补阳还五汤对糖尿病引起的代谢相关脂肪性肝病治疗作用的网络药理学与实验研究

Therapeutic Effect of Buyang Huanwu Decoction(补阳还五汤)on Metabolic Associated Fatty Liver Disease Caused by Diabetes Mellitus:A Study Based on Network Pharmacology and Animal Experiment

目的:通过网络药理学探究补阳还五汤对糖尿病引起的代谢相关脂肪性肝病的治疗作用,并通过动物实验进行验证。方法:通过TCMSP数据库收集药物成分并进行靶点预测,从Disgenet数据库、Drugbank数据库、HPO数据库获取疾病相关靶点,David 6.8数据库进行共有靶点GO生物学过程分析和KEGG通路分析,String数据库获得关键靶点;建立2型糖尿病模型,设置模型对照组、二甲双胍0.2 g/kg组和补阳还五汤12.6 g/kg组,另设正常对照组,给药相应药物6 w,检测动物血糖(FBG)、血清三酰甘油(TG)、胆固醇(TC)的含量、天冬氨酸氨基转移酶(AST)和丙氨酸氨基转移酶(ALT)的活力,计算肝脏的脏器指数,HE观察大鼠肝脏病理变化,Western Blot检测肝脏的相关蛋白表达。结果:网络药理学分析显示,补阳还五汤与疾病共有靶点68个,生物学过程与调节信号通路的激酶、细胞凋亡、炎性因子、物质代谢有关,其作用通路与PPAR信号通路、PI3K-AKT信号通路、HIF-1信号通路等有关,关键靶点为AKT1、TNF、PPARG、VEGFA等。动物实验显示,与模型对照组相比,补阳还五汤12.6 g/kg组能明显降低糖尿病大鼠的血清FBG、TC、TG、ALT和AST的含量或活力,使肝脏组织的脂肪病变情况减轻,上调肝组织中PI3K、AKT、Beclin-1蛋白表达,下调Caspase-3、NF-κB、AGER蛋白表达(P<0.05或P<0.01)。结论:补阳还五汤可以调节PI3K-AKT信号通路,调节细胞的自噬、凋亡和炎症反应,对糖尿病引起的代谢相关脂肪性肝病产生治疗作用。

Objective:To reveal the effect of Buyang Huanwu Decoction(补阳还五汤,BHD)on metabolic associated fatty liver disease(MAFLD)caused by diabetes based on network pharmacology and verify the results through animal experiments.Methods:The active components of the decoction were screened out from TCMSP,and then their targets were predicted.The targets of MAFLD were obtained from Digenet,Drugbank,and HPO.David 6.8 was used to perform the KEGG pathway enrichment and GO annotation of the common targets shared by active components and MAFLD,and the key targets were determined with STRING.TypeⅡdiabetes rat model was established,and then the rats were assigned into normal(Nor)group,model(Mod)group,metformin 0.2 g/kg(Met)group,and Buyang Huanwu Decoction 12.6 g/kg(BHD)group.All the rats were administrated for 6 weeks.The levels of fasting blood glucose(FBG),total cholesterol(TC),triglyceride(TG),alanine aminotransferase(ALT),and aspartate aminotransferase(AST)were detected.The liver organ index was calculated,and the pathological changes of rat liver were observed.Finally,western blotting was employed to determine the expression levels of liver-related proteins.Results:A total of 68 common targets shared by BHD active components and MAFLD were screened out,which were related to kinase,apoptosis,inflammatory cytokines,and material metabolism.The enriched pathways mainly included PPAR,PI3 K-Akt,and HIF-1 signaling pathways,and the key targets were AKT1,TNF,PPARG,VEGFA,etc.The animal experiments showed that compared with the model group,12.6 g/kg BHD reduced the levels of FBG,TG,TC,ALT,and AST in diabetic rats,alleviated the pathological changes in liver fatty tissue,up-regulated the protein levels of PI3 K,Akt,and Beclin-1,and down-regulated the protein levels of Caspase-3,NF-κB,and AGER.Conclusion:BHD can regulate PI3 K-Akt signaling pathway,autophagy,apoptosis,and inflammatory reaction to exert the therapeutic effect on MAFLD caused by diabetes.