通络糖泰方治疗糖尿病周围神经病变靶点预测与机制探讨  

Target prediction and mechanism exploration of Tongluo Tangtai prescription in treatment of diabetic peripheral neuropathy

作　　者：陈世婷 梅英秀 陈明珠 杜联[1] CHEN Shi-ting;MEI Ying-xiu;CHEN Ming-zhu;DU Lian(Basic Medical College,Chengdu University of Traditional Chinese Medicine,Chengdu 611137,China)

机构地区：[1]成都中医药大学基础医学院,四川成都611137

出　　处：《中国药理学通报》2025年第4期772-780,共9页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No.81973585)。

摘　　要：目的基于网络药理学和体外实验验证探讨通络糖泰方(TLTT)防治糖尿病周围神经病变(DPN)的分子作用机制。方法通过TCMSP数据库及文献检索查找通络糖泰化学成分信息,运用PubChem、SwissTargetPrediction数据库收集化学成分作用靶点。通过GeneCards数据库收集DPN靶点;利用DAVID数据库对共有靶点蛋白进行GO功能及KEGG通路富集分析。使用Cytoscape软件构建相关网络图,筛选主要活性成分、靶点基因进行分子对接研究。高糖诱导雪旺细胞构建DPN体外模型,采用CCK-8法检测TLTT对RSC96细胞存活率影响,Real-time PCR检测RSC96细胞Wnt/β-catenin通路相关靶向分子的基因表达,Western blot检测RSC96细胞Wnt/β-catenin通路相关蛋白的表达水平。结果筛选豆甾醇、β-谷甾醇、槲皮素等主要活性成分,主要作用于EGFR、AKT1、MAPK3等关键靶点和Wnt、PI3K-Akt与MAPK等信号通路。分子对接结果显示,豆甾醇、β-谷甾醇、槲皮素等成分能与EGFR、AKT1、MAPK3等靶点良好对接。细胞实验结果显示,10%TLTT含药血清可促进高糖雪旺细胞活力,上调β-catenin蛋白表达,下调GSK-3β、Wif-1蛋白表达。结论TLTT治疗DPN具有多靶点-多通路的特性,其可能机制为TLTT激活Wnt/β-catenin信号通路,改善高糖对雪旺细胞的增殖抑制作用,促进雪旺细胞增殖,从而改善DPN的状态。Aim To explore the molecular mechanism of Tongluo Tangtai(TLTT)prescription in the prevention and treatment of diabetic peripheral neuropathy(DPN)based on network pharmacology and in vitro experimental verification.Methods The chemical composition information of Tongluo Tangtai was searched by TCMSP database and literature search,and the target of chemical composition was collected by PubChem and SwissTargetPrediction database.DPN targets were collected through GeneCards database.GO function and KEGG pathway enrichment of common target proteins were analyzed using DAVID database.The correlation network diagram was constructed using Cytoscape software,and the main active components and target genes were screened for molecular docking study.The in vitro model of DPN was constructed in Schwann cells induced by high glucose.The effect of TLTT on the survival rate of RSC96 cells was detected by CCK-8 method,and the gene expression of Wnt/β-catenin pathway related target molecules in RSC96 cells was detected by Real-time PCR.The expression levels of Wnt/β-catenin pathway-related proteins in RSC96 cells were detected by Western blot.Results The main active components such as stigmasterol,β-sitosterol and quercetin were screened,which mainly acted on EGFR,AKT1,MAPK3 and Wnt,PI3K-Akt and MAPK signaling pathways.The molecular docking results showed that stigmasterol,β-sitosterol,quercetin and other components could dock well with EGFR,AKT1,MAPK3 and other targets.The results of cell experiment showed that 10%TLTT drug-containing serum could promote the viability of high-glucose Schwann cells,up-regulate the expression ofβ-catenin protein,and down-regulate the expression of GSK-3βand Wif-1 protein.Conclusions TLTT has the characteristics of multi-target-multi-pathway in the treatment of DPN.The possible mechanism is that TLTT activates the Wnt/β-catenin signaling pathway,improves the inhibitory effect of high glucose on the proliferation of Schwann cells,promotes the proliferation of Schwann cells,and thus improves t

关 键 词：通络糖泰 糖尿病周围神经病变 网络药理学 分子对接 细胞实验 作用机制 

分 类 号：R-332[医药卫生] R285.5R319R329.28R587.2R745