目的 通过数据库预测和分子对接技术探究小儿广朴止泻口服液治疗腹泻的活性成分和作用机制，并在大鼠小肠上皮IEC-6细胞上进行初步实验验证。方法 通过中药系统药理学平台（TCMSP）数据库和文献检索获取小儿广朴止泻口服液的化学成分；采用GeneCards和DisGeNET数据库获得腹泻相关基因，构建蛋白质-蛋白质相互作用（protein-protein interaction network，PPI）网络获得小儿广朴止泻口服液治疗腹泻的重要靶点，并进行基因本体（gene ontology，GO）功能及京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes，KEGG）通路富集分析；使用Autodock Vina软件进行分子对接验证。采用脂多糖（lipopolysaccharide，LPS）诱导IEC-6细胞损伤模型，采用MTT、ELISA法检测小儿广朴止泻口服液对细胞凋亡及炎症的影响，并通过Western blotting法验证其可能的作用机制。结果 小儿广朴止泻口服液的主要活性成分有63个，预测获得重要靶点86个，核心靶点28个；GO功能和KEGG通路富集分析显示小儿广朴止泻口服液主要调控了机体炎症反应、细胞增殖、细胞凋亡等生物过程，涉及肿瘤坏死因子（tumor necrosis factor，TNF）、Toll样受体（Toll-like receptor，TLR）、缺氧诱导因子-1（hypoxia inducible factor-1，HIF-1）以及磷酸肌醇-3-激酶（phosphatidylinositol-3- kinase，PI3K）-蛋白激酶B（protein kinase B，Akt）等信号通路。体外结果表明小儿广朴止泻口服液能显著升高LPS诱导的IEC-6细胞存活率（P＜0.01），显著抑制炎症因子白细胞介素-6（interleukin-6，IL-6）、IL-1β和TNF-α的分泌（P＜0.01），显著下调剪切型半胱氨酸天冬氨酸蛋白酶-3（cleaved cystein-asparate protease-3，cleaved Caspase-3）、Caspase-8、核因子-κB p65（nuclear factor-κB p65，NF-κB p65）蛋白表达（P＜0.01），上调β-catenin蛋白表达（P＜0.01）。结论 小儿广朴止泻口服液可能通过抑制炎症反应和细胞凋亡治疗腹泻。

Objective To explore the active ingredients and mechanism of Xiaoer Guangpu Zhixie Oral Liquid (小儿广朴止泻口服液) in treatment of diarrhea by database prediction and molecular docking, and preliminarily verify it on rat intestinal epithelium IEC-6 cells. Methods TCMSP database and literature retrieval were applied to obtain chemical components of Xiaoer Guangpu Zhixie Oral Liquid. Disease targets relating to diarrhea were screened out through GeneCards and DisGeNET database. A protein-protein interaction (PPI) network was constructed, and core targets in network were predicted. Gene ontology (GO) function and Kyoto encyclopedia of genes and genomes (KEGG) pathway enrichment analysis of core targets were performed. Furthermore, molecular docking method was used to verify the binding of components with core targets by Autodock Vina. The injury model of IEC-6 cells was induced by lipopolysaccharide (LPS), effects of Xiaoer Guangpu Zhixie Oral Liquid on apoptosis and inflammation were detected by MTT and ELISA, and possible mechanism was verified by Western blotting. Results There were 63 major active compounds in Xiaoer Guangpu Zhixie Oral Liquid, with 86 significant targets and 28 core targets. The results of GO function and KEGG pathway enrichment analysis revealed Xiaoer Guangpu Zhixie Oral Liquid mainly regulated the organism’s inflammatory response, cell proliferation, apoptosis and other biological processes. The mainly involved signal pathways included tumor necrosis factor (TNF) signaling pathway, Toll-like receptor (TLR) signaling pathway, hypoxia inducible factor-1 (HIF-1) signaling pathway and phosphatidylinositol-3-kinase (PI3K)-protein kinase B (Akt) signaling pathway. In vitro experiment confirmed that Xiaoer Guangpu Zhixie Oral Liquid could significantly increase the survival rate of IEC-6 cells induced by LPS (P < 0.01), inhibit the release of inflammatory factors IL-6, IL-1β and TNF-α (P < 0.01), down-regulate cleaved cystein-asparate protease-3 (cleaved Caspase-3), Caspase-8 and nuclear factor-κB p65 (NF-κB p65) protein expressions (P < 0.01), and up-regulate β-catenin protein expression (P < 0.01).Conclusion Xiaoer Guangpu Zhixie Oral Liquid may treat diarrhea by inhibiting inflammatory response and apoptosis.

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