dampness-heat to block Qi movement and produce phlegm leading to stasis. Furthermore, this study determined latent toxin blocking collaterals as the basic pathogenesis of WD and revealed the complex clinical manifestations of latent toxin blocking collaterals such as liver collaterals,brain collaterals,kidney collaterals,spleen collaterals,stomach collaterals,lung collaterals,heart collaterals, and uterus collaterals. Treatment should follow the basic therapeutic principles of resolving pathogens,removing toxins, and dredging collaterals. This study is expected to provide a theoretical basis for syndrome differentiation and treatment of WD in TCM.
Abstract:ObjectiveTo evaluate the clinical curative effect of modified Shaofu Zhuyutang on pelvic pain of endometriosis (EMT) with syndrome of cold congeal and blood stasis and the influence on neural angiogenesis.MethodA total of 110 cases were divided into a control group (54 cases) and an observation group (56 cases) by the random number table method. Patients in the control group took Aifu Nuangong pills with 6 g/time and 2 times/day. Patients in the observation group took modified Shaofu Zhuyutang with 1 dose/day. Course of treatment continued for 3 menstrual cycles. Dysmenorrhea, other symptoms and signs of pelvic pain, and the EMT health profile-5 (EHP-5) for patients with syndrome of cold congeal and blood stasis and EMT were scored before and after treatment. The levels of peripheral blood nerve growth factor (NGF), substance P (SP), calcitonin gene-related peptide (CGRP), brain-derived neurotrophic factor (BDNF), soluble Fms-like tyrosine kinase(sFlt-1), vascular endothelial growth factor (VEGF), matrix metalloproteinase-9 (MMP-9), and insulin growth factor-1 (IGF-1) were detected before and after treatment. The levels of prostaglandin E2 (PGE2), cyclooxygenase-2 (COX-2), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and other inflammatory factors were detected before and after treatment. The pain medication usage and recurrence were recorded.ResultThe scores of dysmenorrhea, symptoms of pelvic pain symptoms (non-menstrual pelvic pain, dyspareunia, anal falling pain, and defecation pain, etc.), and signs of pelvic pain (pelvic tenderness and sacral ligament tender nodules) in the observation group were lower than those in the control group (P<0.01). The disappearance rates of dysmenorrhea, pelvic tenderness, and sacral ligament tender nodules were 67.35% (33/49), 73.33% (33/45), and 77.27% (34/44) in the observation group, which were all higher than 45.83% (22/48), 48.84% (21/43), and 52.27% (23/44) in the control group. The difference was statistically significant (χ2=4.571, χ2=5.565, χ2=6.026, P<0.05). The scores of EHP-5 and syndrome of cold congeal and blood stasis in the observation group were lower than those in the control group (P<0.01). The levels of VEGF, MMP-9, IGF-1, NGF, SP, CGRP, and BDNF in the observation group were lower than those in the control group (P<0.01), while the level of sFlt-1 was higher than that in the control group (P<0.01). The levels of PGE2, COX-2, TNF-α, and IL-6 in the observation group were lower than those in the control group (P<0.01). The total effective rate of the observation group was 92.45% (49/53), which was higher than 76.00% (38/50) of the control group (χ2=5.307, P<0.05). After 6 months of follow-up, the recurrence rate in the observation group was 30.61% (15/49), which was lower than 52.63% (20/38) in the control group (χ2=4.315, P<0.05). The average of taking ibuprofen sustained-release capsules in the control group was higher than that in the observation group per menstrual period (P<0.01).ConclusionModified Shaofu Zhuyutang treated pelvic pain of EMT with syndrome of cold congeal and blood stasis by regulating the mechanism of neural angiogenesis, reducing pain, and promoting the disappearance of related pains, thus improving the quality of life. Shaofu Zhuyutang has a better clinical effect than Aifu Nuangong pills and has a low recurrence rate.
