Abstract:ObjectiveTo establish a method for evaluating the biological activity of water extract lyophilized powder of Qingjin Huatantang based on the phagocytic and secretory functions of macrophages, and to control the quality of this formula from the biological activity level.MethodThe phagocytic and inflammation models of RAW264.7 macrophages were established, the inhibition rates of water extract lyophilized powder of Qingjin Huatantang on interleukin-6 (IL-6) secretion and phagocytic index of neutral red of RAW264.7 macrophages were chosen as indicators to investigate the biological activity of Qingjin Huatantang, and the biological limit was searched.ResultThe optimal inoculation density of RAW264.7 macrophages was 3×105 pcs/mL, and the concentration of lipopolysaccharide (LPS) was 1 mg·L-1 after treatment for 24 h. When the concentration was 500 mg·L-1, water extract lyophilized powder of Qingjin Huatantang had no toxicity and no obvious promotion effect on the proliferation of RAW264.7 macrophages, and at this concentration, the phagocytosis of RAW264.7 macrophages for neutral red was significantly promoted, the phagocytic index was >113%. In addition, the lyophilized powder had a significant and stable inhibitory effect on IL-6 secretion of RAW264.7 macrophages induced by LPS, the inhibitory rate was >45%.ConclusionCombined with the anti-inflammatory and immunomodulatory effects of Qingjin Huatantang, this study establishes an in vitro biological limit method for evaluating the quality of water extract of Qingjin Huatantang based on the phagocytic and secretory functions of RAW264.7 macrophages, and 500 mg·L-1 was confirmed as the limit concentration. Under the limit concentration, Qingjin Huatantang water extract can significantly promote the phagocytic index of macrophages or significantly inhibit the secretion of IL-6 of RAW264.7 macrophages induced by LPS, which can be judged as qualified.
Abstract:ObjectiveTo observe the effect of Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma, and Chuanxiong Rhizoma extract (GNC) on mitochondrial oxidative stress in hydrogen peroxide (H2O2)-induced aging of human umbilical vein endothelial cells (HUVECs), and explore the therapeutic mechanism of GNC on aging HUVECs.MethodThe HUVECs were classified into the control group (control), H2O2 model group (H2O2), H2O2 + DMSO group (DMSO, 1 mL·L-1), resveratrol group (Resv, 8 μmol·L-1), and low- (200 mg·L-1), medium- (300 mg·L-1), and high-dose (400 mg·L-1) GNC (GNC-L, GNC-M, and GNC-H) groups. Except control group and H2O2 group, the other groups were intervened with corresponding agents. Subsequently, 300 μmol·L-1 H2O2 was given to other groups except the control group for 4 h to induce aging, and then the cells were cultured in normal media for 24 h. The aging degree, cell cycle, and mitochondrial reactive oxygen species (mtROS) level were determined by SA-β-galactosidase (SA-β-Gal) staining, flow cytometry, and MitoSox red fluorescence staining, respectively. JC-10 was used as a fluorescent probe to detect the changes in mitochondrial membrane potential, and Western blot was performed to detect the expression of manganese superoxide dismutase (MnSOD) and p-p66 proteins.ResultThe SA-β-gal staining results showed that H2O2 group had increased blue-stained cells compared with other groups (P<0.01). Compared with those in the control group, the ratio of G0/G1 phase cells significantly increased (P<0.05) and that of G2/M phase cells decreased (P<0.05) in the H2O2 group. Compared with those in the H2O2 group, the proportion of G0/G1 cells decreased (P<0.05) while that of G2/M cells increased (P<0.05) in GNC-H groups and Resv group. The fluorescence staining for determining mitochondrial ROS level showed that the H2O2 group had weakened fluorescence intensity than the control, GNC-H, and GNC-M groups (P<0.05). The mitochondrial membrane potential fluorescence intensity of the H2O2 group was weaker than that of the control, GNC-H, GNC-M, and GNC-L groups (P<0.01), as well as the Resv group (P<0.05). Western blot showed that the protein level of MnSOD was significantly lower in the H2O2 group than in the control, GNS-H, and GNS-M groups (P<0.05), whereas the protein level of p-p66 showed an opposite trend (P<0.01), indicating that the medication can alleviate the intracellular mitochondrial oxidative stress.ConclusionGNC can delay the H2O2-induced aging of vascular endothelial cells. The GNC intervention significantly regulated the mitochondrial ROS, mitochondrial membrane potential, and related proteins MnSOD and p-p66 to alleviate oxidative stress. Chinese medicinal materials may delay the aging of vascular endothelial cells by inhibiting mitochondrial oxidative stress.
Keywords:aging;vascular aging;aging of endothelial cells;Ginseng Radix et Rhizoma, Notoginseng Radix et Rhizoma, and Chuanxiong Rhizoma extract;oxidative stress
Abstract:Depression is a threat to human health due to high incidence, recurrence, and disability rates. At present, the complex etiology and pathogenesis of depression are still unclear, and such hypotheses as monoamine neurotransmitters and their receptor abnormality, hyperactivity of hypothalamic-pituitary-adrenal (HPA) axis, inflammation, and neuron damage and remodeling have been put forward. Anti-depressants developed based on the pathogenesis have a certain effect, but there also exist some problems like low cure rate, poor compliance, relapse after discontinuation, and obvious side effects. According to Zhongjing's theory, depression falls into the categories of depression syndrome, visceral agitation, insomnia, and lily disease, and it is mainly located in the liver, involving the spleen, heart, and kidney. The pathogenesis mainly lies in qi stagnation and zang-fu organ dysfunction. Attention should be focused on regulating qi and relieving depression. Zhongjing's antidepressant prescriptions have exhibited good clinical efficacy in the treatment of depression, reflecting the multi-pathway action advantages of Chinese herbs. Based on the pathogenesis of depression and domestic and foreign literature on the intervention of depression with Zhongjing's prescriptions available in the past 20 years, this paper summarized the mechanisms of Zhongjing's prescriptions against depression from the experimental and clinical research aspects, in order to provide reference for clinical treatment of depression with Zhongjing's prescriptions.
Abstract:ObjectiveTo investigate the effect and mechanism of Chaihu Jia Longgu Mulitang (CJLM) on hippocampal NOD-like receptor protein 3 (NLRP3)inflammasome pathway in rats with depression.MethodSixty male SD rats were randomly divided into a normal group,a model group, a MCC950 (1 mg·kg-1) group, and high- (13 g·kg-1), medium- (6.5 g·kg-1), and low-dose (3.25 g·kg-1) Chaihu Jia Longgu Mulitang groups, with 10 rats in each group.The depression model was induced by isolation combined with chronic unpredictable mild stimulation(CUMS) in rats except for those in the normal group. Rats were treated correspondingly for 21 days by intraperitoneal injection in the MCC950 group and gavage in other groups. The normal group and the model group received an equal volume of normal saline. The depression-like behaviors of rats were observed by sucrose preference test (SPT) and novelty-suppressed feeding test. Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of interleukin-1β (IL-1β) and IL-18 in the hippocampus of depressed rats. Western blot was used to detect the protein levels of NLRP3, apoptosis-associated speck-like protein containing a C-terminal caspase recruitment domain (ASC), and Caspase-1.ResultCompared with the normal group, the model group showed decreased sucrose preference rate (P<0.01), prolonged novelty-suppressed feeding time (P<0.01), enhanced protein expression of NLRP3,ASC, and caspase-1 (P<0.05, P<0.01), and elevated expression of IL-1β and IL-18 (P<0.01).ConclusionCJLM can alleviate depression-like behaviors in CUMS-induced model rats, and the underlying mechanism is related to the inhibition of the NLRP3 inflammasome pathway.
