Abstract:Quality marker(Q-marker) is a new concept and pattern for quality control of traditional Chinese medicine(TCM), which will lead the development direction for quality control of TCM. Among them, how to characterize the overall quality attribute of TCM and its biological effect, is a critical scientific problem in the study of Q-marker. In this paper, integrated pharmacology is utilized to screen out and confirm the Q-marker from the complex system of TCM, so as to solve the critical scientific problem. System biology in vivo is firstly applied to establish the correlation of chemical fingerprints of TCM, their metabolic fingerprints, network targets, biological effects and efficacy of TCM, which is used to preliminary screen out Q-marker of TCM. Following that, a pharmacological method in vitro, including intestinal absorption in vitro coupled with bioactivity assessment, is employed to simultaneously determine the absorbed doses of TCM and evaluate their biological activity. Furthermore, data mining is utilized to establish the exact quantitative mathematic model between Q-marker of TCM and bioactivity. Meanwhile, two representative examples, including Yuanhu Zhitong tablets, Xinsuning capsules, are introduced to identify Q-marker of TCM and establish their quality standards related with bioactivity, which will be beneficial to improve the level of quality control of TCM and ensure the effectiveness and safety of clinical applications.
Keywords:integrated pharmacology;quality markers of traditional Chinese medicine;in vitro pharmacology;exact quantitative association;component knockout technology;biological effect;Yuanhu Zhitong tablets
Abstract:The decreased efficacy and severe side effects of antibiotics, as well the increase of multidrug resistant pathogens are seriously threatening human health. It has become an urgent task for the whole world to actively respond to threats and establish effective prevention and control plans. Traditional Chinese medicine (TCM) has a long history with exact curative effect in the treatment of infectious diseases. It not only inhibits pathogens and eliminates endotoxin, but also has therapeutic effect on inflammation, immune abnormality and overall disorder caused by infection. Antibiotics mainly inhibit the pathogen itself, while the combination of TCM and antibiotics is complementary with each other and is considered as a feasible solution to the challenges. Some clinical observations have shown that TCM has potentiality of enhancing antibiotics efficiency and reducing toxicity. Exploring its mechanism is the necessary measure to optimize and popularize treatment regimen. Firstly, multi-level and multi-dimensional systematic pharmacology network analysis methods are used to predict the mechanism of TCM combined with antibiotics in the treatment of infectious diseases, so as to provide the evidence for further empirical research and selection of test indicators. Then by following the principle of corresponding drug use in specific TCM syndromes, the antibiotics and TCM are individually and jointly applied to treat patients with specific syndrome conditions of infectious diseases. Besides routine and comprehensive evaluation of synergistic and attenuated effect of the combined drug use, multi-omics technique is also used to find the subtle effects of these two drugs at a molecular level. The sensitive and stable clinical biomarkers of synergism and attenuation of combined drug use are determined by using biomolecular network analysis technology. Finally, taking these biomarkers as clues, the biotransformation process and regulation mechanism of the biomarkers are traced back in animal models of infectious diseases and cell level, and all of these are clinically verified. As a result, the mechanism of efficacy enhancing and toxicity reducing of combined drug use can be revealed, providing basis for the promotion and application of such combined drug use.
Keywords:traditional Chinese medicine;antibiotics;efficacy enhancing and toxicity reducing of the combined medication;mechanism study
Abstract:Objective:To observe the effects of recipes for tonifying kidney and replenishing Qi, Zuoguiwan (ZG) and Yiqi Congming Tang(YQ) on memory capacity, expressions of learning and memory-related genes expression, and explore the changes in relevant epigenetic modification enzymes.Method:SD male rats with natural aging (24 months old) were used as animal models and randomly divided into aged control group, aged ZG group(12.12 g·kg-1), aged YQ group(10.18 g·kg-1), aged compound group(11.15 g·kg-1) and aged antagonist RU38486 group(5×10-3g·kg-1). Another 5 months old male SD rats were included as the young control group. Morris water maze method was used to observe the spatial learning and memory ability of the rats. The co-localizations of histone deacetylase 2 (HDAC2) and methyl CpG binding protein 2 (MeCP2) in hippocampus of rats in each group were observed by laser confocal microscope. The changes in expressions of glucocorticoid receptor (GR), synapsin1(Syn-1), HDAC2, and histone acetyltransferase 1(HAT1) proteins in hippocampus of each group were detected by Western blot, and mRNA expression of HDAC2 was detected by Real-time fluorescence quantitative polymerase Chain reaction (Real-time PCR). At the same time, the effects of ZG, YQ and compound decoction in alleviating the above-mentioned abnormal changes were observed.Result:Compared with the young control group(control group), the latency of the aged control group was significantly prolonged (P<0.05), the number of crossing platforms was significantly reduced (P<0.05); compared with the aged control group(model group), the latencies of the ZG group and the YQ group were significantly shortened (P<0.05), and the numbers of crossing platforms were increased. Compared with the young control group, the protein and mRNA expressions of HDAC2 in the aged control group were significantly increased (P<0.05, P<0.01), and the co-expression with MeCP2 in the nucleus was significantly increased; and the expressions of HAT1, GR and Syn-1 were significantly decreased(P<0.05, P<0.01). ZG, YQ and compound can all alleviate the abnormal changes of the above indicators to varying degrees, but the YQ and the compound group had no effect on HAT1.Conclusion:ZG group, YQ group, and compound group can improve the spatial learning and memory abilities of aged rats by increasing the expression of learning-memory-associated protein GR and epigenetic modification enzyme HAT1, and reducing the expression of HDAC2 and the co-localization of HDAC2 protein and MeCP2 in the nucleus.
Keywords:aging;tonifying kidney decoction;Zuoguiwan;Yiqi decoction;Yiqi Congming Tang;learning and memory;epigenetic modification
Abstract:Objective:To observe the expression of mammalian target of rapamycin (mTOR) that mediates autophagy in pulmonary fibrosis and the effect of autophagy in the formation of pulmonary fibrosis, in order to explore the treatment mechanism of Buyang Huanwu Tang on pulmonary fibrosis.Method:Totally 144 C57BL/6 mice were randomly divided into 6 groups: sham operation group, model group, prednisone group, high-dose Buyang Huanwu Tang group, medium-dose Buyang Huanwu Tang group and low-dose Buyang Huanwu Tang group, with 24 mice in each group. The sham operation group was injected with the same amount of 0.9%saline. The remaining groups were treated with bleomycin tracheal injection to replicate the pulmonary fibrosis model. After modeling, sham operation group and model group were given 0.9%normal saline (0.01 g·kg-1·d-1), group high-dose Buyang Huanwu Tang group was given Buyang Huanwu Tang (28.08 g·kg-1·d-1), medium-dose Buyang Huanwu Tang group was given Buyang Huanwu Tang (14.04 g·kg-1·d-1), low-dose Buyang Huanwu Tang group was given Buyang Huanwu Tang(7.02 g·kg-1·d-1), and P group was given prednisone (0.455 g·kg-1·d-1) by gavage. The samples were taken in batches on the 7th, 14th and 28th days after modeling; degrees of alveolitis and fibrosis in mice were observed by hematoxylin-eosin (HE) staining and Masson staining. The mTOR protein, ribosomal S6 protein and microtubule associate protein 1 hight chain3-Ⅱ(MAP1LC3-Ⅱ) of mouse lung tissue were detected by Western blot; electron microscopy was used to observe the autophagy of lung tissue in mice.Result:Compared with the sham-operated group, the degrees of alveolitis and pulmonary fibrosis were significantly severer in the model group on 7th, 14th and 28th days (P<0.01); compared with the model group, the degrees of alveolitis and pulmonary fibrosis were alleviated at each observation time in the drug-administered groups (P<0.05). The decreased scores in high-dose Buyang Huanwu Tang group were the most obvious, with statistically significant differences (P<0.05). compared with the sham-operated group, the expressions of mTOR and S6 protein in lung tissue of the model group were significantly up-regulated, and each drug-administered group showed down-regulations. Compared with the sham-operated group, the expression of LC3-Ⅱ protein in lung tissue of the model group was significantly down-regulated, and each drug-administered group showed anup-regulation(P<0.01). The cells in the sham-operated group were well-formed, and no autophagy was observed. The model group had the worst cell morphology, with individual autophagy. The cell morphology and autophagy in each drug-administered group were better than those in the model group. The high-dose Buyang Huanwu Tang group and the medium-dose Buyang Huanwu Tang group had the highest number of autophagosome.Conclusion:The mTOR protein is activated in mice lung tissue, autophagy is inhibited, mTOR protein participates in the pathogenesis of pulmonary fibrosis by inhibiting autophagy; Buyang Huanwu Tang has a certain therapeutic effect on BLM-induced pulmonary fibrosis in mice, and its mechanism may be related to the down-regulation of mTOR protein expression that mediates autophagy.