Keywords:endometriosis;pelvic pain;syndrome of cold congeal and blood stasis;modified Shaofu Zhuyutang;neural mechanism;angiogenesis
Abstract:ObjectiveTo investigate the influencing factors of intestinal metaplasia or atypical hyperplasia in chronic atrophic gastritis (CAG) patients and establish a prediction model.MethodThe clinical records and laboratory examination data of 335 CAG patients treated in the department of gastroenterology of the Second Affiliated Hospital of the Anhui University of Chinese Medicine from June 2016 to June 2021 were collected. Single and multiple Logistic regression analyses were used to explore the influencing factors of intestinal metaplasia or atypical hyperplasia in CAG patients by SPSS 26.0. A prediction model was constructed based on the data of the related influencing factors. In addition, 115 CAG patients diagnosed in the First Affiliated Hospital of Anhui Medical University from June 2019 to June 2021 were selected as external validation samples to verify and evaluate the prediction efficiency of the constructed prediction model.ResultMultiple Logistic regression analysis showed that pepsinogen Ⅰ[odds ratio(OR) 0.994,95% confidence interval(CI) (0.990,0.999),P<0.05],the number of focus[OR 6.765,95% CI(3.831,11.945),P<0.01], and Helicobacter pylori (Hp) infection[OR 0.546,95% CI(0.335,0.888),P<0.05] were independent risk factors for intestinal metaplasia or atypical hyperplasia in CAG patients(P<0.05). The formula of the prediction model is as follows:P=-1.558+0.606×Hp infection-0.006×pepsinogen Ⅰ+1.912×the number of focus. The receiver operating characteristic (ROC) curve showed the specific parameters as below: the area under the ROC curve of 0.76,the Youden index of 0.443,the best cut-off value of 0.52,sensitivity of 0.533,and specificity of 0.910. The prediction model was applied to the data of patients in the validation group for validation,and the predictive efficiency of the model was tested by decision curve analysis (DCA). The results showed that the model had a good fit and high predictive value.ConclusionPepsinogen Ⅰ,the number of focus, and Hp infection are independent risk factors for intestinal metaplasia or atypical hyperplasia in CAG patients. The prediction model constructed based on these factors has a good fit and high predictive value,which can provide references for the classification of CAG patients and the formulation of individual treatment protocols.
Keywords:chronic atrophic gastritis;intestinal metaplasia;atypical hyperplasia;influencing factors;prediction model
Abstract:ObjectiveMetabolomics was used to identify biomarkers of chronic alcoholism, and to evaluate the neuroprotective effect of geniposide, providing reference for the diagnosis and treatment of chronic alcoholism.MethodThe rat model of chronic alcoholism was established by intragastric administration of 50% ethanol with 8 mL·kg-1 for 14 days, and then increased to 12 mL·kg-1 for 21 days. Meanwhile, the intervention was performed by continuous gavage of geniposide (15 mg·kg-1) for 35 days. At the end of the experiment, the biochemical indexes and histopathological morphology of liver and brain tissues of rats were detected. Ultra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS) was used for urine metabonomics. The chromatographic conditions was as follows:ACQUITY UPLC™ HSS T3 column (2.1 mm×100 mm, 1.8 μm), mobile phase of 0.1% formic acid acetonitrile solution (A)-0.1% formic acid aqueous solution (B) for gradient elution (0-2.5 min, 1%-11%A; 2.5-4.5 min, 11%-21%A; 4.5-7.0 min, 21%-40%A; 7.0-8.5 min, 40%-99%A; 8.5-10.5 min, 99%A; 10.5-10.6 min, 99%-1%A; 10.6-13.0 min, 1%A), the flow rate of 0.4 mL·min-1. The conditions of mass spectrometry were electrospray ionization (ESI), positive and negative ion modes, scanning range of m/z 50-1 200. Progenesis QI 2.0 and MassLynx 4.1 were used for data analysis, and biomarkers were identified by matching element composition and secondary fragments with Human Metabolome Database (HMDB).ResultThe pathological results showed that on the 35th day of model replication, compared with the model group, the cortical neurons in the geniposide group showed a significantly improved state of disorder, nuclear pyknosis, hyperchromatism and cell membrane boundary blurred necrosis. The biochemical results showed that geniposide could significantly increase the activities of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD), decrease the activity of acetylcholinesterase (AChE), decrease the levels of β-endorphin (β-EP) and malondialdehyde (MDA). A total of 48 biomarkers of chronic alcoholism were identified by metabonomics, involving seven metabolic pathways of tryptophan metabolism, phenylalanine metabolism, pentose and glucuronate interconversions, pyrimidine metabolism, ascorbate and aldarate metabolism, steroid hormone biosynthesis and purine metabolism. The main pathway is 5-hydroxytryptamine pathway of tryptophan metabolism.ConclusionBiomarkers related to nerve injury in chronic alcoholism are mainly derived from the 5-hydroxytryptamine metabolic pathway. Geniposide can regulate this pathway so as to improve oxidative stress in the brain and play a neuroprotective role.