Keywords:depression;Chaihu Jia Longgu Mulitang;inflammation;hippocampus;NOD-like receptor protein 3(NLRP3)inflammasome
Abstract:ObjectiveTo observe the effect of Sinisan on the brain-derived neurotrophic factor (BDNF)/tyrosine kinase receptor B (TrKB), 5-hydroxytryptamine (5-HT)/5-HT1A receptor (5-HT1AR), and hypothalamus-pituitary-adrenal (HPA) axis in depressed rats, and explore the antidepressant mechanism of Sinisan based on BDNF/TrKB, 5-HT/5-HT1AR, and HPA axis.MethodA total of 120 male Wistar rats were randomly divided into a normal group, a model group, a fluoxetine (0.01 g·kg-1) group, and low- (1.25 g·kg-1), medium- (2.5 g·kg-1), and high-dose (5 g·kg-1) Sinisan groups, with 20 rats in each group. The depression model was induced by isolation combined with chronic unpredictable mild stimulation(CUMS) in rats except for those in the normal group for 21 days. Rats were then treated correspondingly once a day for 21 days by gavage. Those in the normal group and the model group received an equal volume of normal saline. During the intervention, the model rats were stimulated continuously. The depressive state of CUMS model rats was evaluated by sucrose preference test and open field test. Enzyme-linked immunosorbent assay (ELISA) was used to determine the levels of corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH), and corticosterone (CORT) in the plasma and BDNF and 5-HT levels in the hippocampal homogenate. The mRNA expression of hippocampal TrKB, 5-HT1AR, glucocorticoid receptor (GR), and mineralocorticoid receptor (MR) was detected by real-time fluorescence-based quantitative polymerase chain reaction (Real-time PCR). The protein expression of hippocampal TrKB, 5-HT1AR, GR, and MR was detected by Western blot. The histomorphological changes of the hippocampus were observed by hematoxylin-eosin (HE) staining.ResultCompared with the normal group, the model group showed decreased sucrose preference rate (P<0.01), reduced horizontal and vertical scores in the open field test (P<0.01), increased plasma content of CRH, ACTH, and CORT (P<0.01), declining content of BDNF and 5-HT in the hippocampus (P<0.01), dwindled mRNA and protein expression levels of TrKB, 5-HT1AR, and GR (P<0.01), elevated mRNA and protein expression of MR (P<0.01), and damaged hippocampal neurons revealed by HE staining. Compared with the model group, the groups with drug intervention showed increased sucrose preference rate (P<0.01) and horizontal and vertical scores in the open field test (P<0.05, P<0.01), decreased content of plasma CRH, ACTH, and CORT (P<0.05, P<0.01), elevated content of hippocampal BDNF and 5-HT (P<0.05, P<0.01), elevated mRNA and protein expression levels of hippocampal TrKB, 5-HT1AR, and GR (P<0.05, P<0.01), reduced mRNA and protein expression levels of hippocampal MR (P<0.05, P<0.01), and recovered hippocampal neurons as revealed by HE staining.ConclusionSinisan can exert a significant antidepressant effect by increasing hippocampal BDNF and 5-HT content, up-regulating TrKB, 5-HT1AR, and GR mRNA and protein expression, down-regulating MR mRNA and protein expression, inhibiting HPA axis hypertrophy, and enhancing the regeneration and repair of hippocampal neurons.
Abstract:ObjectiveTo observe the clinical therapeutic effect of Suanzaoren Tang combined with fluoxetine in the treatment of patients with depression of liver stagnation and blood deficiency accompanied by insomnia.MethodThe patients with depression of liver stagnation and blood deficiency accompanied by insomnia (120 cases) were randomly divided into an observation group and a control group, with 60 cases in each group. The patients in the observation group received Suanzaoren Tang combined with fluoxetine, and those in the control group received fluoxetine. The course of treatment was eight weeks. The clinical efficacy was evaluated with Hamilton Depression Rating Scale (HAMD), Pittsburgh Sleep Quality Index(PSQI), and Activities of Daily Living (ADL) score. Enzyme-linked immunosorbent assay (ELISA) was used to determine the plasma levels of 5-hydroxytryptamine (5-HT), norepinephrine (NE),brain-derived neurotrophic factor (BDNF), glial cell-derived neurotrophic factor (GDNF), neuron-specific enolase (NSE), and S100β.ResultAfter eight weeks of treatment, the scores of HAMD and PSQI were reduced(P<0.01), while the scores of ADL were elevated(P<0.01),and the levels of 5-HT, NE, GDNF and BDNF were up-regulated (P<0.01) in the plasma of patients in the observation group as compared with those before treatment. After treatment, compared with the control group, the observation group showed increased total effective rate(P<0.01), decreased scores of HAMD and PSQI (P<0.01), elevated score of ADL(P<0.01), up-regulated levels of 5-HT, NE, GDNF and BDNF in plasma, and declining NSE and S100β(P<0.01).ConclusionSuanzaoren Tang combined with fluoxetine is superior to fluoxetine alone in treating the depression of liver stagnation and blood deficiency accompanied by insomnia. Its therapeutic effect is achieved by increasing the release of monoamine neurotransmitters and promoting the secretion of BDNF and GDNF in the brain.
Abstract:ObjectiveTo observe the effects of modified Liuwei Dihuangtang on serum fibroblast growth factor 23 (FGF23), full-length intact parathyroid hormone (iPTH), and 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] levels and Klotho and FGF23 protein expression in renal and bone tissues of rats exposed to high phosphorus combined with adenine, so as to explore the mechanism of modified Liuwei Dihuangtang against renal osteopathy.MethodOne hundred and thirty healthy adult SD rats were randomly divided into five groups, namely normal group(n=10),high phosphorus group(n=30),model group(n=30),modified Liuwei Dihuangtang group(n=30) , and calcitriol group(n=30),and rats in each group were further classified based on three time points, namely 8,10, and 12 weeks. Rats in the normal group were fed with normal diet, the ones in the high phosphorus group with high phosphorus diet, and those in the other groups with adenine and high phosphorus diet for inducing renal osteopathy. Rats in the normal group,high phosphorus group, and model group were intragastrically administered with distilled water (10 mL·kg-1·d-1),the ones in the modified Liuwei Dihuangtang group with modified Liuwei Dihuangtang (2.556 g·kg-1·d-1) , and those in the calcitriol group with calcitriol (0.09 μg·kg-1·d-1).ResultCompared with the normal group and high phosphorus group at the weeks of 8,10 and 12,the model group displayed significantly elevated blood urea nitrogen(BUN),serum creatinine(SCr),serum phosphorus,iPTH,FGF23,renal interstitial fibrosis score, and FGF23 expression in renal and bone tissues, but lowered serum calcium and 1,25(OH)2D3 and Klotho protein expression in renal and bone tissues(P<0.05 ,P<0.01). Compared with the model group at the weeks of 8,10 and 12, the modified Liuwei Dihuangtang and calcitriol both significantly decreased the serum BUN,SCr,serum phosphorus,iPTH, FGF23, tubulointerstitial semi-quantitative score, and FGF23 expression in renal and bone tissues, while increased the serum calcium,1,25(OH)2D3, and Klotho protein expression in renal and bone tissues (P<0.05,P<0.01). There was no significant difference in the above-mentioned indexes between the modified Liuwei Dihuangtang group and the calcitriol group at the same time point.ConclusionKlotho-FGF23 axis is probably involved in renal osteopathy. The modified Liuwei Dihuangtang effectively improves renal function,alleviates pathological changes in renal and bone tissues,and regulates calcium and phosphorus metabolism to protect the bone, which is related to its regulation of Klotho-FGF23 axis.