Abstract:Objective:To explore the mechanism of modified Tianwang Buxindan on oxidative damage of Trx system in parachlorophenylalanine (PCPA) insomnia model rats.Method:Sixty male SD rats were randomly divided into blank group and model group. Insomnia model was prepared through intraperitoneal injection with PCPA (150 mg·kg-1). The discontinuous injection lasted for 7 d. After successful modeling, the rats were divided into model group (the same volume of normal saline), low, medium, high-dose Tianwang Buxindan groups (8.8, 17.6, 35.2 g·kg-1·d-1) and estazolam group (0.1mg·kg-1·d-1), with 10 in each group. Autonomous activity video was used to detect circadian activity rhythm. Transmission electron microscopy (TEM) was used to observe supra chiasmatic nucleus (SCN) morphology and Organelle integrity. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expressions of superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione peroxidase (GSH-Px) in serum. Expressions of Trx2, TrxR2 in SCN cells were detected by immunofluorescence (IF) and Western blot.Result:Compared with blank group, the activity rhythm of model group was irregular, the activity time increased (P<0.01), the mitochondrial cristae were broken, the number was reduced, the arrangement was disorder, SOD and GSH-PX decreased, MDA increased (P<0.01), the expression of Trx2 decreased, while the expression of TrxR2 increased (P<0.01). Compared with model group, medium and high-dose Tianwang Buxindan groups could alleviate the circadian rhythm disorder and reduce the activity time (P<0.01), the mitochondrial edema was relieved, part of the cristae were intact, the activities of SOD and GSH-PX were increased, the level of MDA was reduced (P<0.05, P<0.01), expression of Trx2 was significantly increased, and expression of TrxR2 was significantly reduced (P<0.05, P<0.01).Conclusion:The effect of modified Tianwang Buxindan on insomnia model rats is related to the regulation of Trx2 and TrxR2 protein expressions in Trx system.
Keywords:insomnia;Tianwang Buxindan;supra chiasmatic nucleus (SCN);oxidative stress;thioredoxin system
Abstract:Objective:To observe the effect of Wendantang on tumor necrosis factor-α (TNF-α), interleukin(IL)-6, IL-17, IL-22 and other related inflammatory factors in peripheral blood and the expression of STAT3[the key molecule of Janus kinase 2/signal transducer and activator of transcription 3(JAK2/STAT3) signal pathway in hypothalamus] mRNA and protein of obese rats with syndrome of phlegm-dampness, so as to explore the internal mechanism of Wendantang in interfering obesity with syndrome of phlegm-dampness.Method:A total of 100 rats were randomly divided into blank group(30 rats) and modeling group(70 rats), rats in the blank group was fed with basic feed and the modeling group was fed with high-fat feed for 6 weeks. Animal model of obesity with syndrome of phlegm-dampness was established by the method in literature. After successful modeling, 16 obese rats were selected and randomly divided into the model group and Wendantang intervention group with 8 rats in each group, and another 8 rats in the blank group were randomly selected as the normal group. Rats in Wendantang intervention group were given 15 g·kg-1 of crude drug by gavage, while the model group and the normal group were given the same amount of distilled water for gavage once a day for 6 weeks. No eating but no prohibiting drinking before dealing with 12 h and then taking samples after anesthesia. The body weight, Lee's index and obesity rate of rats were measured, the levels of blood lipids[total cholesterol(TC), triglyceride(TG), low density lipoprotein cholesterol(LDL-C) and high density lipoprotein cholesterol(HDL-C)] of rats were detected with a full-automatic biochemical analyzer according to the requirements of the kit, the expression of TNF-α, IL-17, IL-22 and IL-6 in peripheral blood of rats was detected with enzyme-linked immunosorbent assay(ELISA), STAT3 mRNA expression in hypothalamus of rats was detected with real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) and the expression of STAT3 protein in hypothalamus of rats was detected with Western blot. Result:The high-fat feed feeding could successfully replicate the obese rat model, and the obesity rate of rats in the modeling group was greater than 20%, and compared with the blank group, the body weight and Lee's index of rats in the modeling group were significantly increased(P<0.05, P<0.01). Compared with the normal group, the body weight of rats in the model group was significantly increased(P<0.01), blood lipid level changed greatly(P<0.01), levels of related inflammatory factors were significantly increased(P<0.01), the STAT3 mRNA and protein expression in hypothalamus tissue of rats was significantly increased(P<0.01). Compared with the model group, the body weight and Lee's index of rats in Wendantang intervention group significantly decreased(P<0.05, P<0.01), blood lipid level significantly changed(P<0.01), levels of related inflammatory factors were significantly decreased(P<0.01), the STAT3 mRNA and protein expression in hypothalamus tissue of rats in Wendantang intervention group was significantly decreased(P<0.01).Conclusion:Wendantang has a good effect on improving phlegm-dampness in obese rats, and the mechanism may be related to regulating JAK2/STAT3 signal pathway then to improve the chronic inflammatory state of the body, and all of which provides a scientific basis for Wendantang in intervening obesity with syndrome of phlegm-dampness.
Keywords:obesity with syndrome of phlegm-dampness;Wendantang;inflammatory factors;signal transducer and activator of transcription 3(STAT3);Janus kinase 2/STAT3(JAK2/STAT3) pathway;Lee's index;chronic inflammation
Abstract:Objective:To investigate the effect of copper ion(Cu2+ ) on the aggregation and neurotoxicity of Aβ, and affirm the role of Danggui Shaoyaosan in vitro, the Neuroblastoma (SH-SY5Y) cells treated with β-amyloid 1-42 (Aβ1-42) and Cu2+ were used as a vitro models of Alzheimer's disease(AD).Method:Aβ 1-42 (20 μmol·L-1) was reacted with different concentrations of copper sulfate (CuSO4, 20, 40 μmol·L-1), and then the thioflavine T (ThT) staining method was used to detect the Aβ aggregation state. The Aβ aggregation status was also detected by ThT staining in the Aβ1-42-Cu2+ group(20+ 20 μmol·L-1), and Danggui Shaoyaosan groups(1.6, 3.2, 6.4 mg·L-1). The SH-SY5Y cells were cultured and incubated with different concentrations of Aβ1-42(1.25, 2.5, 5, 10, 20, 40 μmol·L-1) and Danggui Shaoyaosan(1.6, 3.2, 6.4, 12.8 mg·L-1) for 24 h. Subsequently, SH-SY5Y cells were incubated with Aβ1-42 (20 μmol·L-1) and CuSO4(20 μmol·L-1) in the Aβ1-42-Cu2+ group, and incubated with Aβ1-42 (20 μmol·L-1), Danggui Shaoyaosan (1.6 mg·L-1) and CuSO4 (20 μmol·L-1) in Danggui Shaoyaosan group. Control group was added with the medium. After 24 h of co-action, the cell viability was detected by the methylthiazolyl tetrazolium (MTT) assay. The morphology of the cells was photographed by microscopy. The intracellular extracellular Aβ1-42 aggregation was detected by Western blot.Result:Cu2+ and Aβ1-42 bound to more and larger Aβ aggregates compared with the Aβ1-42 group. Compared with the normal group, cell viability was significantly reduced (P<0.05, P<0.01) and intracellular and extracellular Aβ1-42 aggregation was increased(P<0.05, P<0.01) in the model group. Danggui Shaoyaosan can increase the cell viability of SH-SY5Y damaged by Aβ-Cu2+ (P<0.05, P<0.01), and reduce Aβ1-42 protein (P<0.05, P<0.01).Conclusion:Cu2+ can increase the aggregation and toxicity of Aβ; Danggui Shaoyaosan can significantly reduce the damage of SH-SY5Y cells induced by Cu2+ -mediated Aβ aggregation, promote Aβ endocytosis, reduce extracellular Aβ aggregation and increase cell viability.
