中药新药与临床药理2025，Vol.36Issue(2)：230-242,13.DOI:10.19378/j.issn.1003-9783.2025.02.009
基于网络药理学和实验验证探讨三七总皂苷调控自噬改善糖尿病肾病小鼠肾损伤的作用机制
Exploration on the Mechanism of Panax Notoginseng Saponins on Improving Renal Injury in Diabetic Nephropathy Mice by Regulating Autophagy Based on Network Pharmacology and Experimental Verification
摘要
Abstract
Objective To explore the mechanism of Panax notoginseng saponins(PNS)on protecting the kidney of diabetic nephropathy(DN)mice via regulating autophagy through network pharmacology,molecular docking and animal experiments.Methods The main active ingredients of PNS were obtained through literature search,and the corresponding targets were obtained using PubChem and PharmMapper databases.GeneCards and OMIM databases were used to collect DN related targets.After the intersection of active ingredients targets and disease targets was taken,Cytoscape 3.9.1 software was used to construct a"Chinese medicine-active ingredients-targets"network.A protein-protein interaction(PPI)network was constructed using STRING database and core targets were screened.The DAVID platform was applied to conduct Gene Ontology(GO)functional and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis on intersecting targets.AutoDock Vina software was used for molecular docking of the main active ingredients and key targets of PNS.Twenty-four 8-week-old male db/db mice were randomly divided into a model group,PNS group,and irbesartan group,with eight mice in each group.Another eight male db/m mice of the same age were selected as the normal group and orally administered once a day for 8 consecutive weeks.The general condition of mice in each group was observed and changes in body mass and blood glucose were recorded.Coomassie Brilliant Blue method was used to detect 24-hour urinary protein(24 h-UTP),the fructose amine method was applied to measure glycosylated serum protein(GSP),GPO-PAP method was used to measure total cholesterol(TC)and triglycerides(TG),creatine oxidase method was applied to measure serum creatinine(Scr),urease method was used to measure urea nitrogen(BUN).Renal pathological changes in mice were investigated using hematoxylin eosin(HE)staining.Real time fluorescence quantitative polymerase chain reaction(RT-qPCR)was used to detect the mRNA expression levels of ubiquitin binding protein(P62)and microtubule associated protein light chain 3(LC3)in the kidney tissues of mice in each group.The protein expressions of P62,LC3,phosphatidylinositol 3 kinase(PI3K),protein kinase B(Akt),mammalian target of rapamycin(mTOR)and phosphorylated proteins(p-Akt,p-PI3K,p-mTOR)in renal tissues were detected by Western Blot.Results The main active ingredients of PNS are notoginsenoside R1,ginsenoside Re,ginsenoside Rd,ginsenoside Rb1,and ginsenoside Rg1.Network pharmacology analysis showed that there were 341 targets corresponding to PNS active ingredients,1 534 targets corresponding to DN,and 113 intersecting targets between PNS and DN.The key components,which were screened from PNS in the treatment of DN,include ginsenosides Rd,Re,Rg1.The core targets are AKT1,ALB,MMP9,and mTOR.The related signaling pathways involve PI3K/Akt,Ras,MAPK,FoxO,etc.Molecular docking showed that the key components of PNS(ginsenosides Rd,Re,and Rg1)exhibited good docking activity with the core targets(AKT1,MMP9,and mTOR).The animal experiment results showed that compared with the normal group,the mice in model group had generally poorer state and significant increases in body weight,blood glucose,GSP,TC,TG,Scr,BUN,and 24 h-UTP(P<0.01).Compared with the model group,the mice in PNS treatment group showed an improvement in the general condition and significant decreases in body weight and blood glucose(P<0.01),as well as significant decreases in GSP,TC,TG,Scr,BUN,and 24 h-UTP(P<0.05,P<0.01).HE staining results showed a significant improvement in renal pathological damage of each treatment group.RT-qPCR and Western Blot results showed that compared with the normal group,the mRNA and protein levels of LC3 in the kidney tissues of mice in the model group significantly decreased(P<0.01),while the mRNA expression of P62 increased(P<0.01),and the protein expression of p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR and P62 also increased significantly(P<0.05,P<0.01).Compared with the model group,the mRNA and protein levels of LC3 in the PNS group and irbesartan group increased(P<0.05,P<0.01),the mRNA expression of P62 significantly decreased(P<0.05,P<0.01),and the protein expression of p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR and P62 significantly decreased(P<0.05,P<0.01).Conclusion PNS can significantly improve the renal damage in diabetic nephropathy mice,and its mechanism may be related to the inhibition of PI3K/Akt/mTOR signaling pathway and the promotion of autophagy.关键词
糖尿病肾病/三七总皂苷/自噬/网络药理学/分子对接/PI3K/Akt/mTOR/实验验证/小鼠Key words
Diabetic nephropathy/Panax notoginseng saponins/autophagy/network pharmacology/molecular docking/PI3K/Akt/mTOR/experimental verification/mice分类
医药卫生引用本文复制引用
陈勇,洪佳,朱宝璇,林榕榕,郑燕芳..基于网络药理学和实验验证探讨三七总皂苷调控自噬改善糖尿病肾病小鼠肾损伤的作用机制[J].中药新药与临床药理,2025,36(2):230-242,13.基金项目
国家自然科学基金项目(81973827) （81973827）
福建省自然科学基金项目(2021J01890) （2021J01890）
福建省大学生创新创业训练计划项目(202210393015,S202210393023). （202210393015,S202210393023）