Keywords:modified Liuwei Dihuangtang;renal osteopathy;fibroblast growth factor 23 (FGF23);Klotho protein
Abstract:ObjectiveTo investigate the effect and mechanism of Fuzheng Touxie prescription (FZTX) on the immune homeostasis of drug-resistant Pseudomonas aeruginosa lung infection in rats at different time points.MethodA total of 168 rats were divided into a blank group (n=8),a model group (n=40),a Touxie (TX) group (n=40),an early Fuzheng (FZ) group (n=40), and a delayed FZ group (n=40). The blank group was given distilled water by gavage, the model group was given distilled water by gavage after infection,the TX group was given clear heat and penetrate evil drug free decoction granules(3.5 g·kg-1) by gavage after infection, the early FZ group was given Fuzheng Touxie whole formula free decoction granules(10.75 g·kg-1) by gavage after infection, the delayed FZ group was given clear heat and penetrate evil drug free decoction granules by gavage after infection, on the third day plus Fuzheng drug free decoction granules[(3.5+10.75) g·kg-1] by gavage, the three treatment groups were gavaged twice a day, 2 mL each time .Each drug treatment group was divided into five groups according to five time points (3 h,1 d,3 d,5 d, and 7 d), with eight rats in each group. The levels of tumor necrosis factor-α(TNF-α),high mobility group protein 1(HMGB1),interleukin-10(IL-10), and tumor necrosis factor -α-induced protein-8-like2 (TIPE2) were measured by enzyme-linked immunosorbent assay (ELISA), and HMGB1 protein expression level by Western blot.ResultAt 3 h,the TNF-α content in the drug treatment groups was higher than that in the blank group and the model group (P<0.05). At 3 d,the TNF-α content in the early FZ group and the delayed FZ group was lower than that in the model group (P<0.05) and the TX group (P<0.05). At 1 d,the HMGB1 content in the TX group and the delayed FZ group was higher than that in the model group (P<0.05). At 5 d,the HMGB1 content was lower in the delayed FZ group than in the model group (P<0.05). At 7 d,HMGB1 protein expression in the model group was higher than that in the blank group (P<0.05) and the early FZ group (P<0.05). At 3 d,the IL-10 content was significantly higher in both the early FZ group and the delayed FZ group than that in the model group (P<0.05). At 5 d,the IL-10 content was higher in the early FZ group than that in the TX group (P<0.05). At 7 d,the IL-10 content in the early FZ group and the delayed FZ group was lower than that in the TX group (P<0.05). At 5 d,the TIPE2 content in the early FZ group was lower than that in the model group (P<0.05). At 7 d,the TIPE2 content in the TX group and the delayed FZ group was lower than that in the model group (P<0.05).ConclusionFZTX or modified prescription can promote the inflammatory response to eliminate pathogenic bacteria in the early stage and suppress the inflammatory response in the late stage to avoid the inflammatory cascade effect and lung tissue damage,indicating that Fuzheng drugs have an important role in maintaining the immune homeostasis of the body after infection.
Keywords:drug-resistant Pseudomonas aeruginosa;lung infection model rats;immune homeostasis;Fuzheng Touxie prescription
Abstract:ObjectiveTo observe effect of Jingulian capsule on the proliferation of human breast cancer MDA-MB-231 cells and investigate its action mechanism against triple negative breast cancer (TNBC).MethodThe ingredients of Jingulian capsule were identified by ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS). The inhibitory effect of Jingulian capsule at different doses (0.125,0.25,0.5,1,and 2 g·L-1) against the proliferation of MDA-MB-231 cells were detected by methyl thiazolyl tetrazolium (MTT) assay. After treatment for 24 h, the morphological changes in nuclear apoptosis of MDA-MB-231 cells were detected by Hoechst 33258 staining. The effect of different concentrations of Jingulian capsule on the apoptosis and cycle of MDA-MB-231 cells after different treatment time were determined by flow cytometry. The protein expression levels of Poly-ADP-ribose polymeras (PARP), proto-oncogene c-Myc, cyclin B1, and phosphorylated extracellular signal-regulated kinase (p-ERK) in each group were assayed by Western blot.ResultA total of 113 compounds in Jingulian capsule were identified by UPLC-MS/MS. As revealed by MTT assay,compared with blank group,Jingulian capsule (0.125,0.25,0.5,1,2 g·L-1) significantly inhibited viability of MDA-MB-231 cells (P<0.01), with the half maximal inhibitory concentration ( IC50) of(0.13±0.02)g·L-1. According to flow cytometry,compared with the blank group,Jingulian capsule at 1 g·L-1 significantly induced the apoptosis of MDA-MB-231 cells (P<0.05)and Jingulian capsule at 0.5, 1 g·L-1 obviously increased the number of MDA-MB-231 cells in S phase (P<0.05,P<0.01). The results of Western blotting demonstrated that the protein expression levels of PARP,c-Myc,and cyclin B1 in 0.5, 1 g·L-1 Jingulian capsule groups were remarkably down-regulated as compared with those in the blank group(P<0.01), and the protein expression level of p-ERK in 1 g·L-1 Jingulian capsule group was also down-regulated (P<0.01).ConclusionJingulian capsule is able to inhibit the proliferation of MDA-MB-231 cells,induce S phase cell cycle arrest, and promote their apoptosis, which may be related to the inactivation of the MAPK signaling pathway.
Abstract:ObjectiveTo investigate the effect of Huoxue Jiedu Runzao prescription on the morphology, apoptosis, and function of submandibular gland in the mouse model of Sjögren's syndrome (SS) and its functioning mechanism, we analyzed the expression of the apoptosis inhibitor Survivin in the submandibular gland cells of SS mice.MethodFemale BALB/c57 mice were selected as the normal group. The naive non-obese diabetic (NOD/Ltj) female mice were selected as the SS model, which were randomly assigned into the model group, Paeoniae Radix Alba total glucosides capsule (0.234 g·kg-1) group, and low-, medium-, and high-dose (15.6, 31.2, 62.4 g·kg-1, respectively) Huoxue Jiedu Runzao prescription groups. Each group had 15 mice. The morphological and functional changes of submandibular gland and the Survivin expression were observed and measured after 8 weeks of drug intervention. Survivin expression was determined by immunohistochemistry (IHC), Western blot, and reverse transcription-polymerase chain reaction (RT-PCR).ResultCompared with normal group, salivary flow and submandibular gland index in model group were significantly decreased (P<0.01), and histopathological score of submandibular gland was significantly increased (P<0.01). Western blot showed that Survivin protein expression was significantly decreased (P<0.05). IHC showed that, Survivin mRNA expression was significantly decreased (P<0.01), and RT-PCR results showed that Survivin mRNA expression was significantly decreased (P<0.01). Compared with model group, salivary flow and submandibular gland index of mice in Huoxue Jiudu Runzao prescription groups and Paeoniae Radix Alba total glucosides capsules groups were significantly increased (P<0.05), histopathological score of submandibular gland was significantly decreased (P<0.05), IHC results showed that Survivin expression was significantly increased (P<0.01). Western blot and RT-PCR results showed that Survivin protein and mRNA expression of Huoxue Jiudu Runzao prescription high-dose group and Paeoniae Radix Alba total glucosides capsule group were significantly increased (P<0.05).ConclusionHuoxue Jiedu Runzao prescription can improve the secretion function of submandibular acinus, increase the submandibular gland index, and saliva secretion of SS mice by up-regulating survivin in submandibular gland cells of SS mice.