Abstract:Objective:To explore the hepatotoxic material basis of Polygoni Multiflori Radix with zebrafish model, in order to provide a theoretical basis for the study on the mechanism of Polygoni Multiflori Radix hepatotoxicity.Method:Emodin, rhein, aloe emodin, emodin-1-O-glucoside, physcion-8-O-glucoside and aloe emodin-8-O-glucoside for three days (at the concentrations of 0.000 73, 0.002 22, 0.015 05, 0.002 36, 0.198 95, 0.072 73 g·L-1) selected from the early stage of the experiment were continuously administered to zebrafish fertilized for 72 hours to establish the liver fluorescence transgenic larvae animal model. The activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), glutathione (GSH) and total bilirubin (TBIL) in zebrafish at 1, 2, 3 day after administration were measured respectively, and the pathological changes of zebrafish liver tissue were analyzed.Result:Emodin, rhein, emodin-1-O-glucoside and physcion-8-O-glucoside had no significant effect on the activities of ALT, AST, GSH and content of TBIL (P<0.01) in zebrafish, and the liver tissue of zebrafish was normal; aloe-emodin could significantly reduce the activities of ALT, AST, GSH (P<0.01), whereas increase the content of TBIL in zebrafish; aloe-emodin-8-O-glucoside could significantly reduce the activities of ALT, GSH, whereas increased the content of TBIL (P<0.05, P<0.01) in zebrafish, with no significant effect on the activity of AST, then both groups showed large flaky necrosis in zebrafish liver tissue, obvious morphologic changes in liver tissue cells, and irregular arrangement of hepatocytes.Conclusion:Aloe-emodin and aloe-emodin-8-O-glucoside in Polygoni Multiflori Radix have a toxic effect on zebrafish liver, suggesting that aloe-emodin and aloe-emodin-8-O-glucoside might be the hepatotoxic material basis of Polygoni Multiflori Radix.
Abstract:Objective:To investigate the effect and mechanism of Sanhuang Yinchi decoction (SHYCD) in preventing carbon tetrachloride (CCl4)-induced acute liver injury by regulating high mobility group box1(HMGB1) signaling pathway.Method:A total of 48 KM mice were randomly divided into blank control group, model group, low, middle and high-dose SHYCD groups and positive control group. The model of acute liver injury induced by CCl4 in mice was established. The low, middle and high-dose SHYCD groups were intragastrically administered with drugs (16, 32, 48 g·kg-1·d-1) respectively, and the positive control group was given cell growth stimulating hormone (20 mg·kg-1·d-1) through intraperitoneal injection. Pathological changes of mouse liver tissue sections were observed by hematoxylin-eosin staining (HE); relevant enzyme kits were used to determine liver function indexes in mice serum-alanine aminotransferase (AST) and aspartate aminotransferase (ALT); the expression level of interleukin-6 (IL-6) in mouse serum was determined by enzyme-linked immunosorbent assay (ELISA); Western blot was used to detect the expressions of high mobility group box-1(HMGB1), cysteine aspartic acid protease(Caspase-3), apoptosis-related molecules B cell lymphoma(Bcl-2), Bcl-2 associated x protein(Bax), and Toll-like receptor 4 (TLR4).Result:Compared with the normal group, the model group significantly increased serum AST, ALT (P<0.05) and IL-6 levels (P<0.05) and expressions of HMGB1, TLR4 and Caspase-3 (P<0.05), and down-regulated Bcl-2/Bax ratio (P<0.05) in liver tissue; compared with the model group, SHYCD can effectively alleviate the pathological damage of liver in mice, reduce serum AST and ALT levels and expressions of IL-6, HMGB1, TLR4 and Caspase-3 protein in liver homogenate (P<0.05), and increased the ratio of Bcl-2/Bax (P<0.05) in a dose-dependent manner.Conclusion:SHYCD can prevent liver injury by regulating HMGB1/TLR4/NF-κB signaling pathway, reducing cellular inflammatory response and inhibiting apoptosis, so as to prevent acute liver injury in mice. This indicates that HMGB1 may become a new target to prevent acute liver injury.
Keywords:acute liver injury;Sanhuang Yinchi decoction;high mobility group box1(HMGB1);apoptosis;Yinchenhao Tang
Abstract:Objective:To discuss the effect of Yiqi Jianpi Huayu recipe (YQJPHY) combined with 5-fluorouracil(5-FU) on the growth and immune function of subcutaneous transplanted tumor in MFC tumor bearing 615 mice.Method:Twenty-four mice were inoculated subcutaneously to establish the transplanted tumor model of gastric cancer in mice, and then randomly divided into model control group, YQJPHY (20 g·kg-1)group, 5-FU (25 mg·kg-1) group and (YQJPHY+ 5-FU) combined group, with 6 rats in each group. After the last administration, the transplanted tumor, spleen and thymus were stripped completely. The tumor inhibition rate, thymus and spleen index were calculated. Flow cytometry was used to determine the content of myeloid-derived suppressor cells (MDSCs) and its subtype polykaryotype cells (PMN-MDSC), single karyotype cells (M-MDSC) in both peripheral blood and tumor tissue, and macrophages and their M1 type, M2 type, T lymphocyte, B lymphocyte, and natural killer cells (NK cells) in peripheral blood. Expressions of arginase-1(Arg-1) and inducible nitric oxide synthesis (iNOS) gene in tumor tissues were detected by Real-time PCR.Result:Compared with model control group, the weight of mice in YQJPHY group increased, whereas the weight of tumor, the weight of tumor, the index of thymus and spleen decreased in 5-FU group(P<0.05, P<0.01). Compared with 5-FU group, combined group showed gained weight, decreased tumor weight, and increased thymus and spleen index(P<0.05, P<0.01). Compared with model control group, the total MDSCs content in tumor tissue and peripheral blood of the combined group was decreased, and the PMN-MDSC content in 5-FU group and the combined group were decreased(P<0.05, P<0.01). Compared with 5-FU group, the contents of total MDSCs, PMN-MDSC, M-MDSC in combined group were all decreased(P<0.05, P<0.01). Compared with model control group, the number of macrophages and their M1 type in YQJPHY group were higher than those in 5-FU group, while the contents of CD3+ , CD4+ , CD8+ T cell group decreased(P<0.05, P<0.01). Compared with model control group, gene expressions of iNOS, Arg-1 decreased in all groups(P<0.05, P<0.01). Compared with 5-FU group, gene expressions of iNOS, Arg-1 decreased in combined group(P<0.05, P<0.01).Conclusion:YQJPHY can better inhibit the growth of subcutaneous transplanted tumor when combined with 5-FU, and improve immune status after chemotherapy. The mechanism may be related to the decrease of MDSCs content and the increase of T cell and macrophages content.
Abstract:Objective:To investigate the mechanism of Oxymatrine on epithelial-mesenchymal transition mediated by RhoA/Rho-associated kinase(ROCK) signaling pathway to prevent and treat ulcerative colitis(UC) and its related canceration by detecting the changes of ROCK, E-cadherin and transforming growth factor-β(TGF-β)in colon tissues of mice.Method:Totally 48 male Balb/c mice were randomly divided into normal control group, model group, low, medium and high-dose Oxymatrine groups (25, 50, 100 mg·kg-1)and Y-27632 group(10 mg·kg-1), with 8 mice in each group. Mice in control group received distilled water, while all the other mice were treated with 3%dextra sulfate sodium for 7 days to induce the ulcerative colitis model. Since the first day of modeling, Y-27632(10 mg·kg-1)and different doses of Oxymatrine(25, 50, 100 mg·kg-1) were intraperitoneally injectedfor 7 days, and equal volume of PBS was intraperitoneally injected in normal group and model group. Body weight loss, stool consistency and fecal blood loss were observed on a daily basis. On the 8thday, mice were put to death, colon was collected and its length was measured; the scores of disease activity index (DAI) were evaluated; part of the colons were fixed and stained with hematoxylin and eosin (HE) for a histopathological analysis; the ultrastructural changes of mucosa tissue in ulcerative colitis were observed by transmission electron microscope. The expression levels of TGF-β in tissue mucosa were tested by enzyme-linked immunosorbentassay(ELISA). The expression levels of Rho-associated kinase-1, Rho-associated kinase-2, E-cadherin and TGF-β in colon were measured by Western blot and Real-time PCR.Result:Compared with normal group, model group showed the infiltration of a large number of inflammatory cells in mucosa and submucosa, disordered gland arrangement, varying degrees of intestinal mucosal defect and even ulcer formation. Under electron microscopy, microvilli were sparse on the surface of intestinal epithelial cells, the gap between cell junctions was widened, goblet cells were reduced and organelles were swollen. The disease activity index, and the expression levels of ROCK-1 and ROCK-2 proteins in the colonic mucosa of model group were increased(P<0.01), while the colon length and the protein and mRNA contents of E-cadherin, TGF-β were decreased(P<0.01). Compared with model group, there were different degrees of alleviations in pathological manifestationsunder the light and electron microscopy in each treatment group. DAI score and colon length reduction were significantly decreased in each treatment group (P<0.01). The proteins and mRNA expression levels of colonic mucosa ROCK-1 and ROCK-2 of the treatment group were decreased(P<0.01), while the protein and mRNA expression levels of E-caherin and TGF-β were increased(P<0.01), which was statistically significant compared with model group. Compared with middle-dose Oxymatrine group, the ROCK-1 and ROCK-2 protein and mRNA levels were significantly increased in low-dose and high-dose groups(P<0.05, P<0.01), and the TGF-β and E-cadherin protein and mRNA levels were significantly decreased(P<0.01).Conclusion:Oxymatrine may alleviate ulcerative colitis by down-regulating the expression of Rho kinase, up-regulating the expressions of E-cadherin and TGF-β, inducing the apoptosis of intestinal epithelial cells, and mediating epithelial-mesenchymal transition.