Keywords:geniposide;chronic alcoholism;metabolomics;ultra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS);neuroprotection;tryptophan metabolism;oxidative stress
Abstract:ObjectiveTo analyze the differential components in water extract of Chuanxiong Rhizoma before and after processing with wine, and to explore the molecular mechanism of Chuanxiong Rhizoma processed with wine in enhancing anti-cerebral ischemia injury.MethodUltra high performance liquid chromatography tandem quadrupole orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS) was used to qualitatively analyze the main chemical components in water extract of Chuanxiong Rhizoma based on the spectral information of compound, comparison of reference substance and references. The chemical pattern recognition method was used to screen the differential components of Chuanxiong Rhizoma before and after processing. Based on these differential components, the potential targets of differential components were predicted by online databases, and the related targets of cerebral ischemia were searched. Cytoscape 3.6.0 was used to establish the network diagram of differential components-action targets-diseases of Chuanxiong Rhizoma processed with wine. The protein-protein interaction (PPI) network of intersection targets was constructed by STRING 11.5. The potential targets of differential components against cerebral ischemia were analyzed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis through DAVID 6.8. At the same time, the chemical compounds with high relative content and increased peak area after wine processing were docked with their corresponding targets to verify the mechanism of enhanced effect after wine processing.ResultA total of 71 chemical components were identified from Chuanxiong Rhizoma, 34 differential components and 603 potential targets were screened out. At the same time, a total of 769 disease targets and 60 intersection targets were obtained. Seven key targets were identified through PPI network analysis, including JUN, signal transducer and activator of transcription 3 (STAT3), mitogen-activated protein kinase 3 (MAPK3), interleukin-1β (IL-1β), vascular endothelial growth factor A (VEGFA), Caspase-3 (CASP3) and mtrix metalloproteinase 9 (MMP9). Tumor necrosis factor (TNF) signaling pathway was the main differential signaling pathway. The results of molecular docking showed that differential components (senkyunolide K, senkyunolide F, 3-n-butylphthalide, Z,Z′-6,8′,7,3′-diligustilide, ferulic acid and Z-ligustilide) and corresponding targets had good binding activities.ConclusionThe synergistic mechanism of Chuanxiong Rhizoma processed with wine may be related to the enhanced inhibitory effect of inflammatory reaction.
Keywords:Chuanxiong Rhizoma;wine-processed;cerebral ischemia;ultra high performance liquid chromatography tandem quadrupole orbitrap high resolution mass spectrometry (UHPLC-Q-Orbitrap HRMS);network pharmacology;molecular docking
Abstract:ObjectiveTo predict the possible quality markers (Q-markers) of Arisaema Cum Bile in the prevention and treatment of stroke based on ultra performance liquid chromatography-quadrupole-time-of-flight tandem mass spetrometry (UPLC-QTOF-MS/MS) and Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine (TCMIP) v2.0.MethodUPLC-QTOF-MS/MS was employed with the mobile phase of 0.1% formic acid aqueous solution (A)-0.1% formic acid acetonitrile solution (B) for gradient elution (0-3 min, 0.2%-5%B; 3-5 min, 5%-8%B; 5-8 min, 8%-10%B; 8-14 min, 10%-25%B; 14-18 min, 25%-50%B; 18-20 min, 50%-70%B; 20-21 min, 70%-98%B; 21-23 min, 98%B; 23-24 min, 98%-0.2%B; 24-26 min, 0.2%B), the flow rate of 0.5 mL·min-1 and electrospray ionization (ESI). High quality MS/MS data were scanned in positive and negative ion modes with scanning range of m/z 50-1 500. A local database of the chemical constituents in Arisaema Cum Bile was established by UNIFI 1.8. Then the chemical constituents in Arisaema Cum Bile were characterized by matching with the local database and comparing with the reference substances and literature information. TCMIP v2.0 was used to obtain the targets corresponding to the identified components of Arisaema Cum Bile and stroke, and the "disease-formula" correlation analysis was carried out to screen the core targets by topological eigenvalues. DAVID 6.8 was used for enrichment analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway of core targets. According to the "five principles" of Q-markers and combined with literature reports, the Q-markers of Arisaema Cum Bile in the prevention and treatment of stroke were predicted, and the core components acting on these target genes were obtained. Cytoscape 3.8.0 was employed to draw the network diagram of "medicinal materials-active ingredients-target genes-pathways". Finally, AutoDock Vina 1.2.2 was used to calculate and verify the molecular docking between the candidate components and the key targets.ResultA total of 76 chemical components was identified in positive and negative ion modes, 85 core targets were collected for Arisaema Cum Bile in the prevention and treatment of stroke. A total of 31 stroke-related pathways, 23 target genes and 9 main active components of Arisaema Cum Bile acting on these genes were screened, and then we determined 4 possible Q-markers for Arisaema Cum Bile in the prevention and treatment of stroke according to the "five principles".ConclusionThe possible Q-markers of Arisaema Cum Bile for stroke are gallic acid, apigenin-6,8-di-C-glucoside, apigenin and cholic acid, and the target of these four components may be estrogen receptor alpha (ESR1).
