基于网络药理学技术探究雪莲注射液抗类风湿关节炎的作用及机制研究  

Mechanism of Xuelian injection for rheumatoid arthritis based on network pharmacology

作　　者：顾春宇 王亦巍 康建英 颜丽珊 邱新宇 罗赣[1] 张翼[1] GU Chun-yu;WANG Yi-wei;KANG Jian-ying;YAN Li-shan;QIU Xin-yu;LUO Gan;ZHANG Yi(School of Chinese Materia Medica,Beijing University of Chinese Medicine,Beijing 102488)

机构地区：[1]北京中医药大学中药学院,北京102488

出　　处：《中南药学》2025年第3期628-637,共10页Central South Pharmacy

基　　金：2017年度基本科研业务费项目(青年教师)(No.2017-JYB-JS-057)。

摘　　要：目的探究雪莲注射液(XLI)治疗类风湿关节炎(RA)的作用及其机制。方法采用网络药理学预测XLI治疗RA的活性成分、作用靶点和信号通路,选择活性成分与靶点基因通过分子对接技术进行验证。建立脂多糖(LPS)诱导的RAW264.7炎症细胞模型,给予不同浓度的XLI进行治疗。采用实时荧光定量PCR(qRT-PCR)检测炎症因子TNFA、IL^(-1)B和MCP-1基因的表达。采用蛋白免疫印迹法检测核心靶点COX2、eNOS、p-AKT和AKT的表达水平。建立弗氏完全佐剂(CFA)诱导的RA大鼠模型,给予不同浓度的XLI进行治疗并收集血清,采用非靶向代谢组学技术检测血清代谢物的水平。结果网络药理学筛选出XLI治疗RA的17个潜在活性成分和165个潜在作用靶点,蛋白互作网络核心靶点为炎性因子,GO和KEGG富集分析涉及MAPK、NF-κB、FoxO等多条信号通路。体外实验显示XLI能明显降低巨噬细胞TNFA、IL^(-1)B和MCP-1基因的表达,并降低COX2、eNOS和p-AKT蛋白表达水平。血清非靶向代谢组学显示XLI主要改变了花生四烯酸氧基蛋白代谢通路、鞘脂在内质网和高尔基体中的代谢、白三烯C4合成不足、罗非昔布和水杨酸钠药物的作用途径,并明显下调血清5,6-环氧二十碳三烯酸(5,6-EET)、12S-羟基十七碳三烯酸(12S-HHT)、血栓素B2(TXB2)、12-oxo-5Z,8Z,10E,14Z-二十碳四烯酸(12-KETE)、二氢鞘氨醇、3-脱氢鞘氨醇、8-异前列腺素A1(8-iso-PGA1)的水平。结论XLI治疗RA的作用机制可能与其抑制巨噬细胞TNFA、IL^(-1)B和MCP-1基因的表达,抑制COX2、eNOS和p-AKT蛋白的表达及花生四烯酸代谢通路有关。Objective To determine the mechanism of Xuelian injection(XLI)in the treatment of rheumatoid arthritis(RA).Methods The active ingredients,targets and signal pathways of XLI for RA were predicted with network pharmacology,and the active ingredients and target genes were verified by molecular docking.RAW264.7 cells were induced by lipopolysaccharide(LPS),and then treated with different concentrations of XLI.Real-time quantitative fluorescent PCR(qRT-PCR)was used to detect the gene expression of inflammatory cytokines,including TNFA,IL^(-1)B,and MCP-1.The expression levels of the core targets COX2,eNOS,p-AKT and AKT were detected by Western blot.The arthritis model of rat induced by Fraulein complete adjuvant(CFA)was established and then treated with different concentrations of XLI.The metabolites change in the serum were detected by non-targeted metabolomics.Results Totally 17 potential active ingredients and 165 potential targets of XLI for RA were screened by network pharmacology.The core targets of protein-protein interaction network were inflammation mediators.GO and KEGG enrichment analysis involved many signaling pathways,such as MAPK,NF-κB,and FoxO.The expression of TNFA,IL^(-1)B,and MCP-1,and the expression levels of COX2,eNOS and p-AKT proteins in LPS-induced RAW264.7 cells were decreased after the XLI treatment.Non-targeted metabolomics in the serum showed that XLI mainly changed the metabolic pathways of arachidonic acid oxy protein,sphingolipid metabolism in endoplasmic reticulum and Golgi apparatus,leukotriene C4 synthesis insufficiency,and metabolic pathways of rofecoxib,sodium salicylate action pathways in RA rats.XLI significantly reduced the serum levels of 5,6-EET,12S-HHT,thromboxane B2(TXB2),12-oxo-5Z,8Z,10E,14Z-eicosatetraenoic acid(12-KETE),dihydrosphingosine,3-dehydrosphingosine,and 8-isoprostaglandin A1(8-iso-PGA1).Conclusion The underlying mechanism of XLI for RA may be related to the suppression of XLI on TNFA,IL^(-1)B,and MCP-1 expression and expression of COX2,eNOS and p-AKT proteins,plu

关 键 词：雪莲注射液 类风湿关节炎 网络药理学 RAW264.7细胞 血清代谢组学 

分 类 号：R285[医药卫生—中药学]