Abstract:Objective:To investigate the effect of ginsenoside 20(S), 25-epoxydammarane-3β, 12β, 24α-triol (24-OH-panaxadiol, 24-OH-PD) on inhibiting proliferation and inducing apoptosis of tumor cells, and explore its mechanism of action.Method:The inhibitory effect of 24-OH-PD (12.5, 25, 50, 100 mg·L-1) on proliferation of CCRF-CEM, M14, MD-MBA-231 and Jeko-1 cells with different treatment periods (24, 48, 72 h) was evaluated by methylthiazolyldiphenyl-tetrazolium bromide (MTT)assay and CellTiter Glo® test, and the results were then compared with 20(R)-Rg3 and 20(S)-Rh2.Flow cytometry was used to detect cell apoptosis caused by 24-OH-PD. Besides, the potential anticancer mechanism was studied by docking analysis with 40 cancer related proteins and 24-OH-PD by using drug design platform Schrodinger Maestro 6.7 Software.Result:24-OH-PD inhibited the proliferation of all the 4 cancer cell lines significantly in a time and dosage dependent manner. The IC50 value of 24-OH-PD on CCRF-CEM, Jeko-1, M14, and MD-MBA-231 cell lines was 25.36, 39.29, 21.74, and 19.35 mg·L-1, respectively, similar to 20(S)-Rh2 (IC50 23.35, 65.79, 18.95, 19.67 mg·L-1) and much better than 20(R)-Rg3 (only effective for Jeko-1 cells, IC50 49.5 mg·L-1). Annexin V/PI double staining experiment showed that 24-OH-PD could also induce apoptosis of the 4 kinds of cancer cells (P<0.05) in a dose-dependent manner. In the molecular docking test, 24-OH-PD was docked successfully with 11 tumor related proteins, including purine nucleoside phosphorylase (PNP), protein tyrosine kinase, protein kinase C (PKC), B-cell lymphoma-2 (Bcl-2), B-cell lymphoma-xl (Bcl-xl) and Caspase-8 et al, which demonstrated that the anti-tumor effect of 24-OH-PD may be related to the direct effects on these proteins.Conclusion:24-OH-PD could inhibit cell proliferation and induce apoptosis for CCRF-CEM, M14, MD-MBA-231 and Jeko-1 cell lines, which may through directly acting on Bcl-2, Bcl-xl, and other proteins.
Abstract:Objective:To investigate the intervention effect of panax notoginseng saponins (PNS) on epithelial-mesenchymal transition (EMT) of rat renal tubular epithelial cells (NRK-52E) induced by transforming growth factor-β1(TGF-β1), and analyze the mechanism based on the silent information regulation 2 homolog 1(SIRT1)/TGF-β1/Smad signaling pathway.Method:NRK-52E were cultured in DMEM medium with 10%fetal bovine serum, and divided into normal control group, TGF-β1 group (5 μg·L-1), resveratrol (RSV) group (50 mg·L-1), EX527 group (10 μmol·L-1), Panax notoginseng saponins (PNS) group (100 mg·L-1), and EX527+ PNS group (10 μmol·L-1+ 100 mg·L-1). Then cells were collected after drug intervention for 48 h. The expressions of α-SMA, E-cadherin, SIRT1, TGF-β1, Smad3, Smad4 mRNA in each group were detected by Real-time PCR. The protein expressions of α-SMA, E-cadherin, SIRT1 and TGF-β1 were detected by Western blot.Result:Compared with normal group, mRNA and protein expressions of α-SMA increased obviously(P<0.05), but E-cadherin decreased significantly(P<0.01)in TGF-β1 group. Compared with TGF-β1 group, mRNA and protein expressions of α-SMA decreased significantly(P<0.01), while E-cadherin increased(P<0.01)in resveratrol and PNS groups, and EMT was inhibited. Meanwhile, mRNA and protein expressions of SIRT1 increased significantly(P<0.01), while mRNA expressions of TGF-β1, Smad3, and Smad4 decreased(P<0.01). Under the intervention of SIRT1 blocker EX527, PNS could not play a significant inhibitory effect on the cells.Conclusion:PNS can prevent the occurrence of EMT of renal tubular epithelial cells induced by TGF-β1, and the mechanism may be related to active SIRT1 to inhibit TGF-β1/Smad pathway.
Abstract:Objective:To observe effect of Xiaoai Jiedu formula combined with fluorouracil (5-FU)+ epirubicin (EPI)+ cyclophosphamide (CTX) (FEC) chemotherapy regimen on immune function, tumor index, traditional Chinese medicine (TCM) symptom scale score and adverse reactions of patients with breast cancer.Method:A total of 60 patients with breast cancer were randomly divided into study group and control group. FEC (CTX 0.6 g·m-2, d1, EPI 100 mg·m-2, d1, 5-FU 0.5 g·m-2, d1) regimen was used in study group, and Xiaoai Jiedu formula recipe was used for two consecutive cycles. FEC regimen was used in control group only. After 2 cycles, immunological changes, tumor index, TCM symptom score and adverse reactions were analyzed and compared between two groups.Result:There was not significant difference in immune indexes between two groups before treatment, but statistically significant differences after treatment (P<0.05). There was no statistically significant difference in levels of carcinoembryonic antigen(CEA) and carbohydrate antigen153(CA153) between two groups before treatment; CEA and CA153 levels significantly decreased in both groups, particularly in study group (P<0.05). There was no statistically significant difference in scores of fatigue, hot flashes and night sweats, irritability, dry mouth, appetite and sleep between two groups before treatment. The scores in study group were significantly lower than those in control group after treatment (P<0.05). The incidence of menstrual disorder, nausea, vomiting and constipation in study group was lower than that in control group (P<0.05).Conclusion:Xiaoai Jiedu formula combined with FEC chemotherapy can improve clinical efficacy and alleviate clinical symptoms of patients.
Abstract:Objective:To make statistics on the annual frequency of patients with eczema by traditional Chinese medicine (TCM) physique, syndrome differentiation, and western medicine staging based on questionnaire survey, in order to infer the distribution and characteristics of the annual frequency by different TCM physique, syndrome differentiation and western medicine stage, and provide new ideas and new methods for the prevention and treatment of Eczema.Method:According to the Dermatovenerology of Traditional Chinese Medicine edited by QU Xing, and Clinical Diagnosis and Treatment of Chinese Medicine for Dermatovenerology edited by CHEN Da-can, and Traditional Chinese Medicine for Dermology edited by YU Wen-qiu, and Physique Classification and Patient Self-Testing Table formulated by China Association of Chinese Medicine and Professor WANG Qi's nine categories of physique, the TCM Physique Classification and Patient Self-Testing Table for eczema patients and the Syndrome Differentiation and Classification Table for Eczema Patients were formulated. General conditions of 482 cases of eczema patients treated at Tianjin Academy of Traditional TCM Affiliated Hospital and their types of TCM physique, TCM syndrome differentiation, western medicine staging and annual frequency were surveyed.Result:There were significant differences in the annual frequency of patients with different physical constitutions. By single physique, Tanshi, Shire and Qiyu had more frequent occurrences every year, and Pinghe had the lowest annual frequency. There were differences in the annual occurrences among cases with different TCM syndrome types. Shire syndrome patients have more frequent annual occurrences than other types of eczema patients. There were significant differences in the annual occurrences among cases of different western medicine staging, and the annual occurrences of acute eczema were more than those of subacute and chronic eczema.Conclusion:The annual occurrences of patients with Tanshi, Shire and Qiyu physique were higher than those of other physiques. There are fewer outpatients with Pinghe physique than other physiques. The annual occurrences of Shire syndrome patients are higher than that of other types of eczema patients. The annual occurrences of acute eczema are more than those of subacute and chronic eczema. The annual occurrences of chronic eczema are less than acute and subacute eczema.