Keywords:Arisaema cum Bile;ultra performance liquid chromatography-quadrupole-time-of-flight tandem mass spetrometry (UPLC-QTOF-MS/MS);estrogen receptor alpha (ESR1);molecular docking;stroke;quality markers (Q-markers);network pharmacology
Abstract:ObjectiveTo conduct phylogenetic analysis of internal transcribed spacer 2 (ITS2) and chloroplast gene segments including psbA-trnH, rbcL, and matK of Sophora japonica cv. jinhuai resource samples from different geographical sources, and to explore the genetic diversity of S. japonica cv. jinhuai.MethodPolymerase chain reaction (PCR) method was used to amplify the nucleic acid sequences of ITS2, psbA-trnH, rbcL, and matK of S. japonica cv. jinhuai. Neighbor joining (NJ) method was used to construct phylogenetic trees, and Kimura 2-Parameter (K2P) model was used to calculate the genetic distance of different samples. MEGA and BIOEDIT softwares were applied for mutiple alignment and analysis of ITS2, psbA-trnH, rbcL, and matK sequences of S. japonica cv. jinhuai.ResultThe lengths of ITS2 sequence were 278-279 bp. The lengths of psbA-trnH were 289 bp. The lengths of rbcL sequence were 673 bp. The lengths of matK sequences were 786-792 bp. There were 3 mutation points in ITS2 and psbA-trnH, no mutation point in rbcL, and 13 mutation points in matK. The samples of S. japonica cv. jinhuai were clustered into two groups based on the phylogenetic tree constructed by ITS2 sequences. The sample of seedling tree in Baibao was clustered into one group, while the other 25 samples were clustered into another group. For the psbA-trnH sequence, the success rate of PCR amplification of 28 samples of S. japonica cv. jinhuai was 100%. The 28 samples of S. japonica cv. jinhuai were clustered into three groups based on the clustering results of psbA-trnH sequence. The sample of seedling tree in Shaoshui was clustered into one group. The five samples of grafting tree and seedling tree in Miaotou, grafting trees in Jiantang, Wenqiao, and Daxu, and seeding tree in Xianshui were clustered into one group. The other 21 samples were clustered into another group. The 26 samples of S. japonica cv. jinhuai were clustered into two groups based on the phylogenetic tree constructed by matK sequences. The sample of seedling tree in Xianshui was clustered into one group, while the other 25 samples were clustered into another group. The clustering results of the rbcL sequence of S. japonica cv. jinhuai could not distinguish 28 resource samples. The phylogenetic tree constructed by the combined sequence of ITS2+psbA-trnH+rbcL+matK divided S. japonica cv. jinhuai resource samples into 4 groups. The 13 samples of seedling trees in Qiyang, Daoxian, Miaotou, Shaoshui, Shitang, Xianshui, Jiantang, and Xiangli, and grafting trees in Qiyang, Miaotou, Yongsui, Wenqiao, and Yangtang were clustered into one group. The sample of seedling tree in Wenqiao was clustered into one group. The sample of seedling tree in Daxu was clustered into one group. The remaining samples were clustered into another group.ConclusionPhylogenetic and mutation analysis provide the theoretic foundation to investigate the evolution of the resources of S. japonica cv. jinhuai, and evaluate their genuineness. The results of mutation points can be used to identify the related S. japonica cv. jinhuai resources. The findings of this study show that the combination of different gene sequences has an optimal effect on plant identification.
Abstract:ObjectiveThe law of fertilizer requirement serves as the basis for the fertilization of medicinal plants, development of special fertilizer, and high-quality medicinal materials.MethodThis study aims to explore the optimal potassium application rate for Panax ginseng to achieve high yield and quality of the medicinal material and targeted management of potassium fertilizer. To be specific, 6 concentration gradients (0, 2, 4, 8, 10, and 12 mmol·L-1) of potassium sulfate (potassium fertilizer) were designed and applied to the 4-year-old P. ginseng in CK, C1, C2, C3, C4, and C5 treatments, respectively. Thereby, the influence of potassium concentration on P. ginseng was observed.ResultWhen potassium sulfate was applied at 8 mmol·L-1, P. ginseng had the chlorophyll content of 32.13%, net photosynthetic rate of 2.548 8 µmol·m-2·s-1, and activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) significantly higher than those in the CK, C1, C4, and C5 treatments (P<0.05). The average fresh weight of P. ginseng roots was 6.25 g, 134% up from the CK, and the content of ginsenoside Rg1 (5.24 mg·g-1) and Re (4.17 mg·g-1) and total saponins (12.33 mg·g-1) was significantly higher than that in CK and other treatments (P<0.05). Thus, 8 mmol·L-1 potassium sulfate was most favorable for the growth and effective component accumulation of four-year-old P. ginseng.ConclusionThis study expounds the effect of potassium fertilizer on the yield and quality of P. ginseng, which is expected to help guide the precise application of potassium fertilizer in P. ginseng production in the field and lay a theoretical basis for the development of special fertilizer for P. ginseng and the optimization of fertilization technology.
