基于系统药理学结合LC-MS/MS技术探讨温性经筋通贴膏治疗类风湿性关节炎的药效物质基础与作用特点

Exploration of the Pharmacological Substance Basis and Action Characteristics of Wenxing Jingjintong Gel Patch in the Treatment of Rheumatoid Arthritis Based on System Pharmacology and LC-MS/MS Technology

目的通过液相色谱-质谱-质谱联用(LC-MS/MS)技术结合系统药理学研究温性经筋通贴膏治疗类风湿性关节炎的药效物质基础与作用特点。方法通过LC-MS/MS技术结合文献检索获取温性经筋通贴膏活性成分信息,使用Swiss Target Prediction平台进行作用靶点预测;利用GeneCards、DrugBank和OMIM数据库检索与类风湿性关节炎相关的疾病靶点并与活性成分作用靶点进行映射取交集,通过Cytoscape软件构建“活性成分-有效靶点”网络;将共同靶点导入STRING数据库构建蛋白互作网络;应用Metascape数据库进行GO功能及KEGG通路富集分析;采用AutoDock软件进行关键成分与有效靶点的分子对接研究。结果通过应用LC-MS/MS技术从温性经筋通贴膏中共鉴别出142个活性成分,经检索相关文献补充活性成分至174个,与类风湿性关节炎交集靶点175个;预测温性经筋通贴膏可能通过小檗碱、新补骨脂异黄酮、盐酸巴马汀等主要成分,作用于TNF、IL6、AKT1等关键靶点,调控PI3K/Akt1、JAK/STAT、MAPK等信号通路发挥免疫调节与抗炎镇痛作用;在1152组分子对接验证中,结合能小于-5.0 kcal·mol^(-1)的占94%、小于-7.0 kcal·mol^(-1)的占51%,表明所筛选的潜在活性成分与核心靶点之间具有较好的结合亲和力。结论该研究预测了温性经筋通贴膏治疗类风湿性关节炎的药效物质基础与作用特点,为进一步启发其临床应用及质量控制提供了理论基础。

Objective To explore the key active ingredients and action characteristics of Wenxing Jingjintong Gel Patch in the treatment of rheumatoid arthritis(RA)based on the systematic pharmacology and LC-MS/MS technology.Methods The information of active ingredient from Wenxing Jingjintong Gel Patch was established through LC-MS/MS analysis and literature retrieval.The targets of the active ingredients were predicted using Swiss Target Prediction platform and then mapped with the RA-related targets obtained from GeneCards,DrugBank,and OMIM databases to identify the intersecting targets.The“active ingredients-effective targets”network was constructed through the Cytoscape software.The shared targets were imported into STRING database to construct a protein-protein interaction network.GO function and KEGG pathway enrichment analysis were performed using the Metascape database.Molecular docking studies were conducted using AutoDock software to investigate the interactions between key ingredients and target proteins.Results A total of 142 active ingredients were identified in Wenxing Jingjintong Gel Patch by wsing LC-MS/MS,which were further supplemented to 174 through literature retrieval.There were 175 shared targets between the active ingredients and RA.It was anticipated that Wenxing Jingjintong Gel Patch exerted immune regulation and anti-inflammatory and analgesic effects through the interaction between key active ingredients such as berberine,neobavaisoflavone,and palmatine chloride with key targets,including TNF,IL6,and AKT1 to regulate PI3K/Akt1,JAK/STAT,and MAPK signaling pathways.In 1152 molecular docking validation,94%of them had binding energies less than-5.0 kcal·mol^(-1),while 51%of them had binding energies less than-7.0 kcal·mol^(-1).It was indicated that there was a good binding affinity between the potential active ingredients and core targets.Conclusion This study predicted the active ingredients and action characteristics of Wenxing Jingjintong Gel Patch in the treatment of RA,which provided a theoretical basis for further clinical application and quality control.