Keywords:eczema;traditional Chinese medicine (TCM) physique;syndrome differentiation;western medicine stage;annual frequency
Abstract:Objective:To observe the efficacy of Xiaoxuan Zhiyun tablet in treating cervical vertigo with syndrome of phlegm and blood stasis resistance and its effect on oxidative stress and serum levels of endothelin-1(ET-1), 5-hydroxytryptamine (5-HT), and nitric oxide (NO).Method:One hundred eighteen cervical vertigo cases were selected and randomly divided into control group (59 cases) and treatment group (59 cases) by random number table. Control group was given bone setting combined with acupuncture. In addition to the therapy of control group, treatment group was given Xiaoxuan Zhiyun tablet (1 dose/day, tid). Both groups were treated for 3 weeks. Scores of cervical vertigo symptom and function assessment scale (ESCV) and vertigo disorder scale (DHI), blood velocity of vertebrobasilar artery, life quality and efficacy were compared between both groups. Serum levels of superoxide dismutase (SOD), malondialdehyde (MDA), ET-1, 5-HT and NO were detected in both groups.Result:After treatment, ESCV score of treatment group was obviously higher, while DHI score was lower than control group (P<0.01). Blood flow velocity of left vertebral artery, right vertebral artery and basilar artery in treatment group were obviously higher than those of control group after treatment (P<0.01). Total clinical efficacy of treatment group wsa 98.25%, which was superior to 83.93% of control group (P<0.05). After treatment, SF-36 index score was remarkably higher than that of control group (P<0.01). Serum levels of SOD and NO of treatment group were higher, while MDA, ET-1, 5-HT were lower than control group (P<0.01).Conclusion:In addition to the routine therapy, the efficacy of Xiaoxuan Zhiyun tablet in treating cervical vertigo is significant, which may be related with regulation of oxidative stress and serum levels of ET-1, 5-HT and NO.
Keywords:Xiaoxuan Zhiyun tablet;cervical vertigo;syndrome of phlegm and blood stasis resistance;oxidative stress
Abstract:Objective:To observe the clinical efficacy of Shugan granule on perimenopausal depressive disorder (stagnation of liver Qi and Qi stagnation) and its effect on sex hormone and inflammatory factors.Method:One hundred and twenty patients were randomly divided into control group (60 cases) and observation group (60 cases) by random number table. Both groups got Tibolone tablets, 2.5 mg/time, 1 time/day. Patients in control group got flupentixol and melitracen tablets once every morning and noon, 1 tablet/time. In addition to the therapy of control group, patients in observation group got Shugan granule, 1 pack/time, 2 times/days. And a course of treatment was 8 weeks. Main indexes were graded by Hamilton depression scale (HAMD-17), self-rating depression scale (SDS) and Eysenck Personality Questionnaire (EPQ). And secondary indexes were scored by self-rating anxiety scale (SAS), stagnation of liver Qi and Qi stagnation, improved kupperman (KI), levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), 5-serotonin (5-HT), brain derived neurotrophic factor (BDNF), interleukin-1β (IL-1β), IL-6 and tumor necrosis factor-α (TNF-α) before and after treatment.Result:By rank sum test, the clinical efficacy on depressed and perimenopausal syndrome in observation groups was better than that in control group (Z=2.038, P<0.05, Z=2.316, P<0.05). Scores of HAMD-17, SDS, EPQ-N, SAS, KI and stagnation of liver qi and qi stagnation were lower than those in control group (P<0.01), whereas score of EPQ-E was higher than that in control group (P<0.01). And levels of FSH, LH, IL-1β, IL-6 and TNF-α were lower than those in control groups (P<0.01). Levels of E2, BDNF and 5-HT were higher than those in control groups (P<0.01).Conclusion:Shugan granule can relieve depression, inquietude and symptoms caused by perimenopausal syndrome, regulate sex hormone, 5-HT, BDNF and inflammatory reaction, with a better clinical effect than pure western medicine treatment.
Abstract:Objective:In the study of urine metabolomics of rats, necessary antiseptic measures should be taken for collection of urine samples, the effect of several antiseptic measures on the endogenous metabolites in urine was studied.Method:The urine samples of rats were collected on ice, sodium azide was added, and both of them were used together to prevent corrosion. Differences of antiseptic measures were analyzed by nuclear magnetic resonance (NMR) metabolomics. Result:The results of NMR metabolomics showed that sodium azide+ ice group and ice group had many overlaps, but they clearly separated with the control group and sodium azide group; sodium azide group and the control group had a small part overlap, but there was a tendency of separation. The antiseptic effect of sodium azide+ ice group and ice group was similar; compared with control group, valine, betaine and hippuricacid in these two groups increased, but the alanine and 2-ketoglutaric acid decreased. Conclusion:In the study of rat urine metabolomics, low temperature antiseptic measures must be taken when urine samples are collected, and the addition of sodium azide can improve the antiseptic effect slightly under protective conditions.
Keywords:urine;endogenous metabolites;nuclear magnetic resonance metabolomics;sodium azide;ice;antiseptic measures;protective conditions
Abstract:Objective:To study the serum pharmacochemistry of Periploca forrestii rhizomes, and to investigate the pharmacological material basis of extract of P. forrestii rhizomes in rats.Method:Rapid identification of constituents absorbed into blood was carried out by UPLC-Q-TOF-MS, according to retention time, accurate relative molecular mass and standard substance comparison, these constituents were identified and speculated by Data Analysis, Metabolite Detect and other softwares, then preliminary determination of constituents absorbed into blood of rats after oral administration of extract of P. forrestii rhizomes was investigated.Result:Totally 17 constituents absorbed into blood were detected in serum, ten of them were prototype constituents and the other were metabolites. Seven of the prototypes were identified as 5-O-caffeoylquinic acid, 4-O-caffeoylquinic acid, 3-O-caffeoylquinic acid, 3, 4-di-O-caffeoylquinic acid, 3, 5-di-O-caffeoylquinic acid, 4, 5-di-O-caffeoylquinic acid and periplocin. Conclusion:These constituents absorbed into blood may be substances that act directly in vivo of P. forrestii rhizomes, and it is helpful to clarify pharmacological material basis and mechanism of this herb.