Abstract:ObjectiveTo illustrate the effect of M1/M2 polarization of macrophages on gouty arthritis models induced with monosodium urate and reveal the molecular mechanism of total saponins from Dioscoreae Nipponicae Rhizoma to treat gouty arthritis.MethodA total of 72 male SD rats were randomly divided into four groups: normal group, model group, total saponin group (160 mg·kg-1), celecoxib group (43.3 mg·kg-1), with 18 rats in each group. Gouty arthritis models were induced by injecting monosodium urate into ankle joints bilaterally. Histopathology changes of ankle joints were observed by hematoxylin-eosin(HE) staining. Immunohistochemistry method was used to detect the protein expression change of CD68, interleukin-4(IL-4), inducible nitric oxide synthase (iNOS) and transforming growth factor-β1(TGF-β1).ResultHE staining results showed that the inflammation of the model group was most obvious on the third day after modeling, and the disease was in the acute stage. On day 5, the inflammation was alleviated, and on day 8, the inflammation was still present but close to normal. The total saponin group and celecoxib group could improve the pathological changes of synovial tissue, and the effect of total saponin group was more obvious. Immunohistochemical results were as follows. Compared with the normal group. The expression of CD68 and iNOS in the model group increased on the 3rd,5th and 8th day of administration (P<0.01). Compared with the model group, the total saponins group could reduce the expression of CD68 and iNOS (P<0.05,P<0.01)on the 3rd day of administration, and significantly reduced them expression on the 5th and 8th days (P<0.01). Compared with the normal group, IL-4 and TGF-β1 expression were increased in the model group when the drug was given for three days(P<0.01). Total saponin group could enhance IL-4 expression(P<0.05)and decreased the TGF-β1 expression(P<0.01). Compared with normal group, the expression of IL-4 in the model group decreased on the 5th and 8th day of administration (P<0.01), and the expression of TGF-β1 in the model group decreased on the 5th day of administration(P<0.01). Compared with the model group, the total saponins group could increase the expression of IL-4 and TGF-β1 at 5 d and 8 d after administration (P<0.01).ConclusionTotal saponins from Dioscoreae Nipponicae Rhizoma has the potential effect to treat gouty arthritis by regulating M1/M2 polarization.
Abstract:ObjectiveTo explore the effect and underlying mechanism of koumine (Kou) at different concentrations (0, 100, 200, 400 μmol·L-1) on the proliferation and apoptosis of colorectal cancer HCT-116 cells.MethodAfter 24 hours of in vitro intervention with HCT-116 cells by Kou, cell counting kit-8 (CCK-8) assay was used to detect its effect on cell proliferation. Flow cytometry was used to detect cell cycle, apoptosis, and reactive oxygen species (ROS) expression. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of forkhead box O3a (FoxO3a). Cells were transfected with small interfering ribonucleic acid (siRNA). Western blot was employed to detect the protein expression of the FoxO3a target gene.ResultCompared with the conditions in the blank group, Kou treatment reduced the proliferation rate of HCT-116 cells (P<0.05, P<0.01) in a dose-dependent manner, caused cell cycle arrest in the G0/G1 phase, and induced the apoptosis of HCT-116 cells (P<0.05, P<0.01), which was positively correlated with the concentration of Kou. FoxO3a siRNA interference reduced the expression of FoxO3a and its downstream target genes cyclin-dependent kinase inhibitor 1A (p21), cyclin-dependent kinase inhibitor 1B (p27), and Bcl-2 interacting mediator of cell death (Bim) (P<0.01). Kou treatment induced the activation of c-Jun N-terminal kinase (JNK) in HCT116 cells. SP600125 (JNK specific inhibitor) treatment inhibited the Kou-induced FoxO3a activation and the expression of its downstream target genes. N-acetyl cysteine (NAC) treatment reduced Kou-induced ROS levels (P<0.01) and JNK signal activation. The above results were significantly different from those in the blank group (P<0.01).ConclusionKou can effectively inhibit the proliferation of HCT-116 cells and promote apoptosis, and the mechanism may be related to the regulation of the ROS/JNK/FoxO3a pathway.
Abstract:ObjectiveTo observe the effect of oxymatrine (OM) combined with bevacizumab ( BV ) on the proliferation, invasion, and migration of breast cancer MCF-7 cells and explore the mechanism of OM in regulating BV-induced epithelial-mesenchymal transition (EMT) based on the Wnt/β-catenin signaling pathway.MethodThe effect of different concentrations of OM(0, 0.5, 1.0, 2.0, 4.0, 8.0, 16.0 mmol·L-1)and BV(0, 0.25×10-4, 0.50×10-4, 1.00×10-4, 2.00×10-4, 4.00×10-4, and 8.00×10-4 mmol·L-1)on the proliferation of MCF-7 cells were detected by cell counting kit-8(CCK-8)assay. The effect of OM(4.0 mmol·L-1) combined with BV(2.00×10-4 mmol·L-1)on the invasion and migration of MCF-7 cells were observed in transwell and scratch repair tests. Western blot was conducted to investigate the effect of OM(4.0 mmol·L-1)combined with BV (2.00×10-4 mmol·L-1) on proliferation-related proteins in MCF-7 cells, followed by the detection of the expression levels of Wnt/β-catenin signaling pathway- and EMT-related proteins.ResultCompared with the blank group, OM (2.0,4.0,8.0,16.0 mmol·L-1) inhibited the proliferation of MCF-7 cells in a concentration-dependent manner (P<0.01), while BV did not show the inhibitory effect against the proliferation of MCF-7 cells. The inhibitory effect of the combination of the two drugs on the proliferation of MCF-7 cells was not significantly different from that of OM. Compared with the blank group, OM significantly reduced the migration distance of MCF-7 cells and the number of invaded cells(P<0.01), while BV increased the migration distance of MCF-7 cells and the number of invaded cells (P<0.05,P<0.01). Compared with BV, its combination with OM significantly inhibited the invasion and migration of MCF-7 cells induced by BV (P<0.01). Compared with the blank group, both OM and the combined medication obviously inhibited the phosphorylation of proliferation-related protein kinase B(Akt) and extracellular-signal-regulated protein kinase 1/2 (ERK1/2)in MCF-7 cells (P<0.01) and down-regulated the protein expression levels of β-catenin, proto-oncogene (c-Myc), CD44, and G1/S-specific cyclin D1 in Wnt/β-catenin signaling pathway (P<0.05,P<0.01). Besides, OM and the combination of two drugs both significantly reduced the protein expression levels of calcium-dependent cell adhesion protein N-cadherin and Vimentin in EMT, whereas increased the expression of calcium-dependent cell adhesion protein E-cadherin(P<0.01). However, the expression of the above-mentioned proteins in the BV group was reversed (P<0.05,P<0.01).ConclusionAfter the combination with BV, OM plays an anti-breast cancer role by effectively inhibiting the activation of Wnt/β-catenin pathway induced by BV and reversing EMT.
