目的 应用网络药理学的研究方法筛选人参—黄连—三七药串治疗糖尿病肾病的作用靶点及相关信号通路,通过动物实验明确其疗效与作用机制。方法 运用Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(T...目的 应用网络药理学的研究方法筛选人参—黄连—三七药串治疗糖尿病肾病的作用靶点及相关信号通路,通过动物实验明确其疗效与作用机制。方法 运用Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)数据库检索获取人参—黄连—三七的主要化学成分以及相关作用靶点;使用DisGeNET筛选胰岛素抵抗与糖尿病肾病的相关靶标基因;通过Venn软件筛选出药物与疾病的共同作用靶点;构建出疾病—靶点—成分—药物网络;使用STRING数据库完成蛋白质—蛋白质相互作用网络(protein-protein interaction networks, PPI)的构建;通过基因本体(gene ontology, GO)富集分析和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes, KEGG)通路富集分析对有效作用靶点进行分析。以db/db小鼠作为动物模型,于给药第4、第8周后留取各组小鼠8小时尿液,检测各组小鼠尿微量白蛋白排泄水平;采用蛋白免疫印迹法法对网络药理学的研究预测的关键通路及靶点进行进一步的实验验证。结果 本研究共筛选得到人参—黄连—三七药串的人参皂苷Rh2、人参皂苷F2、槲皮素、小檗碱等39种有效成分;获得人参—黄连—三七药串治疗糖尿病肾病潜在靶点18个,其中PTEN、CCL2、IL6、TNF、HIF1A等11个与其他靶点相互作用较强,可能在网络中起到关键作用;通过GO分析和KEGG分析分别获得2595个条目和107条信号通路,其中包括磷脂酰肌醇-3-激酶丝氨酸/苏氨酸特异性蛋白激酶(phosphatidylinositol-3-kinase/protein kinase B,PI3K/Akt)信号通路、晚期糖基化终末产物—晚期糖基化终末产物受体信号通路、低氧诱导因子-1信号通路、细胞迁移的正调控、细胞因子活性、细胞因子受体结合、血小板α颗粒等。动物实验显示,在给药第4周及第8周后,中药组小鼠尿微量白蛋白排泄水平均较模型组下降(P<0.05)。对于PI3K/Akt信号通路的相关蛋白检测结果表明,中药组小鼠肾脏PI3K、p-Akt表达水平较模型组明显下降(P<0.05),磷酸酶张力蛋白同源物(phosphatase and tensin homolog deleted on chromosome ten, PTEN)水平较模型组升高(P<0.05)。结论 人参—黄连—三七药串能够有效改善db/db小鼠糖尿病肾病引起的蛋白尿,其作用机制可能与激活PTEN抑制PI3K/Akt信号通路有关。展开更多
目的筛选人参、黄连药对主要活性成分,预测其活性成分的作用靶点,建立药物—活性成分—作用靶点网络,探讨人参、黄连药对治疗2型糖尿病(diabetes mellitus type 2,T2DM)的作用机制。方法使用TCMID、TCMSP平台筛选人参、黄连药对化学成分...目的筛选人参、黄连药对主要活性成分,预测其活性成分的作用靶点,建立药物—活性成分—作用靶点网络,探讨人参、黄连药对治疗2型糖尿病(diabetes mellitus type 2,T2DM)的作用机制。方法使用TCMID、TCMSP平台筛选人参、黄连药对化学成分,并应用STITCH及DrugBank数据库检索活性成分靶点。应用TTD、DrugBank、GAD和DisGeNET数据库,筛选T2DM相关靶点。采用Cytoscape软件构建药对活性成分—作用靶点网络,通过David数据库对靶点进行生物学功能及通路富集分析。结果筛选得到人参、黄连药对潜在活性成分46个,人参相关靶点144个,黄连相关靶点246个,收集到350个T2DM相关靶点。相匹配后,得到42个人参、黄连药对治疗T2DM潜在靶点,其中,槲皮素(quercetin)、黄连素(berberine)、山奈酚(kaempferol)、花生四烯酸(arachidonate acid)、阿魏酸(ferulic acid)、豆甾醇(Stigmasterol)等6个成分能与3个及以上的关键靶点基因相连,可能是人参、黄连中治疗2型糖尿病的重要成分。结论人参、黄连药对靶点和药物代谢、胰岛素抵抗、炎症反应相关,并且参与了AMPK信号通路、NF-κB信号通路、TNF信号通路以及胰高血糖素信号通路,这些生物过程和信号通路可能是人参、黄连药对治疗T2DM的潜在机制。展开更多
OBJECTIVES:To observe the efficacy of Shenlian formula(SL formula,参连方),which consist of Huanglian(Rhizoma Coptidis)and Renshen(Radix Ginseng),in the treatment of type 2 diabetes mellitus(T2DM)and explore the effect...OBJECTIVES:To observe the efficacy of Shenlian formula(SL formula,参连方),which consist of Huanglian(Rhizoma Coptidis)and Renshen(Radix Ginseng),in the treatment of type 2 diabetes mellitus(T2DM)and explore the effects on gut microbiota and serum inflammatory