Keywords:Periploca forrestii rhizomes;serum pharmacochemistry;caffeoylquinic acid derivates;pharmacodynamic material basis;periplocin;extract;metabolic pathways
Abstract:Objective:To explore the difference of effect of raw and wine-processed products of Rhei Radix et Rhizoma extract on brain protection of rats with intracerebral hemorrhage(ICH) and the connotation of the theory of "wine processing could promote efficacy" of Rhei Radix et Rhizoma.Method:Wistar male rats were used to establish the ICH model, extracts of raw and wine-processed products of Rhei Radix et Rhizoma were used for intervention, and these rats were randomly divided into 6 groups(blank group, sham-operated group, model group, raw Rhei Radix et Rhizoma group, wine-processed Rhei Radix et Rhizoma group and Angong Niuhuangwan group). Then the apoptosis of hippocampal neurons was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL) assay. The expression of zonula occludens-1(ZO-1) and vascular endothelial cadherin-2(VE-cadherin-2) in the hemorrhagic brain tissue was detected by immunohistochemical assay. The expression of reduced form of nicotinamide-adenine dinucleotide phosphate(NADPH) oxidase 2(NOX2) and tumor necrosis factor-α stimulated gene-6(TSG-6) in the hemorrhagic brain tissue was detected by Western blot. The content of glutathione(GSH) in the serum of ICH rats was detected by automatic enzyme-linked immunosorbent assay systems. Through these above methods, we investigated the differences between raw products and wine-processed products on brain protection of ICH rats. Result:Compared with the sham-operated group, the apoptosis index of the hippocampal neurons in the model group were increased significantly(P<0.01), the expression of ZO-1 was decreased significantly(P<0.01), the expression of NOX2 protein was increased significantly(P<0.01), the expression of TSG-6 protein was decreased significantly(P<0.01), the content of GSH was also decreased significantly(P<0.01). Compared with the model group, raw and wine-processed products of Rhei Radix et Rhizoma could reduce the apoptosis index of hippocampal neurons(P<0.01), increase the expression of ZO-1(P<0.01) and VE-cadherin-2 protein(P<0.05), decrease the expression of NOX2(P<0.05, P<0.01), increase the expression of TSG-6 protein(P<0.05) and increase the content of GSH(P<0.01) in ICH rats.Conclusion:The raw and wine-processed products of Rhei Radix et Rhizoma all have brain protective effect on ICH rat model, but the protective effect of wine-processed products is slightly better than that of raw products. The result of this study provides experimental basis for exploring the theory of "wine processing could promote efficacy" of this herb.
Keywords:wine processing could promote efficacy;intracerebral hemorrhage;cerebral protection;blood-brain barrier;Rhei Radix et Rhizoma;extract;wine-processed products
Abstract:Objective:To establish the fingerprint of standard decoction of Citri Reticulatae Pericarpium and evaluate its quality.Method:According to the preparation conditions of the standard decoction, 15 batches of standard decoction of Citri Reticulatae Pericarpium were prepared. HPLC was employed to determine the content of hesperidin in this standard decoction. Ultraviolet spectroscopy(UV) and infrared spectroscopy(IR) were used to establish the fingerprint of standard decoction of Citri Reticulatae Pericarpium. The correlation coefficient method and double index sequence analysis method were used to compare and analyze the spectra of different batches of this standard decoction. Result:The content of hesperidin in 15 batches of this standard decoction were 0. 82%-2. 60%, and the measured value of dry extract rate was 32. 02%-46. 11%. Compared with ultraviolet and infrared control fingerprint, the fingerprint similarities of the standard decoction of each batch were >0. 897 and >0. 942, respectively. The double index analysis results showed that the common peak ratio was more than 62. 50%, variation peak ratio was less than 46. 67%. Conclusion:The quality evaluation method established in this study can be used for systematic evaluation of standard decoction of Citri Reticulatae Pericarpium, and it can provide theoretical reference for the formulation of quality standard of Citri Reticulatae Pericarpium dispensing granules and other related preparations.
Abstract:Objective:To study the dry extract rate, determination and transfer rate of maker compounds, fingerprint and others of standard decoction of Chrysanthemi Indici Flos and provide basic data for the preparation of this standard decoction and its dispensing granules by establishing 15 batches of standard decoction of Chrysanthemi Indici Flos from 5 different places.Method:The standard decoction of Chrysanthemi Indici Flos was prepared based on the traditional decoction process, the content of linarin was determined by UPLC-DAD, the transfer rate of this composition was calculated, the fingerprint was drawn, the extract powder was prepared by vacuum drying, and the dry extract rate was calculated.Result:The concentration of linarin in 15 batches of standard decoction of Chrysanthemi Indici Flos was 0.19-0.74 g·L-1, the transfer rate of linarin was 21.95%-66.23%, its average transfer rate was 37.12% with RSD of 11.8%, the pH value was 5.1-5.5, the range of dry extract rate was 24.7%-32.5%, the average dry extract rate was 27.87% with RSD of 2.4%.There were 9 major common peaks in the fingerprint and 2 peaks(No. 2 and No. 9) were confirmed, such as chlorogenic acid and linarin.Conclusion:The preparation method in this research conforms to the traditional decoction method and is stable and feasible. It can be used for the preparation and quality evaluation of standard decoction of Chrysanthemi Indici Flos.
Keywords:standard decoction of Chrysanthemi Indici Flos;standard decoction;fingerprint;dispensing granules;classic formula;linarin;chlorogenic acid
Abstract:Objective:To characterize and compare the chemical information of four extracts of Qingre Chushi (QRCS) decoction by liquid chromatography-mass spectrometry (LC-MS), and combine the chemical information of the four extracts with their results of anti-inflammatory effect for a multivariate statistical analysis, in order to identify the compounds directly relating to the anti-inflammatory effects of QRCS decoction.Method:Four extracts of QRCS decoction were characterized by UPLC-Q-TOF-MS: ①ethanol extract+ water extract, ② ethanol extract+ supernatant after water extraction and alcohol precipitation, ③ ethanol extract+ precipitation after water extraction and alcohol precipitation, and ④ standard decoction. On the basis of the results of inhibition of the four above extracts on xylene-induced ear swelling in mice, multivariate statistical analysis [principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA)] were carried out to lock the chromatographic peaks with significant differences between group ① (the best pharmacological action group) and group ④ (standard decoction group). According to the accuracy of quasi-molecular ion and fragment ion data, and the reference materials and literature data, those chromatographic peaks were identified.Result:PCA could cluster the four extracts of QRCS decoction, and the differences between groups was reflected in the distance between groups. Group ④ (standard decoction) had the most significant differences with the other three groups, especially in the first principal component; group ① (ethanol extract+ water extract), group ② (ethanol extract+ supernatant after water extraction and ethanol precipitation) and group ③ (ethanol extract+ precipitation after water extraction and ethanol precipitation) had certain differences in the second principal component. OPLS-DA was used to compare group ① (the best pharmacological action group) and group ④ (standard decoction group). Eleven chromatographic peaks with great contribution and high reliability to group differences, were identified as gentiopicrin, skimmin, baicalin, baicalin isomer, wogonoside, 5, 6, 7-trihydroxy-8-methoxyflavone-7-O-glucurodonaldehyde, 5, 6-dihydroxy-6, 8, 2′, 3′-tetramethoxyflavone, salicin-6-C-arabinose-8-C-glucoside, plantamajoside and glycyrrhizic acid.Conclusion:In the mode of pectrum-effect combination, this study explores and identifies compounds relating to the anti-inflammatory effect of QRCS decoction, so as to provide the basis for screening the extraction and purification process and optimizing the formulation of preparation of Qingre Chushi decoction.
Keywords:UPLC-Q-TOF-MS;Qingre Chushi decoction;principal component analysis;orthogonal partial least squares-discriminant analysis
Abstract:Objective:To systemically study the chemical constituents of n-butanol fraction from Lysimachia capillipes.Method:The whole plant of L. capillipes was crushed into power, extracted by 70%methanol, concentrated under reduced pressure, and then its n-butanol extract was obtained by fractional extraction. The compounds from n-butanol fraction were isolated and purified by macroporous resin column chromatography, medium pressure ODS, silica gel, Sephadex LH-20 and preparative HPLC. Their structures were identified on the basis of spectral analysis and comparison with literature data.Result:Fifteen compounds including 6 saponins and 9 flavonoid glycosides were isolated from L. capillipes, and were identified as ascapilliposide B(1) and capilliposide C(2), kaempferol-3-O-β-D-xylopyranosyl(1→3)-[4-O-E-p-coumaroyl-α-L-rhamnopyranosyl(1→2)][β-D-glucopyranosyl(1→6)]-β-D-galactopyranoside-7-O-α-L-rhamnopyranoside(3), kaempferol-3-O-{[β-D-xylopyranosyl(1→3)-α-L-rhamnopyranosyl(1→6)][α-L-rhamnopyranosyl(1→2)]}-β-D-3-trans-p-coumaroylgalactopyranoside (4), capilliposide K (5), 3β-O-{α-L-rhamnopyranosyl-(1→2)-O-β-D-glucopyranosyl-(1→4)-[O-β-D-glucopyranosyl-(1→2)]-α-L-arabinopyranosyl)}-16α-hydroxyolean-28, 13β-olide (6), capilliposide I(7), quercetin-3-O-(2″, 6″-di-O-α-rhamnopyranosyl)-β-galactopyranoside(8), kaempferol-3-O-{[β-D-xylopyranosyl(1→3)-α-L-rhamnopyranosyl(1→6)][α-L-rhamnopyranosyl-(1→2)]}-β-D-galactopyranoside(9), kaempferol-3-O-[2-glucopyranosyl(1→3)rhamnopyranosyl-6-rhamnopyranosyl]-β-D-galactopyranoside(10), kaempferol-3-O-α-L-rhamnopyranosy-(1→2)-[α-L-rhamnopyranosy-(1→6)]-β-D-galactopyranoside(11), capilliposide I(12), kaempferol-3-O-{(β-D-glucopyranosyl-(1→3)-[4-O-(E-p-coumaroyl)]-α-L-rhamnopyranosyl-(1→6)-(β-D-galactopyranoside)}-7-O-α-L-rhamnopyranoside (13), kaempferol-3-O-{[β-D-glucopyranosyl(1→3)]-4-O-(E-p-coumaroyl)}-α-L-rhamnopyranosyl(1→6)-β-D-glucopyranoside-7-O(4-O-acetyl)-α-L-rhamnopyranoside (14), and (3β, 20S, 23S, 24R)-3, 20, 23, 24, 25, 29-hexahydroxydammaran-21-oic acid-21, 23-lactone 3-O-β-D-glucopyranosyl-(1→6)-β-D-gluco-pyranoside(15).Conclusion:The compounds 3, 4, 6, 9, 10, 13-15 were isolated from this plant for the first time.
