目的 基于網(wǎng)絡(luò)藥理學(xué)和分子對(duì)接技術(shù)探討補(bǔ)腎復(fù)方五子衍宗丸治療肝纖維化的作用機(jī)制。方法 使用中藥系統(tǒng)藥理學(xué)數(shù)據(jù)庫(kù)與分析平臺(tái)(TCMSP,http://lsp.nwu.edu.cn/tcmspsearch.php)數(shù)據(jù)庫(kù)預(yù)測(cè)五子衍宗丸有效成分的靶點(diǎn)并通過(guò)Uniprot數(shù)據(jù)庫(kù)進(jìn)行靶點(diǎn)名稱(chēng)標(biāo)準(zhǔn)化;通過(guò)OMIM、Gene Cards、DisGeNET數(shù)據(jù)庫(kù)收集肝纖維化靶點(diǎn),通過(guò)韋恩圖篩選活性成分靶點(diǎn)和疾病靶點(diǎn)基因的交集,獲取五子衍宗丸治療肝纖維化的潛在靶點(diǎn);運(yùn)用Cytoscape 3.9.0構(gòu)建有效成分-靶點(diǎn)-肝纖維化網(wǎng)絡(luò),預(yù)測(cè)核心靶點(diǎn),構(gòu)建基于STRING平臺(tái)的蛋白質(zhì)-蛋白質(zhì)相互作用(PPI)網(wǎng)絡(luò);利用微生信平臺(tái)進(jìn)行基因本體(GO)功能和京都基因與基因組百科全書(shū)(KEGG)通路富集分析;使用AutoDockTools軟件,結(jié)合PPI網(wǎng)絡(luò)及GO功能、KEGG通路分析,將核心靶點(diǎn)與有效成分進(jìn)行對(duì)接驗(yàn)證。結(jié)果 獲得五子衍宗丸活性成分84個(gè),篩選出有效成分作用靶點(diǎn)179個(gè),肝纖維化靶點(diǎn)798個(gè),五子衍宗丸與肝纖維化交集靶點(diǎn)81個(gè),經(jīng)拓?fù)鋵傩苑治龊Y選得到五子衍宗丸26個(gè)活性成分、37個(gè)治療肝纖維化關(guān)鍵靶點(diǎn);根據(jù)“活性成分-靶點(diǎn)-通路”網(wǎng)絡(luò)圖,預(yù)測(cè)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(xì)胞介素-6(IL6)、細(xì)胞腫瘤抗原p53(TP53)、血管內(nèi)皮生長(zhǎng)因子A(VEGFA)等為核心靶點(diǎn)。GO功能富集分析的生物學(xué)過(guò)程(BP)主要與RNA聚合酶II啟動(dòng)子轉(zhuǎn)錄的正調(diào)控、基因表達(dá)的正調(diào)控等關(guān)聯(lián);細(xì)胞組分(CC)主要與細(xì)胞外空間、細(xì)胞核等關(guān)系密切;分子功能(MF)主要與大分子復(fù)合物結(jié)合、蛋白激酶活性、RNA聚合酶II核心啟動(dòng)子近端序列特異性DNA結(jié)合等相關(guān)。五子衍宗丸治療肝纖維化途徑主要包括化學(xué)致癌-活性氧信號(hào)通路、JAK-STAT信號(hào)通路、PI3K-Akt信號(hào)通路、MAPK信號(hào)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(diǎn)AKT1、TNF、IL6、VEGFA、CASP3等對(duì)接結(jié)合穩(wěn)定,分子對(duì)接初步證明關(guān)鍵成分自發(fā)地與多個(gè)核心蛋白結(jié)合,可對(duì)多個(gè)關(guān)鍵靶點(diǎn)進(jìn)行調(diào)控。結(jié)論 通過(guò)網(wǎng)絡(luò)藥理學(xué)及分子對(duì)接初步揭示補(bǔ)腎復(fù)方五子衍宗丸影響凋亡過(guò)程的正負(fù)調(diào)控、氧化應(yīng)激、炎癥因子、血管生成等反應(yīng)過(guò)程,具有多成分、多靶點(diǎn)、多通路等作用特點(diǎn),可為臨床肝纖維化治療中重視補(bǔ)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(chǔ)與應(yīng)用基礎(chǔ)項(xiàng)目(202201010183); 廣東省中醫(yī)藥局科研項(xiàng)目(20222112)