基于网络药理学和分子对接结合体内实验探究黄花白及抗胃溃疡的作用机制

Network-based pharmacology combined with in vivo experiments to investigate the mechanism of action of Bletilla ochracea against gastric ulcers

目的基于网络药理学和分子对接结合体内实验方法探究黄花白及抗胃溃疡的作用机制。方法通过文献及TCMSP、TCMIP数据库筛选出黄花白及的活性成分及靶点,在GeneCard数据库中获取GU靶点,取药物和疾病共同靶点构建蛋白互作网络(PPI),得到核心靶点后进行GO分析和KEGG富集分析,并采用分子对接对网络药理学预测结果进行验证。根据预测的结果采用苏木素-伊红(HE)染色法和酶联免疫吸附测定法(ELISA)验证黄花白及对乙酸诱导GU大鼠模型的影响。结果从文献中筛选出8种黄花白及活性成分,删除重复靶点后得到129个潜在作用靶点。从GeneCard数据库中获取GU靶点,取药物靶点与GU靶点交集得到共同靶点70个,构建蛋白互作网络(PPI)得到黄花白及治疗GU核心靶点有34个,这些靶点通过作用于IL-17、NF-κB、TNF和MAPK等介导的信号通路和各种生物过程影响GU的发展。分子对接结果表明,黄花白及主要活性成分与核心靶点IL-6、TNF、IL-1β和NF-κB具有良好的对接活性。动物实验结果表明,黄花白及能改善GU大鼠胃黏膜损伤,并且显著降低胃溃疡大鼠胃组织中IL-6、IL-1β、TNF-α和MPO等炎性因子的水平,增加EGF的水平(P<0.05)。结论黄花白及可以通过调节IL-17、NF-κB、TNF和MAPK等介导的信号通路抑制组织炎症,分子对接证实了以上网络药理学预测的可靠性,可能通过调控这些通路中的IL-6、TNF、IL-1β、EGF和MPO发挥治疗GU的作用。

Objective To investigate the mechanism of action of Bletilla ochracea against gastric ulcer(GU)based on network pharmacology combined with in vivo experimental methods.Methods Screening the active ingredients and targets of Bletilla ochracea through literature and TCMSP and TCMIP databases.Get GU targets in GeneCard database,take drug and disease common targets to construct protein interactions network(PPI),get core targets and then perform GO analysis and KEGG enrichment analysis,and use molecular docking to validate the network pharmacological prediction results.Based on the predicted results,hematoxylin-eosin(HE)staining and enzyme-linked immunosorbent assay(ELISA)were used to verify the effect of Bletilla ochracea on acetic acid-induced GU rat model.Results Eight active ingredients of Bletilla ochracea were screened from the literature,and 129 potential targets were obtained after the deletion of repeated targets.GU targets were obtained from GeneCards.The intersection of drug targets and GU targets yielded 70 targets.Protein interaction network analysis showed 34 core targets of Bletilla ochracea for GU treatment.These targets affect the development of GU by acting on the signaling pathways such as IL-17,NF-κB,TNF and MAPK,and various biological processes.The molecular docking results showed that the main active components of Bletilla ochracea had good docking activities with the core targets IL-6,TNF,IL-1βand NF-κB.The results of animal experiments showed that Bletilla ochracea ameliorated gastric mucosal injury in rats with GU,and significantly reduced the levels of inflammatory factors such as IL-6,IL-1β,TNF-αand MPO,and increased the levels of EGF in gastric tissues of rats with(P<0.05).Conclusion Bletilla ochracea can inhibit tissue inflammation by regulating signaling pathways such as IL-17,NF-κB,TNF and MAPK.Molecular docking confirms the reliability of the above network pharmacology predictions and may exert therapeutic GU effects by modulating IL-6,TNF,IL-1β,EGF and MPO in these pathways.