Abstract:Objective:To study the antibacterial activity and chemical constituents of Sojae Semen Praeparatum as a traditional Chinese medicine.Method:Sojae Semen Praeparatum was extracted with 80%ethanol, and the extracts were suspended in an appropriate amount of distilled water and extracted with petroleum ether, ethyl acetate and water saturated n-butanol in turn. The antibacterial activity in vitro was used for screening the active fractions. Using activity-guided-isolation, the active fractions were isolated and purified by silica gel column chromatography and preparative HPLC and other methods. Their structures were identified by physicochemical properties and spectroscopic analysis, such as 1H-NMR, 13C-NMR and MS.Result:The ethyl acetate fraction of 80%ethanol extract from Sojae Semen Praeparatum showed a stronger activity and was studied further. Eighteen compounds were isolated, and eleven compounds were identified as daidzein (1), daidzin (2), genistein (3), genistin (4), glycitein (5), glycitin (6), daucosterol (7), thymine (8), adenine (9), uracil (10), uridine (11).Conclusion:The ethyl acetate fraction of 80%ethanol extract from Sojae Semen Praeparatum shows an antibacterial activity. Compounds 1-6 are isoflavones. Compounds 7-11 are isolated from Sojae Semen Praeparatum for the first time.
Abstract:Objective:To optimize the entropy TOPSIS model to evaluate the quality of Hemerocallis Flava from different regions, in order to provide a new evaluation method for the quality control of traditional Chinese medicine.Method:The entropy weight TOPSIS model optimized by analytic hierarchy process(AHP) method was used to analyze the quality of Hemerocallis Flava from 14 different regions, and a comprehensive evaluation index system for the quality of Hemerocallis Flava, which covered 3 layers (target layer, decision layer and index layer), and 10 indexes (corolla of Hemerocallis Flavathe's bud, pistils, stamens, peduncle length, extract, total ash, quercetin, β-rhamnocitrin, kaempferol, sitosterin) was established.Result:Qingyang showed the best quality of Hemerocallis Flava, which was followed by Weinan, and the lowest quality was found in Datong, Shanxi, and Xiaowan village, Sichuan. The results were consistent with the evaluation results of traditional empirical identification, suggesting the successful modeling. The contents of β-rhamnocitrin and kaempferol in Qingyang were 1.72 times and 2.74 times of those of Xiaowan village, Sichuan. There was no significant difference in other active ingredients from different regions. It was suggested that quercetin and kaempferol could be used as the identification and quality evaluation indexes of cauliflower.Conclusion:The entropy TOPSIS model based on the AHP method is clear, simple to use and easy to calculate, with distinct evaluation indexes. It is a practical, quick and effective comprehensive evaluation method for multi-objective decision analysis.
Keywords:analytic hierarchy process method;entropy TOPSIS;comprehensive evaluation;Hemerocallis Flava
Abstract:Objective:To establish a gas chromatography(GC) fingerprints analysis method of farmed musk, natural musk and artificial musk for effective control and scientific evaluation of the quality of musk.Method:The sample was split in the 250 ℃ injection port, with 1∶1 split ratio, and separated on a Capillary column(325 ℃, 0.25 mm×30 m, 0.25 μm). Nitrogen was used as a carrier gas, the flow rate was 1 mL·min-1. The detector temperature was 280 ℃. The temperature program was 80 ℃ for 2 min, increase by 5 ℃·min-1 to 160 ℃, and by 1 ℃·min-1 to 200 ℃ for 20 min, then 5 ℃·min-1 to 260 ℃ for 20 min. GC of the 30 batches of farmed musk, 24 batches of natural musk and 8 batches of artificial musk were analyzed by adopting temperature programmer. The similarity evaluation system for chromatographic fingerprint of TCM(2004A)was used to evaluate the similarity of these chromatograms. The fingerprint peaks were identified by using GC-MS combination technology. The ion source temperature was set at 230 ℃. The quadrupole temperature was set at 150 ℃. The GC-MS interface temperature was set at 280 ℃. Electron impact(EI)was used as the ionization source, and the electron energy was set at 70 eV. The scanning scope was m/z 30-550.Result:The GC fingerprints of different kind of musk were established, and the similarities all conformed to regulations. Totally 17 common compounds in farmed musk, 10 common compounds in natural musk, and 8 common compounds in artificial musk were determined and compared.Conclusion:This method features a high precision, reproducibility, and stability. The established fingerprints could be used for the overall quality evaluation for musk.
Abstract:Objective:To provide evidence and lay a foundation for quality control and resources utilization of Kochiae Fructus.Method:A total of 23 samples of Kochiae Fructus were collected from the main distribution areas in China. Momordin Ⅰc content was determined via HPLC-ELSD method, and its correlation with growth traits and habitat factors was analyzed using SPSS 23.0 software, as well as the principle component and clustering analysis.Result:Momordin Ⅰc content had a benign linear relationship with HPLC-ELSD peak area within the range between 29.8 mg·L-1 and 178.8 mg·L-1 (R2=0.999 8), with the quantitation limit at 5.79 mg·L-1 and the detection limit at 2.36 mg·L-1. Significant differences (P<0.01) were found in momordin Ⅰc contents among 23 Kochiae Fructus germplasms. According to momordin Ⅰc content, all samples tested met the requirements in the Chinese Pharmacopeia published in 2015.Origin, growth traits, and habitat factors could impact momordin Ⅰc content accumulation in Kochiae Fructus. Moreover, the content had a negative correlation with plant height, stem diameter, leaf length, leaf width, fruit diameter and longitude, and a positive correlation with ratio of length to width of leaves, altitude and latitude. Those from Shanxi, Jilin, and Shandong provinces had a higher momordin Ⅰc content. The characters tested could be simplified to four principle components, and their cumulative contribution rate was 95.831%. Kochiae Fructus samples tested could be divided into four groups, and those from group Ⅰ had superior growth traits, and those from group Ⅲ had higher momordin Ⅰc content.Conclusion:The optimized HPLC-ELSD method for determining momordin Ⅰc content of Kochiae Fructus is convenient, accurate and reproducible, and could be used to control the herbs quality. Those cultivated in a lower longitude, higher altitude, and higher latitude region could accumulate more momordin Ⅰc content. More attention shall be paid to germplasm in Shanxi and Zhejiang with superior resources screening of Kochiae Fructus.
