摘要
通过网络药理学分析及细胞实验,深入探究益肾壮筋合剂治疗骨质疏松症的作用机制,丰富益肾壮筋合剂治疗骨质疏松症的依据。通过TCMSP、HERB数据库检索益肾壮筋合剂所含中药的有效化学成分和作用靶点,以“osteoporosis”为检索词通过GeneCards、OMIM、DisGeNET数据库检索疾病靶点,使用Venny平台筛选出交集靶点。使用Cytoscape软件构建益肾壮筋合剂治疗骨质疏松症的药物-有效成分-靶点网络图,并在此基础上深入筛选,确定六大核心靶点。利用DAVID数据库进行GO功能富集分析以及KEGG信号通路分析,之后使用中药干预体外细胞成骨及破骨分化过程,通过基因测序与网络药理学结果进行对比分析。通过网络药理学方法共找到益肾壮筋合剂活性成分共937种,骨质疏松症对应靶点共计2 034个。通过Venny平台筛选到267个交集靶点,进一步通过软件分析得到排名前6的靶点为:AKT1、JUN、IL-6、STAT3、MAPK3、TNF。GO通路富集分析,得到575个相关通路,主要涉及酶结合、转录翻译过程、细胞器、细胞膜功能等方面。KEGG通路富集分析,结果表明核心靶点涉及激素反应、蛋白质反应、脂质代谢、氧代谢等信号通路,并得到主要4个功能聚类基团,并推测聚类基团1起主要作用。在体外实验部分,通过含药血清干预MC3T3-E1细胞成骨分化、Raw264.7细胞系破骨分化,以细胞测序结果进行对比,得到19个破骨相关基因和36个成骨相关基因,再与以网络药理学推测的聚类团1中包含的关键基因进行合并,进一步验证了IL-6、TNF是益肾壮筋合剂调控骨代谢的关键基因。研究结果初步预测了益肾壮筋合剂主要通过调控IL-6抑制破骨代谢,并通过TNF调控成骨代谢,起到治疗骨质疏松的作用。
Through network pharmacological analysis and cell experiment,the mechanism of Yishenzhuangjin mixture in treating osteoporosis was deeply explored,and the basis of Yishenzhuangjin mixture in treating osteoporosis was enriched.The effective chemical components and the target of TCM in Yishenzhuangjin mixture were searched through TCMSP and HERB databases,and the disease targets of“osteoporosis”were searched through GeneCards,OMIM and DisGeNET databases,and the intersections were selected by Venny platform.Cytoscape software was used to construct the drug-active ingredient-target network diagram of Yishenzhuangjin mixture for the treatment of osteoporosis,and on this basis,six core targets were determined.GO functional enrichment analysis and KEGG signal pathway analysis were performed using DAVID database.After that,Chinese medicine was used to intervene in the process of osteogenesis and osteoclast differentiation in vitro,and the results of gene sequencing and network pharmacology were compared and analyzed.A total of 937 active ingredients of Yishenzhuangjin mixture were identified through network pharmacology methods,and 2034 corresponding targets were identified for osteoporosis.267 intersecting targets were screened through the Venny platform,and further software analysis revealed the top 6 targets as AKT1,JUN,IL-6,STAT3,MAPK3,and TNF.GO pathway enrichment analysis identified 575 related pathways,mainly involving enzyme binding,transcription and translation processes,organelles,and membrane functions.KEGG pathway enrichment analysis showed that the core target involves signaling pathways such as hormone response,protein response,lipid metabolism,and oxygen metabolism,and identified four main functional clustering groups.Cluster group 1 was speculated to play a major role.In the in vitro experiment,the osteogenic differentiation of MC3T3-E1 cells and Raw264.7 cell line was intervened by drug containing serum.The results of cell sequencing were compared to obtain 19 osteoclast related genes and 36 osteogenic related genes.These genes were then combined with the key genes included in cluster 1 speculated by network pharmacology to further verify that IL-6 and TNF are key genes regulating bone metabolism in Yishenzhuangjin mixture.The research results preliminarily predict that Yishenzhuangjin mixture mainly inhibits osteoclast metabolism by regulating IL-6 and regulates osteogenic metabolism by TNF,playing a role in treating osteoporosis.
作者
陆苇
缪逸鸣
陈俊
江文涛
王强
蒋晓伟
LU Wei;MIAO Yiming;CHEN Jun;JIANG Wentao;WANG Qiang;JIANG Xiaowei(Department of Orthopedics,Nanjing University of Traditional Chinese Medicine Changshu Affiliated Hospital,Suzhou 215500,China)
出处
《药物生物技术》
2025年第5期591-599,共9页
Pharmaceutical Biotechnology
基金
苏州市科技计划项目(No.SYW2024151)
2019年度苏州市临床重点病种诊疗技术专项项目(No.LCZX201923)。
关键词
益肾壮筋合剂
骨质疏松症
网络药理学
体外实验
骨稳态
破骨代谢
成骨代谢
Yishenzhuangjin mixture
Osteoporosis
Network pharmacology
In vitro experiments
Bone homeostasis
Osteoclastic metabolism
Osteogenic metabolism
