目的 基于網(wǎng)絡(luò)藥理學(xué)和動物實(shí)驗(yàn)探究王不留行黃酮苷（VAC）對膿毒癥小鼠急性肺損傷的保護(hù)作用機(jī)制。方法 通過 Swiss Target Prediction數(shù)據(jù)庫收集 VAC的潛在作用靶點(diǎn)；利用 GeneCards數(shù)據(jù)庫檢索肺膿毒癥相關(guān)的疾病靶點(diǎn)；運(yùn)用Draw Venn Diagram 軟件構(gòu)建 VAC 和疾病的共同靶點(diǎn)；利用 STRING 11.5數(shù)據(jù)庫和 Cytoscape 3.10.0軟件構(gòu)建共同靶點(diǎn)蛋白質(zhì)-蛋白質(zhì)相互作用（PPI）網(wǎng)絡(luò)；利用 Metascape 數(shù)據(jù)庫進(jìn)行基因本體（GO）富集分析和京都基因與基因組百科全書（KEGG）通路分析。采用 ip 給予脂多糖（LPS）構(gòu)建膿毒癥模型，造模同時給予 VAC（1、5 mg·kg^-1）或地塞米松干預(yù)，取各組小鼠肺組織及血清，蘇木精-伊紅（HE）、Masson 及 TUNEL 染色觀察肺組織形態(tài)變化、纖維化及細(xì)胞凋亡情況，ELISA 法和實(shí)時熒光定量 PCR（qRT-PCR）法分別檢測血清和肺組織中炎癥因子水平，免疫組織化學(xué)法和 Western blotting法檢測肺組織炎癥通路相關(guān)蛋白表達(dá)。結(jié)果 VAC和肺膿毒癥共同靶點(diǎn)有44個；GO富集分析涉及生物過程（BP）823個條目、細(xì)胞組分（CC）52個條目和分子功能（MF）49個條目；KEGG分析篩選出癌癥通路、PI3K-Akt、JAK-STAT信號通路等 20 條信號通路。驗(yàn)證實(shí)驗(yàn)結(jié)果顯示，與對照組相比，模型組小鼠肺組織損傷且纖維化嚴(yán)重，肺臟指數(shù)顯著增加（P＜0.05），血清及肺組織中相關(guān)炎癥因子表達(dá)升高（P＜0.01、0.001）。與模型組相比，VAC及地塞米松組肺組織病理形態(tài)得到改善，纖維化程度減輕，肺臟指數(shù)顯著降低（P＜0.05），血清中腫瘤壞死因子（TNF-α）、白細(xì)胞介素-1β（IL-1β）、白細(xì)胞介素6（IL-6）水平、肺組織炎癥蛋白表達(dá)量及細(xì)胞凋亡數(shù)量降低，PI3K、Akt蛋白表達(dá)升高（P＜0.05、0.01、0.001）。動物實(shí)驗(yàn)結(jié)果與網(wǎng)絡(luò)藥理學(xué)結(jié)果一致。結(jié)論 VAC 對膿毒癥小鼠急性肺損傷具有一定的保護(hù)作用，其機(jī)制可能與調(diào)控 PI3K-Akt、NLRP3/TNF-α等通路抑制炎癥的發(fā)展有關(guān)，為VAC抗炎作用機(jī)制的深入研究提供了依據(jù)。

Objective Based on network pharmacology and animal experiments to explore the protective mechanism of vaccarin (VAC) in acute lung injury of sepsis mice.Methods The potential targets of VAC and the disease targets related to pulmonary sepsis were collected through the Swiss Target Prediction database and the GeneCards database separately. Then, the common targets for VAC (1, 5 mg·kg^-1) and diseases were obtained using Draw Venn Diagram software. The STRING 11.5 database protein-protein interaction was used to construct PPI network and a drug target disease network was constructed using Cytoscape 3.10.0. Finally, Gene Ontology (GO) enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis for key targets were conducted utilizing the Metascape database. A sepsis model was established by intraperitoneal injection of lipopolysaccharide (LPS) and treated with VAC or dexamethasone (DEX). Then, lung tissue and serum were collected from each group, and HE, Masson, and TUNEL staining were used to observe changes in lung tissue morphology, fibrosis area, and cell apoptosis. The levels of inflammatory factors in serum and lung tissue were detected by ELISA and qRT-PCR, respectively. The expression of inflammatory pathway related proteins in lung tissue was detected by immunohistochemistry and Western blotting.Results There were 44 common targets for VAC and pulmonary sepsis. A total of 924 GO items were obtained by GO enrichment analysis, including 823 for biological processes, 52 for cellular composition, and 49 for molecular functions. KEGG analysis identified 20 signaling pathways, including cancer pathway, PI3K-Akt, and JAK-STAT signaling pathway. The animal experiment results showed that compared with the control group, the model group mice had severe lung tissue damage and fibrosis, lung index significantly increased (P < 0.05), and the expression of inflammatory factor in serum and tissue was increased(P < 0.01, 0.001). While, compared with model group, VAC and DEX improved pathological morphology of lung tissue, reduced fibrosis and the expression levels of serum TNF-α, IL-1β, IL-6（P < 0.05, 0.01, 0.001). The expression of inflammatory proteins in lung tissue, and the number of cell apoptosis decreased, while the expression of PI3K and AKT1 proteins increased（P < 0.05, 0.01, 0.001). The results of animal experiments were consistent with the results of network pharmacology.Conclusion VAC have a certain protective effect on acute lung injury in sepsis mice, and its mechanism may be related to the regulation of PI3K-Akt and NLRP3/TNF- α pathways related to its occurrence and development, which provides a basis for the in-depth study of the anti-inflammation mechanism of VAC.

R285.5

國家自然科學(xué)基金面上項(xiàng)目（82170424）；國家自然科學(xué)基金青年項(xiàng)目（81700364）；江蘇省大學(xué)生創(chuàng)新創(chuàng)業(yè)訓(xùn)練計(jì)劃重點(diǎn)項(xiàng)目（202210295035Z）；江南大學(xué)實(shí)驗(yàn)室管理專項(xiàng)研究課題（JDSYS201924）