Abstract:ObjectiveTo predict the potential targets and mechanism of Jingfang mixture in the treatment of H1N1 influenza and provide references for clinical application of Jingfang mixture.MethodThe active components and targets of Jingfang mixture against H1N1 influenza were screened out by Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),SwissTargetPrediction, and TargetNet. The targets of H1N1 influenza were obtained from GeneCards,Online Mendelian Inheritance in Man (OMIM), and DisGeNET and standardized by UniProt KB. The intersection targets were obtained by Venny 2.1.0. The "drug-component-target" network was constructed with Cytoscape 3.2.1 and analyzed for the topological attributes. The intersection targets were uploaded to STRING 11.5 to obtain the protein-protein interaction (PPI) network. Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were carried out by Metascape. Finally,the top active components ranked by degree were docked to the core targets by Autodock vina and visually analyzed by PyMOL. Balb/c female rats were used for experimental verification. Hematoxylin-eosin(HE) staining was used to observe the pathological changes in lung tissues. Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of tumor necrosis factor-α(TNF-α),interleukin-10(IL-10), and interleukin-17(IL-17). Real-time fluorescence-based quantitative polymerase chain reaction(Real-time PCR) and Western blot were used to detect the mRNA and protein expression levels in lung tissues.ResultThere were 144 active components in Jingfang mixture. A total of 421 target genes of Jingfang mixture and 2 956 targets of H1N1 influenza were identified,including 199 common targets. Topological analysis showed that the core components of Jingfang mixture against H1N1 influenza included quercetin,luteolin, and kaempferol,and the core targets included prostaglandin-endoperoxide synthase 2(PTGS2),estrogen receptor alpha(ESR1),inducible nitric oxide synthase 2(iNOS2),peroxisome proliferator-activated receptorγ(PPARγ),and cyclooxygenase-1(PTGS1). GO enrichment yielded 697 items in biological process (BP) (P<0.01), 59 items in molecular function (MF)(P<0.01), and 21 items in cellular component (CC) (P<0.01). A total of 132 signaling pathways (P<0.01) were obtained by KEGG enrichment analysis, including phosphatidylinositol 3-kinases(PI3K)/protein kinase B(Akt) signaling pathway and mitogen-activated protein kinase(MAPK) signaling pathway,most of which were related to the regulation of immune inflammation. Molecular docking showed that the binding energy of the active components of Jingfang mixture to the core targets was less than -5.0 kcal·mol-1,indicating good binding activity. HE staining showed that the lung tissues were significantly improved after drug intervention,and Real-time PCR and Western blot showed that Jingfang mixture could reduce the mRNA and protein expression of PI3K and Akt in lung tissues.ConclusionJingfang mixture can play an anti-viral effect against the influenza A virus through multiple components,multiple targets, and multiple pathways. The active components quercetin,luteolin, and kaempferol may control the inflammation and regulate immunity on the PI3K/Akt,MAPK, and other signaling pathways by acting on targets such as PTGS2,ESR1,iNOS2,PPARγ, and PTGS1.
Abstract:ObjectiveTo explore the potential mechanism of Polygonati Rhizoma on the treatment of osteoporosis (OP) based on network pharmacology and molecular docking method and to verify the mechanism by experiments.MethodThe main active ingredients and corresponding targets of Polygonati Rhizoma were screened out from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP) 2.3 by conditional searching. The treatment targets were obtained from the genes related to OP and DisGeNET 7.0. The potential target genes of Polygonati Rhizoma for treating OP were obtained by the crossing of the corresponding targets and the treatment targets. Cytoscape 3.7.1 was used to construct the “Polygonati Rhizoma-active ingredient-potential target” network. The protein-protein interaction (PPI) analysis was carried out by STRING 11.0, and the PPI network was constructed. Metascape 3.5 was used to conduct Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of the key targets. The core ingredients and key targets of Polygonati Rhizoma were selected for molecular docking by AutoDock Vina 1.1.2. Finally, the effect of β-sitosterol on osteogenic differentiation of MC3T3-E1 cells in rats was observed.ResultTwelve active ingredients and 32 potential targets of Polygonati Rhizoma for OP treatment were screened out. Six active ingredients including baicalein and β-sitosterol and key targets including protein kinase 1 (Akt1), tumor suppressor p53 (TP53), vascular endothelial growth factorA (VEGFA), proto-oncogene Jun (JUN), matrix metalloproteinase-9 (MMP-9), and proto-oncogene c-Fos (FOS) were obtained by Cytoscape 3.7.1 topological analysis. A total of 995 GO entries and 181 signaling pathways involving the response to reactive oxygen species and regulations of growth were obtained from GO and KEGG enrichment analyses. The results of molecular docking showed that the core active ingredients possessed good binding activities with the respective key targets. The results of cell experiments showed that β-sitosterol promoted the osteogenic differentiation at the concentration of 2.5 μmol·L-1 and 5 μmol·L-1.ConclusionPolygonati Rhizoma had the therapeutic effect on treating OP by regulating inflammation, oxidative stress, apoptosis, and metabolism. The β-sitosterol significantly promoted the osteogenic differentiation of MC3T3-E1 cells.
