藏药如意珍宝丸治疗退行性骨关节炎的作用机理研究  

Study on the mechanism of Zang Medicine Ruyi Zhenbao Pill in treating degenerative osteoarthritis

作　　者：周辛萍 申李丹 袁发荣 黄先菊[5] 郭肖 李啟恩 ZHOU Xinping;SHEN Lidan;YUAN Farong;HUANG Xianju;GUO Xiao;LI Qien(Qinghai University,State Key Laboratory of Zang Medicine Research and Development,Xining 810016,China;Qinghai University,Zang Medicine Research Center,Xining 810016,China;Qinghai University,Zang Medical College,Xining 810016,China;Medical Education Department of Maternal and Child Health Hospital of Qinghai Province,Xining 810016,China;School of Pharmaceutical Sciences,South-Central Minzu University,Wuhan 430074,China;Arura Zang Medicine Co.,Ltd.,Xining 810016,China)

机构地区：[1]青海大学藏医学院,西宁810016 [2]青海大学藏医药研究中心,西宁810016 [3]青海大学藏药新药开发国家重点实验室,西宁810016 [4]青海省妇幼保健医院医务科,西宁810016 [5]中南民族大学药学院,武汉430074 [6]金诃藏药股份有限公司,西宁810016

出　　处：《中南民族大学学报（自然科学版）》2024年第6期742-752,共11页Journal of South-Central Minzu University(Natural Science Edition)

基　　金：青海省重大科技专项资助项目(2020-SF-A3-2);国家中医药领军人才支持计划“青年岐黄学者”项目。

摘　　要：基于网络药理学、分子对接及体外实验来探讨如意珍宝丸(Ruyi Zhenbao Pill,RZP)治疗退行性骨关节炎(degenerative osteoarthrosis,DOA)的作用机制.通过数据库检索并筛选得到RZP活性成分及DOA相关靶点,构建蛋白质互作网络图,绘制药物-成分-核心靶点-疾病网络图及核心成分-靶点网络图;对共同靶点进行KEGG富集分析;将获得的RZP主要活性成分与核心靶点进行分子对接验证.以脂多糖(lipopolysaccharide,LPS)诱导构建ATDC5细胞炎症模型,采用CCK-8法和qRT-PCR方法分别验证细胞活力及相关mRNA的表达量.最后得到RZP成分共1602个,交集靶点共31个;KEGG富集分析得到类风湿性关节炎通路、破骨细胞分化通路等信号通路共33条;分子对接结果显示β-谷甾醇与对接的核心靶点具有较高的亲和力;体外细胞实验表明,给予RZP后,ATDC5细胞活力较模型组明显上升,RZP可调节TNF-α、iNOS、COX-2、IL-6靶点mRNA的表达.如意珍宝丸可能通过多成分、多靶点和多途径协同作用治疗DOA的药理特点,为其藏医临床用药及大品种培育提供了科学基础.Aim to study the mechanism of action of Ruyi Zhenbao Pill in the treatment of degenerative osteoarthritis(DOA)based on network pharmacology,molecular docking and experimental validation.The active ingredients targets of RZP and disease targets of DOA were collected through databases.The common targets were utilized by mixture of them and the core targets were obtained by constructing the protein-protein interaction(PPI)network.Then the drug-componentcore targets-disease network diagram and core component-targets diagram were constructed.The gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)pathway enrichment analysis were carried.AutoDock Vina software was utilized for combining the core targets and the key ingredients of RZP.Finally,ATDC5 cells were used to construct a cellular inflammation model with lipopolysaccharide,cell viability was assessed by CCK-8 assay,and the mRNA expression of relevant core targets was assessed by qRT-PCR.A total 1602 ingredients were searched from databases,and 31 common targets were obtained.Altogether 31 pathways were screened by KEGG enrichment,mainly including Rheumatoid Arthritis Pathway and Osteoclast differentiation pathway.Molecular docking results indicated that the beta-sitosterol of RAP had a high affinity with the core target to be docked.In vitro experiment suggested that the activity of ATDC5 cells increased significantly compared with the model group.RZP could regulate the mRNA expression of TNF-α,iNOS,COX-2 and IL-6.The synergistic effects of RZP in treating DOA with multi-component,multi-target and multi-pathway,and also provides a pre-experimental basis for clinical application and new drug development.

关 键 词：如意珍宝丸 退行性骨关节炎 网络药理学 分子对接 体外验证 

分 类 号：R285.5[医药卫生—中药学]