基于网络药理学及实验验证研究治疗骨关节炎的潜在药物  

Exploration on potential drugs in treatment of osteoarthritis based on network pharmacology and experimental verification

作　　者：刘霞 周本宏[1] LIU Xia;ZHOU Benhong(Department of Pharmacy,Renmin Hospital of Wuhan University,Wuhan 430060,China)

机构地区：[1]武汉大学人民医院药学部,武汉430060

出　　处：《山西医科大学学报》2023年第8期1138-1146,共9页Journal of Shanxi Medical University

摘　　要：目的 运用网络药理学探索治疗骨关节炎(osteoarthritis,OA)的潜在药物并进行实验验证。方法 通过数据库及文献验证获取OA疾病靶点,根据候选靶点使用中药系统药理学数据库与分析平台(Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform,TCMSP)挖掘作用于以上靶点的化合物及包含化合物的中药,通过构建可视化网络,筛选核心靶点及化合物进行分子对接。根据分子对接结果选择化合物进行体外实验,选取大鼠关节软骨细胞,通过CellTiter-Glo(CTG)检测各化合物对软骨细胞活力影响,确定后续实验给药浓度。使用白细胞介素-1β(IL-1β)诱导软骨细胞建立OA细胞模型,将细胞分为对照组、OA组及给药组,使用流式细胞术检测化合物抗OA软骨细胞凋亡能力,并选择其中抗凋亡效果最显著的化合物,使用蛋白印迹法检测其对软骨细胞凋亡相关蛋白MDM2、P53、Bax表达水平的影响。结果 通过数据库检索、筛选及文献验证得到87个OA靶点,其中15个靶点匹配到符合药代动力学参数(ADME)和Lipinski筛选标准的化合物,并得到746个候选化合物,从数据库中筛选得到包含以上化合物的中药433种。通过靶点、化合物、中药可视化网络获得核心靶点前列腺素G/H合成酶2(PTGS2)、雌激素受体(ESR1)、肿瘤坏死因子(TNF)、白细胞介素-6(IL6)、IL-1β,核心化合物28个。以上核心靶点和核心化合物分子对接结果显示,C-3位为羟基的甾体化合物与核心靶点表现出良好的对接效果,根据分子对接结果从候选化合物中选择3种C-3位为羟基的甾体化合物菜油甾醇、豆甾醇、薯蓣皂苷元进行体外实验验证其治疗OA的效果。体外实验发现,与OA组比较,菜油甾醇组、豆甾醇组和薯蓣皂苷元组OA软骨细胞凋亡率显著下降(P<0.05),其中菜油甾醇抑制效果最佳。蛋白印迹法检测发现,与对照组比较,OA组MDM2蛋白表达降低,P53、Bax蛋白�Objective To explore the potential drugs for the treatment of osteoarthritis(OA)by network pharmacology,molecular docking and experimental verification.Methods OA disease targets were obtained through database and literature verification.Compounds acting on the above targets and Chinese medicines containing these compounds were screened using Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)according to the candidate targets.The core targets and the compounds were screened for molecular docking by constructing visual networks.According to the results of molecular docking,the compounds were selected for in vitro experiments.Rat articular chondrocytes were selected for the experiment,and the effect of compounds on chondrocyte viability was detected by CellTiter-Glo(CTG)to determine the dose concentration for the subsequent experiments.Chondrocytes were treated with interleukin-1 beta(IL-1β)to induce OA cell model,and the cells were divided into control group,OA group and drug groups.Flow cytometry was used to detect the anti-apoptotic ability of compounds in OA chondrocytes,and the compound with the most significant anti-apoptotic effect was selected.Then Western blotting was used to detect the effect of this compound on the expression levels of chondrocyte apoptosis-related proteins MDM2,P53 and Bax.Results A total of 87 OA targets were obtained through database mining,screening and literature verification,among which 15 targets were matched to compounds that met the screening criteria of ADME and Lipinski.A total of 746 candidate compounds were obtained,and 433 Chinese medicines containing these compounds were screened from the database.The core targets prostaglandin G/H synthase 2(PTGS2),estrogen receptor(ESR1),tumor necrosis factor(TNF),interleukin-6(IL6),IL-1βand 28 core compounds were obtained by the target-compound-Chinese medicine visual network.Molecular docking of the core targets and the core compounds showed that the C-3 hydroxyl steroid compounds had good docking

关 键 词：骨关节炎 网络药理学 分子对接 甾体化合物 菜油甾醇 软骨细胞 

分 类 号：R684.3[医药卫生—骨科学]