cytokines.METHODS:In a double-blind,randomized,placebocontrolled parallel-group clinical trial,31 adults with T2DM were randomly allocated to receive the SL formula or placebo for 12 weeks.Body mass index(BMI),blood lipid indices,glycemic biomarkers including glycated hemoglobin(HbA1C),fasting plasma glucose(FPG),postprandial blood glucose(PBG),fasting insulin levels(FIL),fasting C-peptide(C-P),homoeostasis model assessment for insulin resistance(HOMA-IR)and inflammatory cytokines were assessed at baseline and 12 weeks.The contents of gut microbiota were determined by pyrosequencing of the V3-V4 regions of 16S rRNA genes.RESULTS:Sixteen cases were allocated in the treatment group and 15 in the placebo group.Compared with the placebo,SL formula resulted in a higher significant reduction in PBG[(-1.318±0.772)vs(-0.008±1.404)mmol/L,P=0.003],BMI[(-0.611±0.524)vs(0.957±2.212)kg/m2,P=0.01],FIL[(-1.627±6.268)vs(3.976±6.85)μIU/mL,P=0.02],HOMA-IR[(-0.530±1.461)vs(1.511±2.288),P=0.006],and C-reactive protein(CRP)[(-1.307±0.684)vs(0.828±0.557)mg/L,P=0.04].In terms of gut microbiota,compared with the placebo,the SL formula resulted in a significant decrease in species richness and evenness.CONCLUSIONS:The SL formula showed the efficacy to improve postprandial blood glucose,insulin resistance,BMI and CRP levels.In addition,it could reduce the total number,richness and evenness of species,meanwhile increase the abundance of probiotics to modulate the structure of gut microbiota in patients with T2DM.However,further studies are required for exploring the deeper mechanism of TCM on gut microbiota.展开更多
文摘目的 应用网络药理学的研究方法筛选人参—黄连—三七药串治疗糖尿病肾病的作用靶点及相关信号通路,通过动物实验明确其疗效与作用机制。方法 运用Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)数据库检索获取人参—黄连—三七的主要化学成分以及相关作用靶点;使用DisGeNET筛选胰岛素抵抗与糖尿病肾病的相关靶标基因;通过Venn软件筛选出药物与疾病的共同作用靶点;构建出疾病—靶点—成分—药物网络;使用STRING数据库完成蛋白质—蛋白质相互作用网络(protein-protein interaction networks, PPI)的构建;通过基因本体(gene ontology, GO)富集分析和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes, KEGG)通路富集分析对有效作用靶点进行分析。以db/db小鼠作为动物模型,于给药第4、第8周后留取各组小鼠8小时尿液,检测各组小鼠尿微量白蛋白排泄水平;采用蛋白免疫印迹法法对网络药理学的研究预测的关键通路及靶点进行进一步的实验验证。结果 本研究共筛选得到人参—黄连—三七药串的人参皂苷Rh2、人参皂苷F2、槲皮素、小檗碱等39种有效成分;获得人参—黄连—三七药串治疗糖尿病肾病潜在靶点18个,其中PTEN、CCL2、IL6、TNF、HIF1A等11个与其他靶点相互作用较强,可能在网络中起到关键作用;通过GO分析和KEGG分析分别获得2595个条目和107条信号通路,其中包括磷脂酰肌醇-3-激酶丝氨酸/苏氨酸特异性蛋白激酶(phosphatidylinositol-3-kinase/protein kinase B,PI3K/Akt)信号通路、晚期糖基化终末产物—晚期糖基化终末产物受体信号通路、低氧诱导因子-1信号通路、细胞迁移的正调控、细胞因子活性、细胞因子受体结合、血小板α颗粒等。动物实验显示,在给药第4周及第8周后,中药组小鼠尿微量白蛋白排泄水平均较模型组下降(P<0.05)。对于PI3K/Akt信号通路的相关蛋白检测结果表明,中药组小鼠肾脏PI3K、p-Akt表达水平较模型组明显下降(P<0.05),磷酸酶张力蛋白同源物(phosphatase and tensin homolog deleted on chromosome ten, PTEN)水平较模型组升高(P<0.05)。结论 人参—黄连—三七药串能够有效改善db/db小鼠糖尿病肾病引起的蛋白尿,其作用机制可能与激活PTEN抑制PI3K/Akt信号通路有关。