Abstract:ObjectiveTo explore the anti-testicular inflammation mechanism of Wuzi Yanzongwan through network pharmacological analysis and experimental verification.MethodThe active ingredients of Wuzi Yanzongwan were searched in the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), and their targets were predicted via SwissTargetPredicition. GeneCards and Online Mendelian Inheritance in Man (OMIM) were used to obtain the known targets for the treatment of testicular inflammation. STRING 11.0 was employed to construct the protein-protein interaction (PPI) network, and Cytoscape 3.7.1 to construct the “herbal medicine-active ingredient-target” network of Wuzi Yanzongwan and “active ingredient-disease target” network. Metascape was used for gene ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment of signaling pathways.ResultA total of 72 active ingredients and 624 potential targets of Wuzi Yanzongwan were screened out, and 196 key targets were identified for the treatment of testicular inflammation by Wuzi Yanzongwan. The results of GO annotation showed that the anti-testicular inflammation targets of Wuzi Yanzongwan were involved in the biological processes such as the response to molecule of bacterial origin, the positive regulation of response to external stimulus, and the response to extracellular stimulus. KEGG pathway enrichment revealed that the major pathways associated with the treatment (P<0.01) included the pathways in cancer, epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor resistance, transcriptional misregulation in cancer, calcium signaling pathway, apelin signaling pathway, and NOD-like receptor signaling pathway. Wuzi Yanzongwan may exert the anti-testicular inflammation effect by inhibiting the activation of the NLR family pyrin domain-containing 3 (NLRP3) inflammasome pathway to alleviate the inflammatory response, which verifies the prediction results based on network pharmacology to a certain extent.ConclusionThe inhibitory effect of Wuzi Yanzongwan on testicular inflammation is multi-target, multi-pathway, and multi-mechanism. The findings of this study provide evidence support for the clinical application of Wuzi Yanzongwan.
Abstract:The present study explored the effective approaches to realize the leading role of traditional Chinese medicine (TCM) in preventing diseases, the synergistic role in treating serious diseases, the core role in the rehabilitation of diseases and summarized the experience to provide feasible plans for the evaluation of other dominant diseases of TCM. To evaluate the effectiveness, safety, and economy of TCM in the treatment of ischemic stroke, encephalopathy project team of the China Center for Evidence-based Traditional Chinese Medicine(CCEBTCM) established an evaluation group to determine the work plan and complete the evaluation work. The concepts of the evaluation involved high-quality evidence, expert opinion survey, expert interview, and drug catalog. Under the guidance of clinical experts and methodologists, the evaluation work was completed in accordance with four steps, i.e., plan making, data collection and data extraction, evidence synthesis and evaluation, and report writing with the rapid review method. Through the review of TCM and western medicine experts, the advantage of TCM in the treatment of ischemic stroke was positioned in the convalescence period with the predominant effects of improving the neurological function defect and improving the daily living ability. In the convalescence period of stroke, TCM treatment could improve post-stroke motor dysfunction, post-stroke cognitive impairment, consciousness disorder, swallowing disorder, aphasia, constipation, urinary function, diplopia, etc., and the advantages of acupuncture, Chinese medicine, and traditional exercise were more prominent. In terms of safety, TCM treatment of ischemic stroke showed lower incidence of adverse reactions, fewer adverse events, and a milder degree of related symptoms. In terms of economic performance, the combined treatment of TCM and western medicine played a synergistic role and made the treatment cost more reasonable. Compared with conventional intervention, the integrated TCM and western medicine rehabilitation program showed more economic and social benefits.
Keywords:dominant disease of traditional Chinese medicine;ischemic stroke;positioning of advantage;rapid review;evidence-based evidence
Abstract:Chronic heart failure is a serious heart disease with dyspnea and limited activity tolerance as the main clinical manifestations. Autophagy is a self-protection mechanism in eukaryotic cells and plays an important role in the development of heart failure. Appropriately increasing the level of autophagy during the compensated stage of heart failure and timely removal of necrotic myocardial organelles and other harmful garbage can inhibit myocardial hypertrophy to a certain extent,alleviate myocardial remodeling,and delay heart failure. The theory of healthy Qi and pathogenic Qi is an important basic theory for explaining the occurrence of diseases,and struggle between healthy Qi and pathogenic Qi exists in the entire onset of chronic heart failure,which may lead to pathogenic Qi invasion and healthy Qi deficiency. The regulatory effect of autophagy on cardiomyocytes is similar to the theory of healthy Qi and pathogenic Qi in traditional Chinese medicine (TCM). Autophagy is the body's self-regulatory mechanism for healthy Qi and pathogenic Qi in a dose-effect manner,Specifically,autophagy can only protect the body's cells to a certain extent,and healthy Qi can only take effect within a certain range and degree. To protect the body from external pathogenic factors,excessive or insufficient autophagy may destroy the stability of the body's environment. In this regard,we use the theory of healthy Qi and pathogenic Qi as a starting point to clarify the function of autophagy in the development of chronic heart failure from a macro and micro perspective,and propose adjusting the balance of healthy Qi and pathogenic Qi in the body to regulate the autophagy of cardiomyocytes. The principle of prevention and treatment is expected to lay the foundation for modern research on the function of autophagy in the development of chronic heart failure in TCM,find novel therapy for chronic heart failure at different stages,and provide new insights into the diagnosis and treatment of chronic heart failure.