Abstract:Through analysis of factors affecting toxicity of traditional Chinese medicine (TCM), relationship between toxicity of TCM and physical constitution was explored, which provided a new idea for preventing occurrence of toxicity of TCM. Efforts were made to summarize causes of toxicity of TCM, and analyze relationship between physical constitution and toxicity of TCM. following aspects were taken into account: ①main reason for people to use TCM is disease, which is pathological state of human beings. The pathological state may have more demands for corresponding TCM. The TCM rule of "disease-based treatment" is an important manifestation for correlating drugs with syndromes. Then how can normal animals reflect human toxicity under pathological conditions. ②People with pathological conditions, especially with a long disease course, may have abnormal liver and kidney functions. Then how to distinguish cause of toxicity of TCM in liver and kidney dysfunctions.③The long-term disease state can also lead to disorder of intestinal flora of body. Then how is impact of interaction between intestinal flora and TCM reflected in toxicity of TCM.④Although emotional and psychological factors account for a small proportion of toxicity of TCM, it still has an effect.⑤Although humans and animals have a high degree of similarity, there are still species differences (such as croton). Then how is human clinical toxicity consistent with animal research toxicity. The efficacy is related to expectation, while toxicity is related to background. At present, toxicity of TCM is more caused by factors of TCM (such as variety, quality, processing), and factors of TCM application (like compatibility, drug correspondence, dosage, course of treatment), which are external causes of toxicity of TCM. physiological condition, pathological state, emotional and psychological factors, and intestinal flora of body are internal factors. The physical constitution factors are internal factors of toxicity of TCM, and external factors play a role based on internal factors. At the same time, there are essential differences between humans and animals. Animal toxicological experiments are only for clinical reference. By analyzing concepts related to toxicity of TCM and physical constitution factors, modern researches on toxic effects of TCM are also promoted accordingly, and TCM toxicity theory is also improved, which provides a scientific basis for safe and rational application of TCM in clinical practice. Of course, research on animal models under pathological conditions shall be strengthened to promote study of toxicity of TCMs, so as to deepen people's understanding of toxicity of TCMs.
Keywords:traditional Chinese medicine toxicity;organism factors;pathological state;animal model
Abstract:Endometriosis is a multiple disease that afflicts the health of women at childbearing age, and its incidence rate has been increasing year by year, furthermore, there has been a trend to be younger. At present, the pathogenesis of endometriosis has been not expounded completely, its cure rate is not high with high recurrence rate. In recent years, studies have shown that the human is a commensal body composed of a large number of microorganisms, and especially the microorganisms in the intestinal are closely related to the health of the body. Based on the previous studies on endometriosis, this paper proposes that its pathogenesis may be related to intestinal microbiological disorder, and aims to provide new ideas for the treatment of endometriosis.
Keywords:endometriosis;intestinal flora imbalance;bacterial endotoxin;epithelial-mesenchymal transition;immunotolerance;lipopolysaccharide;pelvic mass
Abstract:Polycystic ovary syndrome (PCOS) is a common gynecological reproductive endocrine and metabolic disorder disease. The clinical manifestations of PCOS are diversified, with and no ovulation, insulin resistance, hyperinsulinemia and polycystic ovaries as the main pathological and physiological changes, which can lead to non-ovulation infertility and ovulatory dysfunction infertility, cause complications, such as amenorrhea, diabetes, lipid metabolism and glucose metabolism disorder, and endanger the long-term health of the body. Traditional Chinese medicine (TCM) does not have the name of this disease, and classifies PCOS to the category of " amenorrhea" , "infertility" and "abdominal mass kidney" according to its clinical manifestations. Compared with western medicine, TCM has a unique view in treating the disease, and believes that the disorders of liver, kidney and spleen functions jointly affect the occurrence and development of the disease. According to the literature review, kidney-deficiency syndrome is the main syndrome type of this disease, the kidney-tonifying method has achieved a good effect in clinic. Besides, the drug use is flexible and diversified, and both primary and secondary symptoms are taken into consideration. However, in the clinical application process, the application of kidney-tonifying method in PCOS has been affected by the thinking mode of "kidney-tonifying medicine for male diseases" to some extent. In view of this, the therapeutic effect of kidney-tonifying drugs in the treatment of PCOS is preliminarily demonstrated from several aspects of the relationship between kidney deficiency and reproduction, pathogenesis of PCOS, clinical research and experimental research of PCOS, in the expectation of providing theoretical and practical supports for kidney-tonifying therapy for PCOS and other gynecological diseases.
Keywords:polycystic ovary syndrome (PCOS);deficiency of the kidney;reinforcing kidney traditional Chinese medicine (TCM);etiology and pathogenesis
Abstract:Ischemic heart disease is one of the most deadly diseases in the world, and new therapies and preventive measures are urgently needed. In general, cardiomyocytes rely on adenosine triphosphate(ATP) produced by mitochondrial oxidative phosphorylation to maintain their systolic and ion pump functions. Autophagy is a procedural degradation mechanism widely present in eukaryotic cells. It is a self-defense mechanism and self-repair process of the body tissues. It is also a way of apoptosis and a basic phenomenon to maintain the energy balance of human cells. Mitochondrial autophagy is a type of selective autophagy in cells. In fact, damaged mitochondria selectively remove damaged proteins and organelles with autophagy to maintain intracellular homeostasis. Mitochondrial autophagy is important for maintaining the homeostasis of cardiomyocytes. With the deepening of modern biological research, more and more traditional Chinese medicines(TCM) or their extracts have been proven to alleviate myocardial cell damage after ischemia/reperfusion through autophagy or regulation of mitochondrial function. This further inspires TCM workers to find effective treatment measures by targeting mitochondria. Under the above background, this paper reviews the effects of mitochondrial autophagy on ischemic heart disease and the intervention studies of TCM in this field.
Keywords:ischemic heart disease;mitochondria;autophagy;traditional Chinese medicine(TCM)
Abstract:Cinnamomi Ramulus and Cinnamomi Cortex are widely used to treat paralysis in traditional Chinese medicine (TCM). There are numerous and complicated relative records in ancient literatures. Doctors often use Cinnamomi Ramulus to dispel wind and cold, remove blood stasis and combine with warm-natured and heat-natured herbs to treat excess paralysis and early-stage paralysis. And Cinnamorni Cortex is used to warm and invigorate kidney Yang and combine with warm-benefiting herbs to treat deficiency paralysis and chronicle paralysis. However, modern pharmaceutical studies reported that their active substances are almost the same. The active substances in Cinnamomi Cortex are more than those in Cinnamomi Ramulus. The mechanisms of treating paralysis include: suppressing inflammation and regulating immunity by down-regulating nuclear factors(NF)-κB, mitogen activated protein kinase(MAPK), Janus kinase-signal transducers/activators of transcription(JAK/STAT) signaling pathways, regulating cell proliferation by inhibiting the proliferation of fibroblasts, osteoclasts and bone marrow mesenchymal stem cells and promoting the proliferation of osteoblast, resisting oxidation by scavenging oxygen free radicals, regulating pain by mediating TRPA1 and TRPV1, and enhancing substance metabolism and losing weight by regulating the secretion of intestinal hormones (Ghrelin, GLP-1) and improving insulin resistance. The main active ingredient Cinnamaldehyde is unstable in vivo and easily oxidized to cinnamic acid. The toxicity of the two medicines and their components are relatively low. This paper reviews and analyses relative records in ancient literatures, traditional Chinese medicine cognition of their effects in treating paralysis, the achievements and problems of chemical, pharmacological, pharmacokinetic and toxicological researches in recent years, with the aim to provide theoretical basis for further research and application.
Keywords:paralysis;Cinnamomi Ramulus;Cinnamomi Cortex;cinnamaldehyde;modern pharmaceutical Research
Abstract:Known as "the king of the five grains" , tartary buckwheat (Fagopyrum tataricum) is a medicinal and edible plant with both nutritional and healthcare functions. It contains rich flavonoids, such as rutin, with balanced amino acid composition and multiple effects, like blood fat-lowering, blood giucose-lowering, blood pressure-lowering, anti-oxidation, anti-aging, anti-cancer, anti-cancer, and microcirculation-improving. Transcription factors play important roles in plant growth and development by regulating gene expressions. MYB family is one of the largest transcription factor families in plant, contains the MYB domain, and can be divided into four subfamilies: 1R-MYB, R2R3-MYB, 3R-MYB, 4R-MYB. This family plays various roles in plant growth, plant development and flavonoid biosynthesis of secondary metabolism. In this study, the reported MYB transcription factors in F. tataricum were summarized and systemically clustered, and their interrelationships were defined to provide references for further exploring and cloning MYB transcription factors in F. tataricum. In addition, this study reviewed their regulatory functions of MYB transcription factors in flavonoid biosynthesis pathway, plant hormones pathways and other abiotic stress pathways, and made a conclusion and advances about the future research in F. tataricum. Therefore, this study will provide valuable scientific references for the functional studies of MYB family transcription factors in F. tataricum and its molecular breeding for high-quality varieties.