基于网络药理学、分子对接和实验验证研究续断散加当归、骨碎补调控PI3K/AKT信号通路治疗激素性股骨头坏死的潜在机制  

Study on Potential Mechanism of Teasel Root Sanjia Angelica Sinensis and Drynaria Rhizome Regulating PI3K/AKT Signal Pathway to Treat Steroid-Induced Femoral Head Necrosis Based on Network Pharmacology,Molecular Docking,and Experimental Verification

作　　者：赵灿斌 付仔祥 王凤铭 郭超 李晓阳 ZHAO Can-bin;FU Zi-xiang;WANG Feng-ming;GUO Chao;LI Xiao-yang(Shandong University of Traditional Chinese Medicine,Jinan Shandong 250014;Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan Shandong 250014)

机构地区：[1]山东中医药大学,山东济南250014 [2]山东中医药大学附属医院,山东济南250014

出　　处：《世界中西医结合杂志》2023年第7期1366-1374,共9页World Journal of Integrated Traditional and Western Medicine

基　　金：山东省自然科学基金(ZR2022MH147);山东省中医药科技发展项目(2020Q016)。

摘　　要：目的基于网络药理学、分子对接技术,结合动物实验验证探究续断散加当归、骨碎补(Teasel Root Sanjia Angelica Sinensis and Drynaria Rhizome,TASDR)对激素性股骨头坏死的治疗作用,并初步探讨其潜在作用机制。方法采用TCMSP数据库检索TASDR中各味中药的化学成分和中药靶点,利用GeneCards数据库筛选激素性股骨头坏死的作用靶点,通过String平台构建核心靶点基因的蛋白质PPI(蛋白质互相作用)网络,通过Omic-Share Tools进行GO、KEGG分析。使用AutoDock 1.5.6软件对所得靶点以及姜黄素进行分子对接验证。动物实验选取24只新西兰大白兔,分为空白组,模型组及高、低剂量药物干预组,从中选取18只建立醋酸泼尼松龙诱导的激素性股骨头坏死模型,各组采取相应的药物干预,采用HE染色、免疫组化、Western blot检测,比较TASDR在治疗激素性股骨头坏死过程中对PI3K/AKT信号通路中某些关键蛋白表达水平的影响。结果经过筛选后TASDR治疗激素性股骨头坏死潜在核心靶点38个,靶点包括TNF、IL-6、VEGFA等,此外TASDR作用靶点中包括AKT蛋白家族-AKT1。TASDR通过肿瘤信号通路、VEGF信号通路等通路,并涉及酶结合、受体结合等分子功能,参与程序性细胞凋亡、程序性细胞凋亡负调控等生物过程来调控成骨分化。分子对接显示TASDR与TNF、VEGF、AKT1等靶点基因的结合活性良好。动物实验结果表明:在模型组中,PI3K、AKT等蛋白呈现高表达,而下游蛋白FOXO1及VEGF表达量减少,经TASDR干预后PI3K及AKT等蛋白表达量下降,FOXO1及VEGF等蛋白表达升高。结论TASDR有着靶点、通路多元化的优势,可在一定程度上调控TNF、IL-6、VEGF、AKT1等靶点基因的表达,动物实验表明PI3K/AKT信号通路过度激活时,TASDR可一定程度上下调上游蛋白AKT及PI3K的表达量,且使得下游基因FOXO1及VEGF的表达量增加进而在激素性股骨头坏死的治疗中发挥作用。Objective Based on network pharmacology,molecular docking technology,and animal experimental verification,the therapeutic effect of Teasel Root Sanjia Angelica Sinensis and Drynaria Rhizome(TASDR)on steroid-induced femoral head necrosis was explored,and its potential mechanism was preliminarily discussed.Methods TCMSP database was used to search for the chemical components and targets of various traditional Chinese medicines in TASDR,and the GeneCards database was used to screen the targets of steroid-induced femoral head necrosis.A protein-protein interaction(PPI)network of core target genes was constructed through the String platform,and OmicShare Tools was used to analyze GO and KEGG.AutoDock 1.5.6 was used to verify the molecular docking of the obtained target and curcumin.In an animal experiment,24 New Zealand white rabbits were selected and divided into blank groups,model groups,and high-dose and low-dose drug intervention groups.Eighteen rabbits were selected to establish a steroid-induced femoral head necrosis model induced by prednisolone acetate.Corresponding drug intervention was taken in each group,and HE staining,immunohistochemistry,and Western blot were used for detection,so as to compare the effect of TASDR on the expression of some key proteins in PI3K/AKT signal pathway during the treatment of steroid-induced femoral head necrosis.Results After screening,there were 38 potential core targets for TASDR to treat steroid-induced femoral head necrosis,including TNF,IL-6,VEGFA,etc.In addition,the target of TASDR included the AKT protein family AKT1.TASDR regulated osteoblastic differentiation through tumor signal pathway,VEGF signal pathway,and molecular functions such as enzyme binding and receptor binding and participated in biological processes such as programmed cell apoptosis and negative regulation of programmed cell apoptosis.Molecular docking showed that TASDR had a good binding activity with TNF,VEGF,AKT1,and other target genes.The results of animal experiments showed that in the model group,

关 键 词：网络药理学、分子对接、续断散加当归、骨碎补 激素性股骨头坏死 P13K/AKT信号通路 

分 类 号：R285.6[医药卫生—中药学]