Keywords:theory of healthy Qi and pathogenic Qi;struggle between healthy Qi and pathogenic Qi;balance between healthy Qi and pathogenic Qi;autophagy;chronic heart failure
Abstract:Bronchial asthma, a chronic inflammatory airway disease, belongs to the category of wheezing disease in the system of traditional Chinese medicine (TCM). The wheezing symptom of this disease is mainly caused by the imbalance of lung Qi. According to the theory of five flavor compatibility, the Chinese medicinal materials with five different flavors (pungent, bitter, sour, sweet, and salty) can be combined to produce new functions. The pungent medicinal materials have dispersing effect and the bitter medicinal materials have discharging effect, which are important components in the theory of five flavor compatibility. Pungent herbs and bitter herbs can relieve the adverse lung Qi, occupying an important position in the current medication for the treatment of asthma. However, there is still a lack of in-depth analysis of the TCM theory and mechanism of the compatibility of pungent herbs and bitter herbs in the treatment of asthma. The molecular mechanisms of action of pungent herbs and bitter herbs are closely related to transient receptor potential (TRP) channels and bitter taste receptors (TAS2Rs), respectively. Ca2+ signaling has been recognized in the process of asthma and is involved in the development of multiple symptoms of asthma. The TRP channels and TAS2Rs located on the cell membrane have been proved to directly regulate the intracellular Ca2+ signal and play a role in the treatment of asthma. Therefore, the dispersing effect of pungent herbs and the discharging effect of bitter herbs may be realized through the Ca2+ signaling pathway mediated by TRPs/TAS2Rs. We summarized the theoretical understanding and modern studies of pungent herbs dispersing lung Qi and bitter herbs discharging lung Qi, aiming to explain the internal relationship and mechanism of the compatibility of pungent herbs and bitter herbs in the treatment of asthma from the perspective of TCM theory and modern medicine. The compatibility of pungent herbs and bitter herbs based on the theory of five flavor compatibility for the treatment of asthma has a solid theoretical basis of TCM, and its mechanism can be verified by modern research. Therefore, it may be a main research direction in the treatment of asthma by Chinese medicinal herbs in the future.
Keywords:five flavor compatibility;dispersing effect of pungent herbs and discharging effect of bitter herbs;transient receptor potential (TRP) channels;bitter taste receptors (TAS2Rs);asthma
Abstract:Traditional Chinese medicine (TCM), which owns abundant chemical components and complex action pathways, has been widely recognized in the prevention and treatment of diseases. Some analysis methods have been emerged in order to ensure the quality of TCM and to develop new TCM drugs. Matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS) is a soft ionization mass spectrometric technique with the advantages of high throughput, high sensitivity, low cost and so on. It provides technical support for the molecular level study on TCM. At present, this technique has been used in the field of composition analysis and metabonomics research of TCM, and plays an important role in the identification of Chinese herbal medicines, real-time molecular screening and the construction of metabolic network pathway of active ingredients. Among them, the selection of appropriate matrix and sample preparation technology is the key to ensure the detection effect of MALDI-MS. With the development and optimization of new matrix, the continuous improvement of sample preparation technology and the combination of MALDI-MS with various analytical methods will greatly improve the detection effect. Based on this, this paper discusses the application of MALDI-MS in TCM, including high-throughput detection of active ingredients in TCM, monitoring of the original medicines and their metabolites in vivo, and in situ visualization and characterization of tissue distribution information of active ingredients in TCM. It also discusses the application prospect and existing problems of MALDI-MS in TCM, so as to provide technical support for the identification of active ingredients in TCM, drug utilization and metabolism.
