基于网络药理学和体内实验探讨天麻钩藤饮治疗内皮功能障碍的作用机制OA

Objective To investigate the mechanism of Tianma Gouteng Decoction(TGD)in ameliorating endothelial dysfunction through network pharmacology,molecular docking,and in vivo animal experiments.Methods(1)Network pharmacology research:Active components of TGD were screened from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and Swiss Target Prediction database.Disease targets related to endothelial dysfunction were obtained from the Online Mendelian Inheritance in Man(OMIM)and GeneCards databases.A"component-target-pathway"network was constructed.The STRING database was used to build a protein-protein interaction(PPI)network;Cytoscape software was employed to screen core targets;DAVID database was utilized for Gene Ontology(GO)analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.Molecular docking was performed to evaluate the binding affinity between key targets and major active components.(2)Animal experiments verification:An endothelial dysfunction model was induced in SD rats by intragastric administration of N ω-Nitro-L-arginine methyl ester hydrochloride(L-NAME)for 4 weeks,while the rats were concurrently treated with different doses of TGD and the positive drug nifedipine for 4 weeks.Systolic and diastolic blood pressure were measured.The mesenteric arterial vasodilation function was assessed using an isolated tension myograph.Serum levels of nitric oxide(NO),tumor necrosis factor-α(TNF-α),and interleukin-6(IL-6)were detected.Pathological morphology of the mesenteric arteries was examined by hematoxylin and eosin(HE)staining.Protein expression levels of the PI3K/AKT pathway and phosphorylated endothelial nitric oxide synthase(p-eNOS)in rat arteries were detected by Western Blot.Results A total of 198 active components and 1 081 potential targets of TGD were screened.Intersection with endothelial dysfunction disease targets yielded 425 core targets,which were primarily enriched in the PI3K-AKT signaling pathway.The main active components and their targets demonstrated favorable binding activity.Animal experiment results showed that TGD significantly reduced systolic and diastolic blood pressure in rats(P＜0.05,P＜0.01),enhanced acetylcholine-induced vasodilation(P＜0.01),significantly increased serum NO levels,and decreased IL-6 and TNF-α levels(P＜0.01).TGD also ameliorated mesenteric endothelial cell damage and significantly increased the ratios of phosphorylated phosphatidylinositol 3-kinase(p-PI3K)/PI3K,phosphorylated AKT serine/threonine kinase(p-AKT)/AKT,and p-eNOS/eNOS in rat arteries(P＜0.05,P＜0.01).Conclusion Tianma Gouteng Decoction can effectively treat endothelial dysfunction,and its mechanism may be related to the activation of the PI3K/AKT signaling pathway.