基于网络药理学和实验验证白藜芦醇治疗类风湿性关节炎的作用机制

目的 通过网络药理学、分子动力学模拟探讨白藜芦醇对类风湿性关节炎的治疗作用及潜在机制，并进行实验验证。方法 使用TCMSP、Batman-TCM、SwissTarget Prediction数据库预测白藜芦醇的作用靶点；使用GeneCards和OMIM数据库预测类风湿性关节炎的潜在治疗靶点；使用STRING和Cytoscape3.8.0构建蛋白相互作用（PPI）网络，并进行MCC分析、MCODE分析、基因本体（GO）功能和京都基因与基因组百科全书（KEGG）富集分析，对KEGG通路和靶点之间绘制网络图；使用分子对接和分子动力学模拟方法对核心靶点与白藜芦醇进行模拟；构建类风湿性关节炎大鼠模型，Western blotting法检测p-磷脂酰肌醇3-激酶（p-PI3K）、p-蛋白激酶B1（p-Akt1）和p-细胞外信号调节激酶（p-ERK）1/2的表达水平。结果 白藜芦醇有310个靶点，与类风湿性关节炎有207个交集靶点，MCC分析发现Akt1、PIK3CA、PIK3CB、促分裂原活化蛋白激酶（MAPK）1、MAPK3、信号传导与转录激活因子3（STAT3）是其中核心基因。GO和KEGG富集分析发现，癌症通路、糖尿病并发症中的晚期糖基化终末产物–晚期糖基化终末产物受体（AGE-RAGE）信号通路、细胞凋亡和PI3K/Akt信号通路可能是白藜芦醇影响类风湿性关节炎的关键通路；分子对接和分子动力学模拟也发现Akt1、PIK3CA和MAPK1与白藜芦醇之间有很强的结合能力。动物实验发现，与对照组相比，模型组大鼠关节炎指数明显增加，而给予白藜芦醇后大鼠的关节炎指数明显下降；p-PI3K、p-AKT表达明显上升，p-ERK1/2的表达明显下降（P＜0.05、0.01）。结论 白藜芦醇能改善类风湿性关节炎大鼠的关节炎指数，并可能通过Akt1、PIK3CA和MAPK1等靶点和PI3K/Akt信号通路来发挥关键治疗作用。

Mechanism of action of resveratrol in treatment of rheumatoid arthritis through network pharmacology and experimental validation

Objective To explore the therapeutic effects and potential mechanisms of resveratrol in rheumatoid arthritis through network pharmacology, molecular dynamics simulations and animal experiments, and experimental verification was carried out. Methods The TCMSP, Batman-TCM, and SwissTargetPrediction databases were used to predict the targets of action of resveratrol. GeneCards and OMIM databases were used to predict the potential therapeutic targets of rheumatoid arthritis. PPI networks were constructed using STRING and Cytoscape 3.8.0, and MCC analysis, MCODE analysis, GO and KEGG enrichment analysis, and target network mapping between KEGG pathways and targets were performed. Using molecular docking and molecular dynamics simulation methods to simulate the core targets with resveratrol. Constructing a rheumatoid arthritis rat model, and detecting the expression levels of p-PI3K, p-Akt, and p-ERK1/2 by Western blotting. Results A total of 310 targets of resveratrol and 207 intersecting targets with rheumatoid arthritis. MCC analysis found that Akt1, PIK3CA, PIK3CB, MAPK1,MAPK3, and STAT3 are the core genes among them. GO and KEGG enrichment analysis found that pathways in cancer, AGE-RAGE signaling pathways in cancer, apoptosis, PI3K-Akt signaling pathway may be the key pathways of resveratrol affecting rheumatoid arthritis. Molecular docking and molecular dynamics simulation also found that Akt1, PIK3CA, and MAPK1 have strong binding ability with resveratrol. Animal experiments revealed that compared with the control group, the arthritis index of the model group was significantly increased, but the arthritis index in the resveratrol group was significantly decreased. The expression of p-PI3K and p-Akt was significantly increased, while the expression of p-ERK1/2 was significantly decreased (P < 0.05, 0.01). Conclusion Resveratrol can improve arthritis index of rheumatoid arthritis rats, and may play a key therapeutic role through the targets of Akt1, PIK3CA, and MAPK1 and the PI3K/Akt signaling pathway.