基于网络药理学及实验验证探讨五子降糖方抑制骨骼肌IR的作用机制研究  

Study on the mechanism of wuzi Jiangtang formula Inhibiting skeletal muscle Insulin resistance based on network pharmacology and experimental verification

作　　者：张碧溦 王东军 徐丁洁 吴晨曦 韩茹杰 田朝阳 林浩 申争益 李继安 ZHANG Biwei;WANG Dongjun;XU Dingjie;WU Chenri;HAN Rujie;TIAN Zhaoyang;LIN Hao;SHEN Zhengyi;LI Ji'an(Traditional Chinese Medicine College,He Bei Key Laboratory of Integrated Traditional Chinese and Western Medicine for Diabetes and Its Com plications,North China University of Science and Technology,Tangshan O63000,China)

机构地区：[1]华北理工大学中医学院,河北省中西医结合防治糖尿病及其并发症重点实验室,河北省唐山市063210 [2]华北理工大学公共卫生学院

出　　处：《中国煤炭工业医学杂志》2024年第4期334-344,共11页Chinese Journal of Coal Industry Medicine

基　　金：河北省中医药管理局科研计划项目(编号:2020222,编号:2024355);河北省自然基金中医药联合基金重点项目(编号:H2023209038);河北省自然基金中医药联合基金(编号:H2023209049);2024年省级研究生专业学位教学案例库项目(编号:KCJSZ2024060);河北省自然科学基金(编号:H2020209123)。

摘　　要：目的 运用网络药理学与实验相结合的方法,初步探讨五子降糖方抑制骨骼肌胰岛素抵抗的作用机制。方法 从TCMSP、HERB、PubChem、PharmMapper数据库筛选出五子降糖方中5味中药的有效活性成分及其作用靶点;搜集并筛选骨骼肌胰岛素抵抗(IR)及细胞自噬相关治疗靶点;取三者交集,进行基因本体论(GO)和京都基因与基因组百科全书(KEGG)富集分析,并构建蛋白-蛋白相互作用(PPI)网络、成分-靶点-通路网络模型,筛选出核心靶点、关键成分及通路;运用AutoDuckTool、Vina、Pymol、Chem3D软件对药物关键成分及核心作用靶点进行分子对接。高脂高糖联合STZ构建糖尿病动物模型,通过ELISA法、HE染色以及Western blot检测验证网络药理学分析结果。结果 通过筛选分析共得到五子降糖方抑制骨骼肌IR和细胞自噬的作用靶点143个,GO和KEGG富集分析显示信号通路主要集中在磷脂酰肌醇-3-羟激酶-丝/苏氨酸蛋白激酶(PI3K-Akt)、肿瘤坏死因子(TNF)、糖基化终产物-晚期糖基化终末产物受体(AGERAGE)信号通路,涉及到的生物学过程主要有糖脂代谢、炎症反应、细胞自噬等。分子对接结果显示核心成分槲皮素、木犀草素、山奈酚、异鼠李素与关键靶点AKT1、肿瘤蛋白53(TP53)、TNF、白介素-6(IL-6)、PIK3CG对接良好。动物实验显示,与模型组比较,五子降糖方可降低胰岛素抵抗指数,升高PI3K、AKT蛋白表达及降低TNF-α蛋白表达。结论 本研究初步揭示了五子降糖方中的主要活性成分为槲皮素、木犀草素、山柰酚、异鼠李素等,其改善骨骼肌IR的作用可能与介导PI3K/Akt信号通路调控细胞自噬有关,为其基础研究及临床应用提供了依据。Objective To explore the mechanism of Wuzi Jiangtang Formula in inhibiting skeletal muscle insulin resistance(IR) through the integration of network pharmacology and experimental methods.Methods Active ingredients and their targets in Wuzi Jiangtang Formula were screened from TCMSP database, HERB database, PubChem database and PharmMapper database. Therapeutic targets related to skeletal muscle IR and autophagy were collected and filtered. GO and KEGG enrichment analysis was performed on the intersection of the three datasets, then both the PPI network and ingredient-target-pathway network model were con structed to identify the core targets, key ingredients, and pathways. Molecular docking was performed by softwares including AutoDockTool, Vina, Pymol, and Chem3D. A diabetic animal model was established by combining high-fat diet and streptozotocin(STZ), and the results of network pharmacological analysis were verified by ELISA, HE staining and Western blot.Results A total of 143 targets related to the inhibition of skeletal muscle IR and autophagy by Wuzi Jiangtang Formula were identified. GO and KEGG enrichment analyses revealed that the signaling pathways mainly focused on PI3K-Akt, TNF, and AGE-RAGE pathways, involving biological processes such as glucose and lipid metabolism, inflammation, and autophagy. Molecular docking results showed that all the core components such as quercetin, luteolin, kaempferol, and taxifolin were well docked with key targets including AKT1, TP53, TNF, IL6 and PIK3CG. Animal experiments demonstrated that compared with the model group, Wuzi Jiangtang Formula could reduce the insulin resistance index, increase PI3K and AKT protein expression, and decrease TNF-α protein expression. Conclusion This study preliminarily revealed that the main active ingredients in Wuzi Jiangtang Formula were quercetin, luteolin, kaempferol, and taxifolin, which may improve skeletal muscle IR by mediating the PI3K/Akt signaling pathway to regulate autophagy, providing a basis for its basic research and

关 键 词：五子降糖方 骨骼肌胰岛素抵抗 细胞自噬 网络药理学 分子机制 

分 类 号：R587.1[医药卫生—内分泌]