基金the National Key R&D Program of China:the Study on Syndrome Differentiation Standard of Yin Deficiency Syndrome in Type 2 Diabetes Mellitus(2018YFC1704402)the Project for Young Teachers of Beijing University of Chinese Medicine:Clinical Study on Intervention of Shenlian Formula based on Microbiota in Type 2 Diabetes Mellitus(No.2018-JYBZZ-JS062)。
文摘OBJECTIVES:To observe the efficacy of Shenlian formula(SL formula,参连方),which consist of Huanglian(Rhizoma Coptidis)and Renshen(Radix Ginseng),in the treatment of type 2 diabetes mellitus(T2DM)and explore the effects on gut microbiota and serum inflammatory cytokines.METHODS:In a double-blind,randomized,placebocontrolled parallel-group clinical trial,31 adults with T2DM were randomly allocated to receive the SL formula or placebo for 12 weeks.Body mass index(BMI),blood lipid indices,glycemic biomarkers including glycated hemoglobin(HbA1C),fasting plasma glucose(FPG),postprandial blood glucose(PBG),fasting insulin levels(FIL),fasting C-peptide(C-P),homoeostasis model assessment for insulin resistance(HOMA-IR)and inflammatory cytokines were assessed at baseline and 12 weeks.The contents of gut microbiota were determined by pyrosequencing of the V3-V4 regions of 16S rRNA genes.RESULTS:Sixteen cases were allocated in the treatment group and 15 in the placebo group.Compared with the placebo,SL formula resulted in a higher significant reduction in PBG[(-1.318±0.772)vs(-0.008±1.404)mmol/L,P=0.003],BMI[(-0.611±0.524)vs(0.957±2.212)kg/m2,P=0.01],FIL[(-1.627±6.268)vs(3.976±6.85)μIU/mL,P=0.02],HOMA-IR[(-0.530±1.461)vs(1.511±2.288),P=0.006],and C-reactive protein(CRP)[(-1.307±0.684)vs(0.828±0.557)mg/L,P=0.04].In terms of gut microbiota,compared with the placebo,the SL formula resulted in a significant decrease in species richness and evenness.CONCLUSIONS:The SL formula showed the efficacy to improve postprandial blood glucose,insulin resistance,BMI and CRP levels.In addition,it could reduce the total number,richness and evenness of species,meanwhile increase the abundance of probiotics to modulate the structure of gut microbiota in patients with T2DM.However,further studies are required for exploring the deeper mechanism of TCM on gut microbiota.
