基于生物信息学和体外实验探讨金钗石斛治疗2型糖尿病的药理机制  

Pharmacological mechanism of Dendrobium nobile for type 2 diabetes mellitus by bioinformatics and in vitro experiment

作　　者：曾美玲 李朝阳 胡乔 耿克用 左振宇[2] 周威 ZENG Meiling;LI Zhaoyang;HU Qiao;GENG Keyong;ZUO Zhenyu;ZHOU Wei(School of Pharmacy,Guizhou Medical University,Guizhou Gui’an New District 561113,China;School of Pharmacy,Shaanxi University of Chinese Medicine,Shaanxi Xi’an 712046,China)

机构地区：[1]贵州医科大学药学院,贵州贵安新区561113 [2]陕西中医药大学药学院,陕西西安712046

出　　处：《中国医院药学杂志》2024年第14期1627-1634,共8页Chinese Journal of Hospital Pharmacy

基　　金：贵州省科技支撑计划项目(编号:黔科合支撑[2021]一般415);贵州省自然科学基金(编号:黔科合基础[2020]1Y404);贵州省大学生创新创业训练计划项目(编号:S202210660145,S202110660009)。

摘　　要：目的:通过生物信息学、体外实验探讨金钗石斛对2型糖尿病的治疗作用。方法:通过生物信息学数据库筛选出金钗石斛提取物治疗2型糖尿病的潜在靶点,构建蛋白相互作用(PPI)网络。通过DAVID数据库进行基因本体GO富集分析和KEGG通路富集分析,构建“药物-化合物-靶点-通路-疾病”网络。利用Autodock Vina 1.1.2对所筛选出的核心靶点与核心化合物进行分子对接。MTT法考察金钗石斛对链脲佐菌素(STZ)诱导的大鼠胰岛素瘤β(INS-1)细胞和小鼠胰岛素瘤β(NIT-1)细胞损伤的保护作用。Western blot方法考察金钗石斛提取物对NIT-1细胞的胰岛素信号通路核心靶点的影响,进行体外实验验证。结果:GO富集分析结果表明金钗石斛治疗2型糖尿病主要与胰岛素调节过程相关。KEGG通路富集分析结果表明金钗石斛通过PI3K-AKT信号通路、FOXO信号通路、糖尿病并发症中的AGE-RAGE信号通路、胰岛素抵抗、脂质与动脉粥样硬化以及胰岛素信号通路来治疗2型糖尿病。筛选出5个核心靶点,包括AKT1、SRC、EGFR、MAPK1、PIK3CA;6个核心化合物gigantol、moniliformin、moscatilin、tristin、dihydroconiferyl、trans-N-coumaroyltyramine。核心靶点与核心化合物具有良好的结合活性,结合能均不超过-6.7 kcal·mol^(-1)。金钗石斛提取物能够改善STZ诱导INS-1细胞和NIT-1细胞损伤,保护胰岛β细胞,发挥治疗2型糖尿病的作用。金钗石斛提取物能够提高受损胰岛β细胞的p-PI3K、p-PI3K/PI3K、p-AKT、pAKT/AKT的蛋白表达水平(P<0.01)。结论:金钗石斛提取物通过胰岛素信号通路中PI3K-AKT途径保护胰岛β细胞功能、改善胰岛素合成分泌,从而治疗临床2型糖尿病。OBJECTIVE To elucidate the pharmacological mechanism of Dendrobium nobile for type 2 diabetes mellitus(T2DM)by bioinformatics and in vitro experiment.METHODS Potential targets of D.nobile extract for treating T2DM were screened through bioinformatic databases for constructing protein-protein interaction(PPI)network.GO enrichment analysis of gene ontology and KEGG pathway enrichment analysis were performed with DAVID database for constructing a“DrugCompound-Target-Pathway-Disease”network.Autodock Vina 1.1.2 was utilized for molecular docking of selected core targets and core compounds.MTT method was utilized for examining the protective effect of Dendrobium nobile on streptozotocin(STZ)-induced insulinomaβ(INS-1)cell injury model and insulinomaβ(NIT-1)cell injury model.The effect of D.nobile extract on core targets of insulin signaling pathway in NIT-1 cells was analyzed by Western blot and in vitro experiment.RESULTS The results of GO enrichment analysis showed that the treatment of T2DM by Dendrobium nobile was mainly correlated with the process of insulin regulation.The results of KEGG pathway enrichment analysis revealed that Dendrobium nobile treated T2DM through the signaling pathways of PI3K-AKT,FOXO and AGE-RAGE in diabetic complications,insulin resistance,lipid and atherosclerosis and insulin signaling pathway.Five core targets of AKT1,SRC,EGFR,MAPK1 and PIK3CA were selected.Six core compounds were gigantol,moniliformin,moscatilin,tristin,dihydroconiferyl and trans-Ncoumaroyltyramine.Core targets had decent binding activities with core compounds and binding energy was<-6.7 kcal·mol^(-1).Dendrobium nobile extract could improve the injury of INS-1/NIT-1 cells induced by STZ,protect isletβcells and play a role in treating T2DM.Dendrobium nobile extract could boost the protein expression levels of p-PI3K,p-PI3K/PI3K,p-AKT and pAKT/AKT in injured pancreaticβcells(P<0.01).CONCLUSION Dendrobium nobile extract protects the function of pancreatic isletβcells and improves insulin synthesis and secretion t

关 键 词：金钗石斛 2型糖尿病 生物信息学 体外实验 

分 类 号：R285[医药卫生—中药学] R969[医药卫生—中医学]