Abstract:ObjectiveTo evaluate the clinical efficacy of Bupi Qingfei decoction in the treatment of bronchiectasis colonized by Pseudomonas aeruginosa (PA) (lung-spleen Qi deficiency syndrome and phlegm heat accumulating in lung syndrome).MethodA total of 72 bronchiectasis patients colonized with PA ( lung-spleen Qi deficiency syndrome and phlegm heat accumulating in lung syndrome ) were randomly divided into the observation group (36 cases, two cases were lost to follow-up and three dropped out) and control group (36 cases, three cases were lost to follow-up and four dropped out). There were 31 cases in the observation group and 29 cases in the control group completing the trial. Patients in the observation group were treated with Bupi Qingfei decoction orally,once in the morning and again in the evening, one bag every other day, and simulated azithromycin tablet at the dose of 0.5 g,once every other day, while those in the control group with azithromycin tablet at 0.5 g,once every other day, and simulated Bupi Qingfei decoction, once in the morning and again in the evening, one bag every other day. Patients in both groups received health education and postural expectoration. The treatment lasted for 24 weeks,followed by a 24-week follow-up. The frequency of acute exacerbation,quality of life (St. George's Respiratory Questionnaire) score,traditional Chinese medicine (TCM) syndrome score,lung function [forced expiratory volume in one second percentage of predicted(FEV1%pred) and FEV1/forced vital capacity(FVC)], and serum immunoglobulin (Ig)A,IgE,IgG,and IgM levels of the two groups were evaluated after treatment.ResultThe frequencies of acute exacerbation after 24 weeks of treatment and during the 24-week follow-up in the observation group were lower than those in the control group (P<0.05). The total quality of life (St. George's Respiratory Questionnaire) score and symptom scores in the observation group after 24 weeks of treatment were significantly decreased as compared with those before treatment (P<0.05). There was no significant improvement in the quality of life in the control group either after 24 weeks of treatment or during the 24-week follow-up. The effective rate against TCM syndrome in the observation group was 64.52%(20/31) after 12 weeks of treatment,which was obviously higher than 31.03%(9/29) in the control group (χ2=6.726,P<0.05). After 24 weeks of treatment,the effective rate in the observation group was 83.87%, slightly higher than 68.97% in the control group. After 12 and 24 weeks of treatment,the scores of cough,expectoration,fatigue,anorexia,spontaneous sweating,abdominal distension, and loose stool in the observation group were better than those in the control group (P<0.05). There were no significant changes in lung function and serum immunoglobulin classes in the two groups.ConclusionBupi Qingfei decoction is effective in reducing the frequency of acute exacerbation, alleviating the symptoms, and improving the quality of life of bronchiectasis patients colonized by PA (lung-spleen Qi deficiency syndrome and phlegm heat accumulating in lung syndrome).
Keywords:bronchiectasis;Pseudomonas aeruginosa (PA);Bupi Qingfei decoction;azithromycin;frequency of acute exacerbation;quality of life
Abstract:ObjectiveTo observe the clinical effect of Tanreqing injection combined with western medicine on pneumonia caused by multi-drug resistant bacteria (MDRB) in elderly patients.MethodA total of 140 MDRB-induced pneumonia inpatients with the syndrome of phlegm-heat obstructing lung in the intensive care unit (ICU) of Dalian Hospital of Traditional Chinese Medicine from December 2018 to December 2020 were divided into an observation group (70 cases) and a control group (70 cases)) according to the random number table method. The patients in the control group received conventional treatment by western medicine, and those in the observation group received conventional treatment by western medicine combined with Tanreqing injection. The course of treatment was 7 days. The main efficacy indexes of the two groups before and after treatment were recorded,including the total clinical efficacy of traditional Chinese medicine (TCM) syndrome,total TCM syndrome score,clinical pulmonary infection score (CPIS), and the clearance rate of MDRB. Secondary efficacy indexes included temperature recovery and cough remission time,procalcitonin (PCT),C-reactive protein (CRP),white blood cell count (WBC),interleukin-6 (IL-6),interleukin-8 (IL-8), and oxygen partial pressure (PO2).ResultThe total effective rates of the observation group and the control group were 90.00% (63/70) and 75.70% (53/70),respectively,and the observation group had superior curative efficacy (Z=-2.147,P<0.05). After treatment,CPIS and total TCM syndrome scores in both groups decreased compared with those before treatment,and the decrease was more significant in the observation group (P<0.01). The clearance rate of MDRB in the observation group was 67.1% (47/70),superior to 48.6% (34/70) in the control group (χ2=4.951,P<0.05). The temperature recovery and cough remission time in the observation group was shorter than that in the control group (P<0.01). After treatment,the levels of PCT,CRP,WBC,IL-6, and IL-8 in both groups were reduced compared with those before treatment,while the levels of PO2 increased (P<0.01). The improvement of various inflammatory indexes and the PO2 level in the observation group was better than that in the control group (P<0.01).ConclusionThe clinical efficacy of Tanreqing injection combined with western medicine in the treatment of MDRB-induced pneumonia in elderly patients is significant,which can control infection,reduce inflammatory damage,improve the clearance rate of MDRB and PO2,and alleviate clinical symptoms. It is worthy of clinical application.
Abstract:ObjectiveAccording to the GB/T 15000.3-2008, to develop a fucosterol certified reference material based on the project approved by Standardization Administration.MethodFucosterol was isolated from Laminaria japonica dried thallus via 95% ethanol extraction, vacuum concentration, repeated column chromatography separation, recrystallization in petroleum ether-ethyl acetate, and residual solvent removal. Its chemical structure was identified by elemental analysis (EA), infrared spectrum (IR), mass spectrometry (MS), nuclear magnetic resonance (NMR) and X-ray diffraction (XRD). Its homogeneity, stability, and cooperative certification conducted by 8 laboratories were carried out by high performance liquid chromatography with evaporative light scattering detector.ResultFor the fucosterol reference material, the certified value of purity was 99.54% with expanded uncertainty of 0.16% in confidence interval of 95%, the stability was good within 24 months storage period at 2-4 ℃, which met the technical requirements of reference material and passed the acceptance organized by Standardization Administration.ConclusionThe national standard materials of fucosterol has been successfully developed, which can be used for the determination of this component, the evaluation of detection methods, and the detection and quality control of related products.
Abstract:ObjectiveTo explore the high-efficiency and high-quality seedling raising method of Codonopsis pilosula.MethodIn the main production area of C. pilosula in the Tanchang county,Gansu province,after the soil was fumigated with dazomet (containing 98% methylisothiocyanate), four varieties of C. pilosula seedlings were raised. The dynamic change in growth and differences in quality and yield of C. pilosula seedlings after emergence were regularly determined.Result① The soil enzyme activity was first inhibited and then restored by soil fumigation,which increased the root length of C. pilosula seedlings by 9.8%. Besides, the field growth indexes such as plant height,plant width,stem length,stem diameter,number of branches,number of nodes,number of leaves, and fitted leaf area increased in varying degrees,and the plant height showed an "S"-shaped growth trend. "Gandang No.1" and "Gandang No.2" grew better than "Weidang No.1" and "Tanchang control". ② Fumigation reduced the incidence rate of C. pilosula root in the field by 4.9%,and the incidence rates of "Gandang No.1" and "Gandang No.2" were significantly lower than those of "Weidang No.1" and "Tanchang control". ③ Fumigation increased the total number of C. pilosula seedlings by 6.15×105 plants·hm-2,of which the number of primary seedlings increased by 45.3% and that of secondary seedlings increased by 42.2%. ④ Fumigation increased the seedling yield of C. pilosula by 42.4%. It showed the most significant effect on the yield of "Gandang No.2",which increased by 61.8%, and the weakest effect on the yield of "Gandang No.1",which increased by 15.4%. ⑤ Comprehensive analysis showed that the quality and yield of C. pilosula seedlings in the fumigation area were better than those in the non-fumigation area.ConclusionThe results showed that soil fumigation showed a promoting effect on the seedling yield of C. pilosula in spite of different effects achieved in terms of different varieties.
