目的 通過網(wǎng)絡(luò)藥理學、分子動力學模擬探討白藜蘆醇對類風濕性關(guān)節(jié)炎的治療作用及潛在機制，并進行實驗驗證。方法 使用TCMSP、Batman-TCM、SwissTarget Prediction數(shù)據(jù)庫預測白藜蘆醇的作用靶點；使用GeneCards和OMIM數(shù)據(jù)庫預測類風濕性關(guān)節(jié)炎的潛在治療靶點；使用STRING和Cytoscape3.8.0構(gòu)建蛋白相互作用（PPI）網(wǎng)絡(luò)，并進行MCC分析、MCODE分析、基因本體（GO）功能和京都基因與基因組百科全書（KEGG）富集分析，對KEGG通路和靶點之間繪制網(wǎng)絡(luò)圖；使用分子對接和分子動力學模擬方法對核心靶點與白藜蘆醇進行模擬；構(gòu)建類風濕性關(guān)節(jié)炎大鼠模型，Western blotting法檢測p-磷脂酰肌醇3-激酶（p-PI3K）、p-蛋白激酶B1（p-Akt1）和p-細胞外信號調(diào)節(jié)激酶（p-ERK）1/2的表達水平。結(jié)果 白藜蘆醇有310個靶點，與類風濕性關(guān)節(jié)炎有207個交集靶點，MCC分析發(fā)現(xiàn)Akt1、PIK3CA、PIK3CB、促分裂原活化蛋白激酶（MAPK）1、MAPK3、信號傳導與轉(zhuǎn)錄激活因子3（STAT3）是其中核心基因。GO和KEGG富集分析發(fā)現(xiàn)，癌癥通路、糖尿病并發(fā)癥中的晚期糖基化終末產(chǎn)物–晚期糖基化終末產(chǎn)物受體（AGE-RAGE）信號通路、細胞凋亡和PI3K/Akt信號通路可能是白藜蘆醇影響類風濕性關(guān)節(jié)炎的關(guān)鍵通路；分子對接和分子動力學模擬也發(fā)現(xiàn)Akt1、PIK3CA和MAPK1與白藜蘆醇之間有很強的結(jié)合能力。動物實驗發(fā)現(xiàn)，與對照組相比，模型組大鼠關(guān)節(jié)炎指數(shù)明顯增加，而給予白藜蘆醇后大鼠的關(guān)節(jié)炎指數(shù)明顯下降；p-PI3K、p-AKT表達明顯上升，p-ERK1/2的表達明顯下降（P＜0.05、0.01）。結(jié)論 白藜蘆醇能改善類風濕性關(guān)節(jié)炎大鼠的關(guān)節(jié)炎指數(shù)，并可能通過Akt1、PIK3CA和MAPK1等靶點和PI3K/Akt信號通路來發(fā)揮關(guān)鍵治療作用。

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.

R681

江蘇省衛(wèi)生健康委員會重點項目（ZD2022064）；江蘇省中醫(yī)藥科技發(fā)展項目（MS2021102）