Keywords:matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS);traditional Chinese medicine;active ingredients;metabolites;mass spectrometry imaging;in situ visualization;tissue distribution
Abstract:Diabetes is a metabolic disease mainly characterized by hyperglycemia due to inadequate insulin secretion. And persistent hyperglycemia can cause chronic damage or dysfunction of eyes, kidneys, heart, blood vessels and nerves. Polysaccharides are high molecular carbohydrates polymerized by glycosidic bonds from more than 10 monosaccharide molecules of the same or different types. They have the advantages of wide sources, high safety and low toxic and so on. As one of the important effective components of traditional Chinese medicine, polysaccharides have biological activities such as immune regulation, anti-oxidation, anti-tumor, lowering blood sugar and so on. The structure is directly related to biological activities, and the advanced structure of polysaccharides is based on the primary structure. Exploring the primary structure of polysaccharides is the key task of lowering blood sugar and improving diabetic complications. This paper summarizes the monosaccharide composition of the primary structure of Chinese medicine polysaccharides, and the mechanism of Chinese medicine polysaccharides improving diabetes is emphasized by increasing the secretion and release of insulin, increasing the islet β cell number, upregulating insulin receptor level, improving glucose and lipid metabolism, inhibiting inflammatory response, improving oxidative stress and regulating phosphatidylinositol-3-kinase(PI3K)/protein kinase B (Akt), mitogen activated protein kinase, cyclic adenosine monophosphate(cAMP)/protein kinase A(PKA) and adenosine monophosphate activated protein kinase(AMPK) signaling pathways. At the same time, we also summarized the prevention and treatment of Chinese medicine polysaccharides in diabetic nephropathy, diabetic cardiomyopathy, diabetic ophthalmopathy and diabetic peripheral neuropathy, in order to provide a theoretical basis for new drug development and clinical application of Chinese medicine polysaccharides in the intervention of diabetes and its complications.
Keywords:Chinese medicine polysaccharide;diabetes;diabetic complications;diabetic nephropathy;diabetic cardiomyopathy;mechanism of action;structure
Abstract:Esophageal cancer is a digestive tract malignancy with high morbidity and mortality and mainly occurs in males. The 5-year survival rate is lower than 20%. In China, the morbidity and mortality of esophageal cancer rank the first in the world, seriously threatening national health. The pathogenesis of esophageal cancer is diverse, which is generally considered as the consequence of environmental-genetic-gene interaction. In addition to genetic factors and regional characteristics, gene mutation, RNA interference, DNA damage repair, tumor microenvironment, dietary habit, chronic adverse stimulation, and inflammatory reaction are all involved in the occurrence and development of esophageal cancer. However, there is no unified and accurate conclusion. Clarifying the exact pathogenesis of esophageal cancer is of great significance for its early screening, diagnosis, prevention, treatment, and prognosis. Surgery, radiotherapy, and chemotherapy are the three effective methods for the treatment of esophageal cancer. However, due to the atypical early symptoms, most patients have missed the best operation period when diagnosed, resulting in poor clinical prognosis. Moreover, radiotherapy and chemotherapy will cause side effects such as loss of appetite, low immune function, esophagitis, pneumonia, and malnutrition, which is not conducive to the prognosis and treatment maintenance of patients. With definite efficacies on esophageal cancer, traditional Chinese medicine (TCM), which is flexible and diverse in the treatment, can primarily or alternatively be involved in the treatment of esophageal cancer. TCM can eliminate postoperative complications and postoperative infections and relieve adverse gastrointestinal reactions, weakened immune function, and organ damage caused by radiotherapy and chemotherapy. It can enhance clinical efficacy and improve the quality of life of patients. Therefore, it is necessary to systematically summarize the clear pathogenesis or risk factors of esophageal cancer and review the clinical characteristics of TCM in the prevention and treatment of esophageal cancer to facilitate the early screening, diagnosis, and treatment of esophageal cancer and promote the application of TCM in the prevention and treatment of esophageal cancer and related adverse reactions.
Keywords:esophageal cancer;pathogenesis;traditional Chinese medicine;action characteristics
Abstract:Atopic dermatitis is a chronic, refractory and inflammatory skin disease with the clinical manifestations of severe pruritus and recurrent attacks. It has a high incidence and is closely correlated with other allergic, autoimmune or infectious diseases, which can cause a variety of secondary diseases and mental and psychological disorders, seriously affecting the life quality of patients. Chinese herbal medicines have been used for the prevention and treatment of atopic dermatitis for thousands of years, and many Chinese herbal medicines (including compound prescriptions) effective for this disease have been recorded. These medicines generally have little adverse reactions and the treated patients have low recurrence rate of atopic dermatitis. According to the evidence of modern medicine, the onset of atopic dermatitis is related to the impairment of skin barrier function, abnormal immune response, and abnormal differentiation of mast cells, antigen-presenting cells, and eosinophils. Additionally, it is associated with mental, endocrine, metabolic and other factors. The defect of skin barrier function and the dysfunction of immune system are the main pathogenesis of atopic dermatitis. In recent years, scientists have achieved certain progress in improving skin barrier function with Chinese herbal medicines. This paper systematically summarizes the studies about the application of Chinese herbal medicines in regulating the expression of epidermal proteins, epidermal lipids, aquaporins, tight junction proteins, and antimicrobial peptides in recent 10 years, aiming to clarify the pathological mechanism and provide reference for the clinical research and application of Chinese herbal medicines in the treatment of atopic dermatitis.