Abstract:ObjectiveTo investigate the biological essence of the content variation of differential primary and secondary metabolites in fresh roots of Scutellaria baicalensis under drought stress.MethodThe changes of metabolites were analyzed by ultra-high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass/mass (UHPLC-ESI-Q-TOF-MS/MS).ResultA total of 11 differential compounds were identified from the roots of S. baicalensis (VIP≥2). Under drought stress, citric acid content increased and shikimic acid content decreased, indicating that the drought stress weakened the primary metabolism but strengthened secondary metabolism. Drought stress raised the content and regulated the proportion of various secondary metabolites by modulating the biosynthesis and biotransformation of them. To be specific, the content of free flavonoids with many phenolic hydroxyl groups and high biological activity and pharmacological activity, such as baicalin, wogonoside, baicalein, wogonin, chrysin, eriodictyol, 5,2',6'-trihydroxy-7,8-dimethoxyflavone, 5,8-dihydroxy-6,7-dimethoxyflavone, and 3,5,7,2',6'-pentahydroxyflavanone, was significantly increased. The massive compounds, like an intricate buffer, maintain metabolism stable as quickly and accurately as possible through biosynthesis and biotransformation, thus responding to the changing environment, which reveals how the quality of genuine regional drugs is influenced and why compounds in herbal medicine are complex.ConclusionSecondary metabolites with low content but high activity are important influencing factors of medicinal material quality and metabolites with high content and high activity are evaluation indicators of genuine regional drug quality.
Abstract:ObjectiveTo establish ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for simultaneous determination of six hepatotoxic pyrrolizidine alkaloids in Verbenae Herba, and to carry out preliminary risk assessment according to the research results.MethodAn ACQUITY UPLC HSS T3 column (2.1 mm×100 mm, 1.8 μm) was used for analysis with 0.05% formic acid and 2.5 mmol·L-1 ammonium formate in water (A)-0.05% formic acid and 2.5 mmol·L-1 ammonium formate in acetonitrile (B) as mobile phase for gradient elution (0-12 min, 3%-8%B; 12-25 min, 8%-15%B; 25-26 min, 15%-3%B; 26-30 min, 3%B), the flow rate was 0.3 mL·min-1, the column temperature was 40 ℃, and the injection volume was 1 μL. MS system was operated by electrospray ionization (ESI) in the positive ion mode with multiple reaction monitoring mode. MS parameters of triple quadrupole and six analytes were optimized for qualitative and quantitative analysis. According to the determination results, the risk assessment was carried out by using margin of exposure (MOE) combined with transfer rate of hot water extraction.ResultBased on the instrument precision, linear range, repeatability, stability, recovery and other methodological validations, the results were in conformity with relevant standards of quantitative analysis. The linear ranges of intermedine, lycopsamine, intermedine N-oxide, lycopsamine N-oxide, echimidine N-oxide and echimidine were good (r≥0.999 0) between peak area and mass concentration in the ranges of 0.984-49.20, 0.994-49.70, 1.012-50.60, 1.032-51.60, 1.004-50.20, 1.016-50.80 µg·L-1, respectively. The average recoveries of these six analytes were 87.2%-94.2% with relative standard deviation (RSD)<4.0%. Their MOE values were >10 000.ConclusionThe UPLC-MS/MS established in this study is stable and feasible, which can provide scientific basis for the quality control and safety evaluation of hepatotoxic pyrrolizidine alkaloids in Verbenae Herba.
Keywords:Verbenae Herba;hepatotoxic components;pyrrolizidine alkaloids (PAs);ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS);determination;risk assessment;margin of exposure (MOE)
Abstract:ObjectiveTo explore the effects of anti-microbial compound (T1) from Bacillus (Phylum Firmicutes) and anti-microbial compound (T2) from Pseudomonas and Rhizobium, two growth-promoting agents, on the physiological characteristics and growth of Fritillaria przewalskii, in order to lay a foundation for the development of functional microbial agents and the promotion of ecological planting.MethodThe endophytic bacteria of F. przewalskii were isolated and identified using conventional methods. The leaves of three-year-old F. przewalskii were sprayed with T1 and T2, followed by yield determination. The enzyme activities and physiological and biochemical indexes in the plant and microorganisms were measured using the corresponding assay kits, and the contents of related hormones by liquid chromatography-mass spectrometry (LC-MS).ResultThe isolated endophytic bacteria were classified into Firmicutes,Proteobacteria, and Actinomycetes. The activities of superoxide dismutase(SOD) and peroxidase(POD) and auxin content after T2 treatment were significantly higher than those after T1 treatment, while the contents of siderophore,salicylic acid, and gibberellin were lower. Compared with the blank (CK) group, T1 and T2 increased the contents of endogenous gibberellin,cytokinin, and auxin in F. przewalskii leaves,but did not significantly change jasmonic acid and abscisic acid. T1 promoted the accumulation of endogenous salicylic acid in F. przewalskii leaves, but there was no significant change after T2 treatment. Compared with CK,T1 and T2 enhanced the activities of SOD, POD, and catalase (CAT) and decreased the content of malondialdehyde. T2 promoted the accumulation of hydrogen peroxide in F. przewalskii leaves, but no significant difference was observed after T1 treatment. Compared with CK,both T1 and T2 increased chlorophyll,average iron content in rhizosphere soil, and 100-plant weight.ConclusionT1 and T2 treatments help to increase the yield,and their specific mechanisms differ from each other. T1 exhibits better effect than T2.
