目的 基于網(wǎng)絡(luò)藥理學(xué)及分子對接技術(shù)探究大黃治療膿毒癥的作用機制。方法 通過中藥系統(tǒng)藥理學(xué)數(shù)據(jù)庫與分析平臺(TCMSP)獲取大黃的有效活性成分及其作用靶點;應(yīng)用Cytoscape 3.9.1構(gòu)建大黃-活性成分-靶點的網(wǎng)絡(luò)圖;檢索GeneCards、OMIM、Drugbank、TTD數(shù)據(jù)庫,得到膿毒癥相關(guān)靶點;通過建立維恩圖獲得中藥與疾病交叉的關(guān)鍵靶點;應(yīng)用STRING平臺構(gòu)建關(guān)鍵靶點蛋白質(zhì)相互作用(PPI)網(wǎng)絡(luò)圖;使用DAVID數(shù)據(jù)庫對關(guān)鍵靶點進行基因本體(GO)生物學(xué)富集分析及京都基因與基因組百科全書(KEGG)通路富集分析。最后,應(yīng)用PyMOL軟件和AutoDock Vina軟件對大黃活性成分及其關(guān)鍵靶點進行分子對接驗證。結(jié)果 獲得大黃10種有效活性成分,其中重要的成分包括β-谷甾醇、蘆薈大黃素、澤蘭黃醇素等;藥物靶點與疾病靶點交集后獲得34個關(guān)鍵靶點,核心靶點有腫瘤壞死因子(TNF)、白細胞介素-1B(IL-1β)、腫瘤蛋白p53(TP53)、原癌基因(MYC)、半胱氨酸天冬氨酸蛋白酶-3(CASP3)、前列腺素內(nèi)過氧化物合酶2(PTGS2)、過氧化物酶體增殖物激活受體G(PPARG)、轉(zhuǎn)錄因子AP-1(JUN)、雌激素受體1(ESR1)、半胱氨酸蛋白酶8(CASP8)。GO富集分析表明大黃的關(guān)鍵靶點主要富集于306個生物過程、29個細胞組成和61個分子功能;KEGG通路富集分析確定了111條相關(guān)信號通路,包括磷脂酰肌醇-3-羥激酶(PI3K)-蛋白激酶B(Akt)信號通路、絲裂原活化蛋白激酶(MAPK)信號通路、糖尿病并發(fā)癥相關(guān)的晚期糖基化終末化產(chǎn)物(AGE)-晚期糖基化終末產(chǎn)物受體(RAGE)信號通路等;分子對接結(jié)果顯示,大黃的主要活性成分蘆薈大黃素與關(guān)鍵蛋白TNF、IL-1β、TP53、MYC、CASP3擁有良好的結(jié)合能力。結(jié)論 大黃通過多成分、多靶點和多通路治療膿毒癥。

Objective To explore the therapeutic effect and mechanism of Rhei Radix et Rhizoma on sepsis based on network pharmacology and molecular docking.Methods Active components and action targets of Rhei Radix et Rhizoma were obtained by using TCMSP database. The target network diagram “drug - component-target” of Rhei Radix et Rhizoma in treating sepsis was constructed by using Cytoscape 3.9.1. The sepsis disease targets were screened with GeneCards, OMIM, Drugbank, and TTD databases. The intersection gene targets were found by establishing Venn map of drug target genes and disease targets. The intersection genes were imported into STRING database to construct a protein-protein interaction(PPI) network. The genetic ontology(GO)biological process and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis were carried out through DAVID database. Finally, molecular docking technology was used to verify the result between ingredients and targets.Results A total of 10 active components of Rhei Radix et Rhizoma were obtained by TCMSP, including β-sitosterol, aloe-emodin, and eupatin.The number of targets that intersect with sepsis was 34. TNF, IL-1B, TP53, MYC, CASP3, PTGS2, PPARG, JUN, ESR1, CASP8 may be the key targets of rhubarb in treatment of sepsis. GO enrichment analysis showed that the key targets of Rhei Radix et Rhizoma were mainly enriched in 306 biological processes, 29 cell components and 61 molecular functions. KEGG pathway enrichment analysis identified 111 related signaling pathways, including PI3K-Akt signaling pathway, MAPK signaling pathway, and AGE-RAGE signaling pathway in diabetic complications. The results of molecular docking showed that aloe-emodin, the main active component of Rhei Radix et Rhizoma, had good binding ability with the key proteins TNF, IL-1β, TP53, MYC, and CASP3.Conclusions Rhei Radix et Rhizoma treats sepsis through multi-component, multi-target and multi-pathway treatment.

R914

上海市青年科技英才揚帆計劃(21YF1448300);上海市衛(wèi)生健康委中醫(yī)藥傳承和科技創(chuàng)新項目(ZYCC2019005)