目的 基于網絡藥理學和分子對接技術探討補腎復方五子衍宗丸治療肝纖維化的作用機制。方法 使用中藥系統(tǒng)藥理學數(shù)據(jù)庫與分析平臺(TCMSP,http://lsp.nwu.edu.cn/tcmspsearch.php)數(shù)據(jù)庫預測五子衍宗丸有效成分的靶點并通過Uniprot數(shù)據(jù)庫進行靶點名稱標準化;通過OMIM、Gene Cards、DisGeNET數(shù)據(jù)庫收集肝纖維化靶點,通過韋恩圖篩選活性成分靶點和疾病靶點基因的交集,獲取五子衍宗丸治療肝纖維化的潛在靶點;運用Cytoscape 3.9.0構建有效成分-靶點-肝纖維化網絡,預測核心靶點,構建基于STRING平臺的蛋白質-蛋白質相互作用(PPI)網絡;利用微生信平臺進行基因本體(GO)功能和京都基因與基因組百科全書(KEGG)通路富集分析;使用AutoDockTools軟件,結合PPI網絡及GO功能、KEGG通路分析,將核心靶點與有效成分進行對接驗證。結果 獲得五子衍宗丸活性成分84個,篩選出有效成分作用靶點179個,肝纖維化靶點798個,五子衍宗丸與肝纖維化交集靶點81個,經拓撲屬性分析篩選得到五子衍宗丸26個活性成分、37個治療肝纖維化關鍵靶點;根據(jù)“活性成分-靶點-通路”網絡圖,預測槲皮素、山柰酚、苦參堿、黃豆黃素為五子衍宗丸作用于肝纖維化的重要活性成分;RAC-α絲氨酸/蘇氨酸蛋白激酶(AKT1)、腫瘤壞死因子(TNF)、白細胞介素-6(IL6)、細胞腫瘤抗原p53(TP53)、血管內皮生長因子A(VEGFA)等為核心靶點。GO功能富集分析的生物學過程(BP)主要與RNA聚合酶II啟動子轉錄的正調控、基因表達的正調控等關聯(lián);細胞組分(CC)主要與細胞外空間、細胞核等關系密切;分子功能(MF)主要與大分子復合物結合、蛋白激酶活性、RNA聚合酶II核心啟動子近端序列特異性DNA結合等相關。五子衍宗丸治療肝纖維化途徑主要包括化學致癌-活性氧信號通路、JAK-STAT信號通路、PI3K-Akt信號通路、MAPK信號通路等。核心成分槲皮素、山柰酚、苦參堿、黃豆黃素與核心作用靶點AKT1、TNF、IL6、VEGFA、CASP3等對接結合穩(wěn)定,分子對接初步證明關鍵成分自發(fā)地與多個核心蛋白結合,可對多個關鍵靶點進行調控。結論 通過網絡藥理學及分子對接初步揭示補腎復方五子衍宗丸影響凋亡過程的正負調控、氧化應激、炎癥因子、血管生成等反應過程,具有多成分、多靶點、多通路等作用特點,可為臨床肝纖維化治療中重視補腎法提供參考依據(jù)。

Objective The mechanism of the kidney-tonifying formula Wuzi Yanzong Pills was explored in treatment of hepatic fibrosis by means of network pharmacology and molecular docking technology. Method The active components of Wuzi Yanzong Pills and it's corresponding targets were predicted by Using TCMSP database and the target names were standardized through the Uniprot database. The hepatic fibrosis targets were collected through OMIM, Gene Cards, and DisGeNET databases, the potential targets of Wuzi Yanzong Pills in treatment of liver fibrosis were obtained by the intersection of active component targets and disease targets selected by Venn diagram. Cytoscape 3.9.0 software was used to map the active components-intersection targets-disease network and to predict core targets. STRING platform was used to construct a PPI network. Pathway enrichment was analyzed by gene ontology(GO) function and Kyoto encyclopedia of geneses and genomes(KEGG); AutoDockTools software was used to dock the core target with active components. Result 84 active components of Wuzi Yanzong Pills and 179 potential targets of the active components were obtained, 798 hepatic fibrosis targets and 81 intersection targets between the disease and Wuzi Yanzong Pills were identified. 26 active components and 37 key targets for the treatment of hepatic fibrosis by Wuzi Yanzong Pills were screened by meridian topology analysis. Quercetin, kaempferol, matrine, and glycitein were important active components of Wuzi Yanzong Pills in treating hepatic fibrosis which were predicted by the "active ingredient target pathway" network. The core targets were RAC-αSerine/threonine protein kinase(AKT1), tumor necrosis factor(TNF), interleukin-6(IL6), cell tumor antigen p53(TP53), vascular endothelial growth factor A(VEGFA), etc. The biological process of GO functional enrichment analysis were mainly related to the positive regulation of RNA polymerase II promoter transcription and gene expression, the cellular components were closely related to the extracellular space, nucleus, etc., the molecular function was mainly related to the binding of macromolecular complexes,protein kinase activity, and specific DNA binding to the proximal sequence of RNA polymerase II core promoter. The main pathways through which Wuzi Yanzong Pills treated hepatic fibrosis include the chemical carcinogenic reactive oxygen species signaling pathway, JAK-STAT signaling pathway, PI3K-Akt signaling pathway, MAPK signaling pathway, etc. The core components of quercetin, kaempferol, matrine, and glycitein had stable docking and binding with core targets such as AKT1, TNF, IL6, VEGFA,CASP3, etc. Molecular docking had preliminarily demonstrated that key components spontaneously bind to multiple core proteins and can regulate multiple key targets.Conclusion The mechanism of the kidney-tonifying formula Wuzi Yanzong Pills in treatment of hepatic fibrosis was preliminarily revealed that it could interfere the the process of apoptosis through positive and negative regulation, oxidative stress, inflammatory factors, angiogenesis, and other response; The kidney tonifying formula Wuzi Yanzong Pills had the characteristics of multiple components, multiple targets, and multiple pathways, which can provide a reference basis for emphasizing the kidney tonifying method in the clinical treatment of hepatic fibrosis.

R285.5

廣州市科技局基礎與應用基礎項目(202201010183); 廣東省中醫(yī)藥局科研項目(20222112)