Abstract:ObjectiveAiming at the residue of Shaoyao Gancaotang, the extraction, qualitative and quantitative study of the small molecule resource components were carried out to clarify the residual small molecule chemical components in the residue and explore the ways of its resource utilization.MethodThe ultra-performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS) was used to qualitatively identify the residual small molecule substances in the dregs of Shaoyao Gancaotang. Agilent C18 reversed-phase chromatographic column (3.0 mm×100 mm, 2.7 µm) was used at the flow rate of 0.4 mL·min-1, the injection volume was 5 µL, and the mobile phase was gradient eluted with 0.05% formic acid aqueous solution (A)-acetonitrile (B) (0-1 min, 14%-17.5%B; 1-3 min, 17.5%-19%B; 3-4 min, 19%-20%B; 4-5 min, 20%B; 5-6 min, 20%-21%B; 6-9 min, 21%B; 9-22 min, 21%-36%B; 22-23 min, 36%B; 23-32 min, 36%-43%B), electrospray ionization (ESI) was employed with negative ion mode scanning and scanning range of m/z 50-1 200. A high performance liquid chromatography (HPLC) was established for the quantitative analysis of its main components with Agilent C18 reversed-phase chromatographic column (4.6 mm×150 mm, 5 µm), the detection wavelength was set at 235 nm, the flow rate was 0.8 mL·min-1, and the injection volume was 5 µL. Mobile phase was 0.05% phosphoric acid (A)-acetonitrile (B) for gradient elution (0-1 min, 14%-19%B; 1-4 min, 19%B; 4-18 min, 19%-50%B). The content changes of main components in the residue of Shaoyao Gancaotang were compared before and after two different techniques of organic solvent extraction and enzymatic extraction.ResultA total of 16 chemical components in the residue of Shaoyao Gancaotang were qualitatively analyzed, and quantitative analysis found that there were many chemical components in the residue, among which the residues of 6 index components such as paeoniflorin and liquiritin reached more than 70% in the original decoction piece. After enzymolysis by cellulase, liquiritin in the residue could be converted into liquiritigenin. The content of crude polysaccharide in enzymatic extract of the residue was 6 times higher than that in the blank group, and the content was up to 12%.ConclusionThere are still many small molecule resource components in the residue of Shaoyao Gancaotang, which has great development potential. Organic solvents can be used to re-extract the target components in the residue, and liquiritin can be converted into liquiritigenin by biological fermentation technology, and the crude polysaccharide from the residue can be extracted by enzymatic method to develop animal feed. This study can provide reference basis and approach for reusing the residues of Shaoyao Gancaotang preparations and dispensing granules, so as to realize the high-value utilization of Shaoyao Gancaotang.
Keywords:Shaoyao Gancaotang;residue;resource components;ultra-performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry (UPLC-QTOF-MS/MS);crude polysaccharide;high performance liquid chromatography (HPLC);enzymatic hydrolysis
Abstract:ObjectiveTo explore the potential targets and pathways of steroid alkaloids from Solanum nigrum (SASN) in the treatment of non-small cell lung cancer (NSCLC) and analyze the possible mechanism.MethodThe active SASN against NSCLC were searched from literature. Then potential targets of SASN were screened through SwissTargetPrediction and PharmMapper, and those of NSCLC through GeneCards. Venny was employed to yield the common targets of the two, and Cytoscape to construct the 'medicinal-component-disease-target' network. Metascape was applied to enrich the Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways of the common targets, and STRING was used to generate the protein-protein interaction (PPI) network, followed by screening of key targets by Cytoscape. Finally, Western blot was used to verify the effects of the medicinal on key targets.ResultA total of 6 active SASN were screened out: solasonine, solamargine, solasodine, solanocapsine, solanidine, and N-methylsolasodine, which had 96 potential anti-NSCLC targets. These targets mainly involved the pathways in cancer, proteoglycans in cancer, and Forkhead box protein O (FoxO) pathway. PPI network analysis demonstrated 15 key anti-NSCLC targets of SASN, such as mitogen-activated protein kinase (MAPK)1, MAPK8, MAPK14, protein kinase B (Akt1), signal transducer and activator of transcription 3 (STAT3), and proto-oncogene tyrosine protein kinase (SRC). Meanwhile, Western blot results showed that SASN could significantly down-regulate the expression of the key proteins Akt1, SRC, and STAT3.ConclusionWe predicted the potential targets and pathways of SASN against NSCLC and obtained 15 key targets, from which we selected three key proteins for validation. The validation results were consistent with the prediction results. This paper is expected to lay a scientific basis for the subsequent in-depth study of the mechanisms of SASN against NSCLC.
Abstract:ObjectiveTo explore the mechanism and compatibility characteristics of Baimai ointment (BMO) in the treatment of white vein disease from the network perspective based on system theory, so as to provide biological basis for its clinical application.MethodThe chemical components and the corresponding candidate target spectra of BMO were obtained from The Encyclopedia of Traditional Chinese Medicine (ETCM) and Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine (TCMIP). According to the clinicopathological characteristics of white vein disease, focusing on four diseases/symptoms including neuropathic pain, inflammatory pain, chronic pain and lumbar disc herniation root neuralgia, the gene sets related to white vein disease were collected in Human Phenotype Ontology (HPO), DisGeNET and other databases, then the interaction network of the targets of active components in BMO-gene sets related to white vein disease was constructed. On this basis, the hub network nodes were selected and enriched for exploring the mechanism of four functional groups of BMO in the treatment of white vein disease such as Huoxue Tongluo group (Curcumae Longae Rhizoma, Moschus, Tronae), Xingqi Zhitong group (Myristicae Semen, Nardostachyos Radix et Rhizoma, Acori Calami Rhizoma), Wenjing Sanhan Tongluo group (Zingiberis Rhizoma, Zanthoxyli Pericarpium, Caraway) and Jianpi Wenshen Qianggu group (Actinolite, Glycyrrhizae Radix et Rhizoma).ResultThe enriched pathways of the four functional groups in BMO were mainly distributed in three modules of nervous system function, inflammation-immune system regulation and body energy metabolism, and each module was connected by common target genes especially had its own focus. Among them, the regulation of nervous system function in Huoxue Tongluo group and Xingqi Zhitong group could be summarized as Huoxue Buqi and Xingshen Kaiqiao. Xingqi Zhitong group and Jianpi Wenshen Qianggu group were mainly used to promote the operation of Qi, promote blood metaplasia, enhance immunity and maintain the regulation of inflammation-immune system. Jianpi Wenshen Qianggu group and Wenjing Sanhan Tongluo group mainly regulated body energy metabolism by invigorating the spleen and supplementing Qi as well as warm-heat medicine. The whole formula focused on the multi-dimensional and multi-level mechanism of BMO in the intervention of white vein disease. Each functional group emphasized its respective characteristics in nervous system function, inflammation-immune regulation, and body energy metabolism. Two types of networks analysis models complemented and verified each other.ConclusionBMO plays a role in the treatment of white vein disease mainly by regulating the function of nervous system, maintaining the balance of inflammation-immune system and interfering with energy metabolism. The relevant research results can provide reference for the in-depth exploration of the mechanism of BMO, and help to guide the clinical rational use of this preparation.
Abstract:ObjectiveTo explore the effective components, targets, and possible mechanisms of Wenshen Yangxue prescription in improving endometrial receptivity of aged female mice based on network pharmacology and experimental verification.MethodBased on Bioinformatics Analysis Tool for Molecular mechANism of Traditional Chinese Medicine (BATMAN-TCM) and Integrative Pharmacology-based Research Platform of Traditional Chinese Medicine, the components and targets of Wenshen Yangxue prescription were retrieved, and the targets of ovulatory dysfunctional infertility were collected from the Online Mendelian Inheritance in Man (OMIM) and GeneCards with "anovulatory sterility" and "anovulatory infertility" as keywords. The protein-protein interaction (PPI) network was constructed based on STRING and the core targets of Wenshen Yangxue prescription against ovulatory dysfunctional infertility were screened by Cytoscape, followed by Gene Ontology (GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment of the core targets in DAVID database. Then, the "medicinal-component-target-pathway" network was established and the core targets were verified by animal experiment.ResultA total of 253 components and 326 targets of Wenshen Yangxue prescription, 819 disease targets, and 74 common targets were screened out. The common targets were mainly involved in the biological processes such as positive regulation of nitric oxide biosynthetic process, positive regulation of cell proliferation, response to estradiol, aging, response to oxidative stress, and angiogenesis. The GO term of response to oxidative stress and five of the top 20 KEGG pathways were analyzed. According to the "medicinal-component-target-biological process/pathway" network, 41 chemical components in 20 medicinals participated in hypoxia inducible factor-1 (HIF-1) signaling pathway, tumor necrosis factor (TNF) signaling pathway, forkhead box O (FOXO) signaling pathway, Toll-like receptor (TLR) signal pathway, and phosphatidylinositol 3-kinase/protein kinase B (PI3K/Akt) signaling pathway by affecting 35 targets. The results of animal experiment showed that the prescription could increase the expression of PI3K, phosphorylated PI3K (p-PI3K), Akt, phosphorylated Akt (p-Akt), forkhead box O3A (FoxO3A), and phosphorylated FoxO3A (p-FoxO3A) in uterus of aged female ICR mice.ConclusionWenshen Yangxue prescription interferes with oxidative stress and PI3K/Akt/FoxO3A signaling pathway by influencing Akt1, dual oxidase 2 (DUOX2), epidermal growth factor receptor (EGFR), heme oxygenase-1 (HMOX1), myeloperoxidase (MPO), and other targets, thereby improving endometrial receptivity of aged female mice.
