基于网络药理学探究黄芪-黄连药对治疗2型糖尿病的机制

Mechanism of Astragali Radix-Coptis Rhizoma pair in treating type 2 diabetes mellitus based on network pharmacology

利用网络药理学和动物实验方法探讨黄芪-黄连药对治疗2型糖尿病(T2DM)的主要机制。通过CNKI数据库检索出治疗糖尿病的主要中药复方并找出使用频率最高的中药。通过TCMSP、TCMID数据库及文献检索中药的有效成分。利用SwissTargetPrediction和SEA数据库查找成分的作用靶点。利用Disgenet、TTD和DrugBank查找T2DM相关的疾病靶点。利用Cytoscape软件构建中药-成分-疾病-靶点网络图并找到黄芪-黄连药对治疗T2DM的作用靶点。通过STRING数据库查找作用靶点的相关蛋白并通过Cytoscape软件构建PPI网络图。利用ClueGO软件对PPI网络进行生物学功能分析。最后采用高脂高糖饲料喂养结合链脲佐菌素(STZ)30 mg·kg^(-1)一次性腹腔注射复制T2DM大鼠模型,分为对照组,模型组,阳性药二甲双胍组和黄芪-黄连药对组,给药时间持续1个月。生化法检测各组用药后血糖、血脂的变化。HE染色观察各组胰腺和胰岛结构,Masson染色观察胰腺和胰岛组织胶原沉积的变化。结果发现黄芪-黄连可用作治疗T2DM的中药药对。Ras相关C3肉毒毒素底物1(ras-related C3 botulinum toxin substrate 1,RAC1)、对氧磷脂酶1(paraoxonase 1,PON1)、β-半乳糖苷α2,6-唾液酸转移酶(α2,6-sialyltransferase,ST6GAL1)、胰岛素受体(insulin receptor,INSR)、性激素结合球蛋白(sex hormone binding globulin,SHBG)、钠依赖性胆汁酸转运体(sodium-dependent bile acid transporter,SLC10A2)、内皮素受体A(endothelin receptor A,EDNRA)、过氧化物酶体增殖物激活受体A(peroxisome proliferator-activated receptor A,PPARA)、内皮素受体B(endothelin receptor B,EDNRB)和羟色胺受体2A(hydroxytryptamine receptor 2A,HTR2A)是黄芪和黄连药对治疗T2DM的作用靶点。黄芪-黄连药对治疗T2DM的主要生物学功能是调节血糖和血脂代谢及对心血管系统的保护作用。模型组7、14、28 d空腹血糖值及甘油三酯(triglyceride,TG)、胆固醇(cholesterol,CHO)、低密度脂蛋白(low density lipoprotein,LDL)水平较对照组均显著升高(P<0.01),高密度脂蛋白(HDL,high density lipoprotein)含量显著降低(P<0.05)。二甲双胍组及黄芪-黄连药对组给药14、28 d大鼠的空腹血糖值较模型组明显降低(P<0.05),血清中TG、CHO、LDL含量较模型组显著降低(P<0.05),HDL含量显著增加(P<0.05)。黄芪-黄连药与二甲双胍组血糖、TG、CHO、LDL、HDL含量差异均无统计学意义。模型组胰腺和胰岛组织结构明显破坏,胰腺和胰岛组织胶原沉积明显增多且细胞边界不清。二甲双胍和黄芪-黄连药对组胰腺和胰岛结构的破坏和胶原化程度明显减轻。黄芪-黄连药对能有效调节T2DM的糖脂代谢紊乱,并通过调节心血管系统保护胰腺和胰岛的结构和细胞功能,值得中医临床和新药研发推广运用。

This study aims to explore the main mechanism of Astragali Radix-Coptis Rhizoma pair(hereinafter referred to as the pair)in the treatment of type 2 diabetes mellitus(T2DM)based on network pharmacology and animal experiment.The main Chinese medicine compound prescriptions for T2DM were retrieved from CNKI database and the medicinals with high frequency among these prescriptions were screened.The active components in the above medicinals were searched from TCMSP,TCMID,and previous research,targets of the components from SwissTargetPrediction and SEA,and targets for the treatment of T2DM from DISGENET,TTD,and DrugBank.Thereby,the medicinal-component-disease-target network was constructed with Cytoscape.The targets were input in String database to yield the related proteins and the protein-protein interaction(PPI)network was constructed by Cytoscape.The biological functions of proteins in the PPI network were analyzed by Cluego.Then,high-fat high-sugar diet and 30 mg·kg^(-1) streptozotocin(STZ,intraperitoneal injection,once)were employed to induce T2DM in rats and the T2DM rats were classified into the control group,model group,positive drug(metformin)group,and pair group.After one month of administration,the changes of blood glucose and blood lipids[triglyceride(TG),cholesterol(CHO),low density lipoprotein(LDL),high density lipoprotein(HDL)]were detected with biochemical methods and pathological changes of islet and collagen deposition in pancreatic tissue by HE staining and Masson staining,respectively.The result showed that pair can be used for T2DM treatment.ras-related C3 botulinum toxin substrate 1(RAC1),paraoxonase 1(PON1),beta-galactoside alpha 2,6-sialyltransferase 1(ST6 GAL1),insulin receptor(INSR),sex hormone-binding globulin(SHBG),ileal sodium/bile acid cotransporter(SLC10 A2),endothelin-1 receptor A(EDNRA),peroxisome proliferator-activated receptor A(PPARA),endothelin receptor B(EDNRB),and 5-hydroxytryptamine receptor 2A(HTR2A)were the targets of the pair for the treatment of T2DM.The main biological functions of the pair were regulating the metabolism of blood glucose and li-pids and protecting the cardiovascular system.The fasting blood glucose,and serum TG,CHO,and LDL were higher(P<0.01)and the HDL was lower(P<0.05)in the model group than in the control group on the 7 th,14 th,and 28 th days.The fas-ting blood glucose and the serum TG,CHO,and LDL decreased(P<0.05)and the serum HDL increased(P<0.05)in the metformin group and the pair group as compared with those in the model group on the 14 th and 28 th days.There were no significant differences in blood glucose,TG,CHO,LDL,and HDL between the metformin group and the pair group.Rats in the model group demonstrated damaged structures of islets and pancreas,obviously increased deposition of collagen in islets and pancreas,and blurred cell boundaries.Metformin and the pair significantly alleviated the damaged structures and collagen deposition.The pair can effectively regulate the disorders of blood glucose and lipid metabolism in T2DM and protect the structure and functions of pancreas and islets by controlling cardiovascular system,which is worthy of clinical application and can be used for drug development.