当归补血汤对类风湿关节炎的干预作用及相关网络药理学研究

Interventional Effect and Network Pharmacological Study of Danggui Buxue Decoction on RheumatoidArthritis

目的利用网络药理学、分子对接技术结合实验验证探究当归补血汤治疗类风湿关节炎(Rheumatoid Arthritis,RA)的潜在作用机制。方法通过网络药理学相关数据库与分析平台获取当归补血汤的活性成分和作用靶点以及RA疾病靶点,通过STRING数据库绘制PPI蛋白互作图,分析当归补血汤治疗RA的核心靶点。通过DAVID数据库对交集靶点进行GO和KEGG富集分析,将药物主要活性成分与核心靶点进行分子对接初步验证。利用牛Ⅱ型胶原乳剂诱导建立类风湿关节炎大鼠模型,进行关节炎评分,Western Blot法检测关键蛋白进行实验验证。结果共筛选出当归补血汤主要活性成分17个,对应靶点193个,疾病靶点2883个,两者取交集后获得129个当归补血汤治疗RA的潜在靶点。分析得到AKT1、VEGFA、IL-6、HIF-1α、MMP9等27个核心靶点,918条GO相关条目,171条KEGG通路,包括PI3K-Akt信号通路、AGE-RAGE信号通路、TNF信号通路、MAPK等信号通路。分子对接显示HIF-1α、VEGFA、MMP9蛋白受体与药物主要活性成分槲皮素配体亲和力良好。动物实验验证结果表明,与空白组比较,模型组大鼠关节炎评分、关节滑膜中VEGFA、HIF-1α和MMP9关键蛋白表达均明显升高(P<0.01);与模型组比较,治疗组各组大鼠关节炎症状均明显改善,VEGFA、HIF-1α和MMP9蛋白表达均明显降低(P<0.05,P<0.01)。结论利用网络药理学-分子对接结合药效学实验,初步揭示了当归补血汤可能作用于VEGFA、HIF-1α和MMP9等多个靶点来干预RA的疾病进程,对RA起到治疗作用。

Objective To explore the potential mechanism of Danggui Buxue Decoction(DGBXD)in the treatment of rheumatoid arthritis(RA)by a combined approach of network pharmacology,molecular docking technology and experimental verification.Methods The active components and targets of DGBXD and RA disease targets were obtained through network pharmacology-related database and analysis platform.The protein-protein(PPI)interaction map was drawn by STRING database and core targets of DGBXD in the treatment of RA were analyzed.DAVID database was used to perform GO and KEGG enrichment analysis on the intersection targets.After that,molecular docking between the active components and the core target was conducted to make preliminary verification.A rat model of RA induced by bovine typeⅡcollagen emulsion was established.The degree of arthritis was assessed by visual scoring.Western Blot was used to detect key proteins to make experimental verfication.Results In this study,17 main active components of DGBXD,which corresponded to 193 targets and 2883 disease targets,were screened.After the intersection of two targets,129 potential targets of DGBXD in the treatment of RA were obtained.Twenty-seven core targets such as AKT1,VEGFA,IL-6,HIF-1α,MMP9,were obtained by topological analysis.A total of 918 GO-related items and 171 KEGG pathways,mainly involving PI3K-Akt signaling pathway,AGE-RAGE signaling pathway,TNF signaling pathway,MAPK and other signaling pathways were gained by enrichment analysis.Molecular docking results showed that the active component(quercetin)had good affinity with HIF-1α,VEGFA and MMP9.The experimental results showed that compared with the blank group,the arthritis score,and the key protein expressions of MMP9,VEGFA and HIF-1αin the synovium of the model group rats were significantly increased(P<0.01).Compared with the model group,the arthritis symptoms of rats in the treatment group were significantly improved,the protein expressions of MMP9,VEGFA and HIF-1αwere significantly decreased(P<0.05,P<0.01).Conclusion The combination of network pharmacology,molecular docking and pharmacodynamics experiment preliminarily revealed that DGBXD may act on VEGFA,HIF-1αand MMP9 to interfere with the disease process of RA and play a therapeutic role in RA.