基于网络药理学、分子对接和动物实验探究黄芪桂枝五物汤预防运动性疲劳的作用  

Effects of Huangqi Guizhi Wuwu(黄芪桂枝五物)Decoction in the Treatment of Exercise-Induced Fatigue Based on Network Pharmacology,Molecular Docking and Animal Experiments

作　　者：冉清智 何汶洋 李傲霜 陈恒文[1] 吴淮[2] 何本祥 RAN Qingzhi;HE Wenyang;LI Aoshuang;CHEN Hengwen;WU Huai;HE Benxiang(Guang'anmen Hospital,China Academy of Traditional Chinese Medicine,Beijing 100053;The Fifth Clinical Medical College of Guangzhou University of Chinese Medicine(The Second Chinese Medicine Hospital of Guangdong Province),Guangzhou 510095;Dongzhimen Hospital,Beijing University of Chinese Medicine,Beijing 100070;Sichuan Academy of Traditional Chinese Medicine,Chengdu 610042)

机构地区：[1]中国中医科学院广安门医院,北京100053 [2]广州中医药大学第五临床医学院(广东省第二中医院),广州510095 [3]北京中医药大学东直门医院,北京100070 [4]四川省中医药科学院,成都610042

出　　处：《中药药理与临床》2024年第6期18-25,共8页Pharmacology and Clinics of Chinese Materia Medica

基　　金：中央高水平中医医院临床科研业务费(编号:HLCMHPP2023077);中国中医科学院广安门医院续航人才工程青年拔尖人才培养项目(编号:CZ40909);中国中医科学院科技创新工程重大攻关项目(编号:CI2021A05011);中国中医科学院科技创新工程创新团队项目(编号:CI2021B017-05);国家自然科学基金(编号:82074396);国家重点研发计划项目(编号:2019YFF0301704)。

摘　　要：目的:采用网络药理学-分子对接的方法,探讨黄芪桂枝五物汤预防运动性疲劳(Exercise-induced fatigue,EF)的作用机制,并进行动物实验验证分析。方法:通过网络药理学工具获得黄芪桂枝五物汤预防EF的有效成分、核心靶点和信号通路。构建EF大鼠模型,进行HE染色、ELISA、PCR法验证网络药理学预测的主要靶点及预防效果。结果:筛选出黄芪桂枝五物汤中43个活性成分和242个潜在靶点,运动性疲劳潜在靶点1002个;药物和疾病交集靶点104个,包括AKT1、IL6、TNF、IL1B、PPARG、PTGS2等。KEGG富集分析结果包括AGE-RAGE信号通路、TNF信号通路、IL-17信号通路、HIF-1信号通路等;分子对接结果显示豆甾醇、刺芒柄花素及原阿片碱等活性成分与PTGS2、PTGS1、CHRM1等关键靶点表现出良好的对接活性,黄芪桂枝五物汤可能通过影响AKT1、IL-6、TNF、IL-1B、PPARG、PTGS2等靶蛋白的表达进而调控炎症反应和细胞凋亡。黄芪桂枝五物汤可以使大鼠肌纤维排列更紧密,增强大鼠四肢肌力、运动耐力、心脏供血能力,且缓解心脏过度代偿。与正常对照组比较,模型对照组游泳力竭时间与四肢抓力显著减少,血清中肌酸激酶(CK)、血乳酸(BLA)、白介素6(IL-6)、肿瘤坏死因子(TNF-α)、缺氧诱导因子(HIF-1α)的含量显著升高,血管内皮生长因子(VEGF)与促红细胞生成素(EPO)的含量显著降低(P<0.01),雌激素受体1(Esr1)表达下调,前列腺素内过氧化物合酶2(Ptgs2)、蛋白激酶1(Akt1)、过氧化物酶体增殖物激活受体γ(Pparg)和果寡糖(Fos)mRNA表达上调(P<0.05);与模型对照组比较,黄芪桂枝五物汤组游泳力竭时间与四肢抓力显著增加,血清中CK、BLA、IL-6、TNF-α、HIF-1α的含量显著减少;VEGF、EPO的含量显著增加(P<0.01);Esr1 mRNA表达上调,Ptgs2、Akt1、Pparg、Fos mRNA表达下调(P<0.05)。结论:黄芪桂枝五物汤中槲皮素、β-谷甾醇、山柰酚等化合物发挥核心作用,�Objective:To investigate the mechanism of action of Huangqi Guizhi Wuwu(黄芪桂枝五物)Decoction in the treatment of exercise-induced fatigue(EF)by using network pharmacology and molecular docking,and to validate and analyze it through animal experiments.Methods:The active ingredients,core targets and signaling pathways of Huangqi Guizhi Wuwu Decoction in the treatment of EF were obtained by network pharmacology tools.An EF rat model was constructed,and HE staining,ELISA,and PCR experiments were performed to verify the principal targets and therapeutic effects predicted by network pharmacology.Results:A total of 43 active ingredients and 242 potential targets in Huangqi Guizhi Wuwu Decoction were screened out,and 1002 potential targets for EF.A total of 104 targets was observed at the intersection of drug and disease,including AKT1,IL6,TNF,IL1B,PPARG,PTGS2,etc.The results of KEGG enrichment analysis showed that AGERAGE,TNF,IL-17,TNF,IL-17and HIF-1 signaling pathways were involved.The molecular docking results showed that the active ingredients,such as leguminol,prickly aristolochicin and proto-opiomelanocarpine hadgood binding activities with key targets,such as PTGS2,PTGS1,CHRM1,etc.Huangqi Guizhi Wuwu Decoction affected the expression of target proteins,such as AKT1,IL-6,TNF,IL-1B,PPARG,PTGS2,etc.,and thus regulated inflammatory response and apoptosis.Animal experiment results showed that Huangqi Guizhi Wuwu Decoction could induce rat muscle fibers more closely aligned,enhance rat limb muscle strength,exercise endurance,cardiac blood supply,and alleviate the cardiac overcompensation.Compared with the control group,the swimming exhaustion time and limb grip strength were significantly reduced in the model group.The serum levels of creatine kinase(CK),blood lactate(BLA),interleukin 6(IL-6),tumor necrosis factor-α(TNF-α),and hypoxia-inducible factor-1α(HIF-1α)were significantly elevated.The levels of vascular endothelial growth factor(VEGF)and erythropoietin(EPO)were significantly reduced(P<0.01).The mRNA

关 键 词：黄芪桂枝五物汤 运动性疲劳 网络药理学 分子对接 过氧化物酶体增殖物激活受体Γ 果寡糖 前列腺素内过氧化物合酶2 

分 类 号：R285.5[医药卫生—中药学]