Abstract:Literature research on ancient books of traditional Chinese medicine (TCM) is an important carrier for inheriting the academic achievements and thoughts of TCM, and a key step for continuing the Chinese civilization and realizing the great rejuvenation of the Chinese nation. Based on this, the paper puts forward the purpose of sorting out TCM ancient books:to explore the treasure of traditional culture, reveal its significance, carry forward its spirit, learn from its experiences, so as to make a contribution to the development of TCM. And this paper expounds several major problems in the literature research, that is, paying attention to the phenomenon of "stubborn bass", avoiding the trend of "latecomers turning inferior", attaching importance to the hidden trouble of "making comments on behalf of the ancients". Then, this paper discusses the methodology of carrying out accurate research and revealing the true nature and true value of scholarship with the idea of confucian orthodoxy, the rules of not forgetting the original intention and the scientific method. Taking the materia medica archaeology as an example, the author shared the practical exploration of how to crack the historical code with scientific and technological means, so as to provide useful reference for the literature research on TCM ancient books.
Keywords:traditional Chinese medicine;ancient books;collation;literature;textual research
Keywords:diabetic macroangiopathy;intestinal microecology;host immunity;inflammation;disorders of glucose and lipid metabolism;Superabundance of Yangming
Abstract:As common and frequently-occurring disorders in clinic practice,renal diseases are characterized by the impairment of kidney structure and function due to a variety of reasons and can be divided into primary,secondary, and hereditary types. Clinically,the impairment of kidney structure and function is usually a chronic progressive process,and the resulting chronic renal diseases have become a major public health problem endangering human health worldwide. Notch signaling pathway affects cell proliferation,differentiation,migration,growth, and apoptosis and determines the fate of cells. Abnormal expression or gene mutation of Notch will cause tissue damage, followed by the occurrence and development of a variety of renal diseases. Traditional Chinese medicine (TCM), as an important means to prevent and treat renal diseases,has the characteristics of acting on multiple targets and signaling pathways with multiple components,and is often used as a routine or potential complementary therapy for the treatment of chronic renal diseases and also a source of new drug discovery. In recent years, considering the limitations of western medicine in treating renal diseases,more and more scholars have begun to take Notch signaling pathway as the breakthrough point for exploring TCM prevention and treatment of renal diseases. They have conducted clinical and experimental studies on the regulation of Notch signaling pathway by a variety of individual Chinese herbs or their extracts,Chinese patent medicines, and Chinese medicinal compounds,and found that TCM exerted the renal protective effects by inhibiting the Notch signaling pathway. By collecting relevant literatures on TCM prevention and treatment of various renal diseases,especially those concerning TCM regulation of Notch signaling pathway for preventing and treating such chronic renal diseases as diabetic nephropathy,immunoglobulin A (IgA) nephropathy,renal fibrosis,membranous nephropathy,focal segmental glomerulosclerosis, and renal cell carcinoma,this paper summarized the current research status,in order to provide reference for clinical prevention and treatment of various renal diseases and build up the factual basis for the universal application of TCM.
Keywords:renal diseases;Notch;traditional Chinese medicine (TCM);diabetic nephropathy;immunoglobulin A (IgA) nephropathy;renal fibrosis;review
Abstract:Cerebral ischemia/reperfusion injury (CIRI) is a common feature and the main pathophysiological mechanism of ischemic stroke(IS), which is caused by a blood reperfusion injury in ischemic brain tissues. It can aggravate brain tissue injury and cause irreversible brain damage, seriously affecting the quality of life or even the life of patients. Hence, we must find out the exact mechanism as well as the effective therapeutic drugs and targets for CIRI. The Chinese medicine effective in Xingnao (restoring consciousness) and Kaiqiao (opening orifices) has been widely used in the treatment of CIRI and serves as a classic therapy for IS. In recent years, scholars have conducted extensive and in-depth studies on the mechanism and therapeutic targets of Chinese medicine in Xingnao and Kaiqiao. They found that those drugs could interfere with a series of changes after IS and achieve the remarkable curative effect. This study summarized the effect and mechanism of Chinese medicine in Xingnao and Kaiqiao in the treatment of CIRI, including reducing the inflammatory response and oxidative stress, alleviating brain edema and the toxicity of excitatory amino acids, reducing cell apoptosis, promoting angiogenesis and neurovascular remodeling, and improving blood-brain barrier injury. It is expected to provide references to clarify the mechanism and important targets of those drugs in resisting CIRI and ideas for the in-depth investigation and application of brain protection of Chinese medicine in Xingnao and Kaiqiao.
Keywords:Chinese medicine in Xingnao and Kaiqiao;cerebral ischemia/reperfusion injury(CIRI);neuroprotective effect;mechanism of action
Abstract:According to the 2020 world cancer data report, the prevalence of prostate cancer ranks second in male malignant tumors and the mortality fifth. In China, due to the poor living and eating habits, the prevalence of prostate cancer is still rising, and advanced prostate cancer can adapt to and tolerate androgen castration related drugs through a variety of mechanisms, which often indicates poor therapeutic outcomes in the late stage. Houttuyniae Herba, a medicinal plant of family Saururaceae, has been officially identified both as food and medicine by the National Health Commission. The existing studies have shown that Houttuyniae Herba inhibits the proliferation, migration, and invasion of prostate cancer and induces its apoptosis and cell cycle arrest via multiple targets, links, and pathways. These anti-tumor activities can also be observed in breast cancer, leukemia, gastric cancer, colorectal cancer, and other tumor cells. Such activities are mainly related to the inhibited expression of cyclin and cyclin-dependent kinase, regulation of apoptosis-related proteins, activation of apoptosis-related pathways, reduced secretion of matrix metalloproteinase, inhibition of activator of transcription 3 (STAT3)/Zinc finger protein (Snail) / transcription factor twist signaling pathway, and antagonization of growth factor receptor, androgen, inflammation, and oxidative stress in promoting the occurrence and development of prostate cancer. In addition, Houttuyniae Herba is able to alleviate diabetes, regulate blood glucose-lowering pathways, and boost immunity by inhibiting the protein kinase B (Akt)/mammalian target of rapamycin (mTOR) and mitogen-activated protein kinase (MAPK) signaling pathways, thus suppressing the occurrence and progression of prostatic cancer. In conclusion,Houttuyniae Herba can be used as a potential drug against prostate cancer.