基于网络药理学和实验验证研究柴胡疏肝散治疗慢性萎缩性胃炎的作用机制

Study on the Mechanism of Chaihushugan Powder in Chronic Atrophic Gastritis Based on Network Pharmacology and Experimental Verification

目的通过网络药理学的方法进行预测,再深一步进行动物实验验证来研究柴胡疏肝散治疗CAG的作用机制。方法首先在TCMSP数据库中检索柴胡疏肝散的所有活性成分与药物靶点;通过收集PharmGkb、OMIM、GeneCards和DrugBank数据库中收录的慢性萎缩性胃炎的相关靶点。将药物靶点与疾病靶点进行映射筛选出交集靶点,将得到的交集靶点构建PPI网络与活性成分-共同靶点网络,并对其进行GO和KEGG富集分析。最后利用Vina软件进行分子对接实验验证,并通过免疫印迹法验证柴胡疏肝散对两种受体蛋白EGFR和STAT1的影响。结果最终筛选得到柴胡疏肝散活性成分104个,潜在靶点238个,与慢性萎缩性胃炎的交集靶点52个;GO与KEGG富集分析分别得到2166条目和148条目,主要涉及到JAKSTAT信号通路、TNF信号通路、HIF-1信号通路等;分子对接结果显示EGFR、STAT1两个靶点能够与核心活性成分能够自发结合成较为稳定的构像;免疫印迹法实验证明柴胡疏肝散能够降低大鼠胃黏膜组织EGFR和STAT1蛋白表达。结论通过网络药理学和实验验证,发现柴胡疏肝散可能通过调节EGFR和STAT1蛋白表达来共同调控胃黏膜细胞增殖与凋亡,进而发挥着治疗慢性萎缩性胃炎的效果,为深入进行柴胡疏肝散治疗慢性萎缩性胃炎的作用机制研究提供新思路和新方法。

Objective To study the mechanism of Chaihushugan Powder in the treatment of CAG by using the method of network pharmacology and further animal experiments.Methods Firstly, all the active ingredients and drug targets ofChaihushugan Powder were searched in TCMSP database. Targets for chronic atrophic gastritis were collected fromPharm Gkb, OMIM, Gene Cards, and Drug Bank data. The intersection target was screened by mapping the drug target andthe disease target. The resulting intersection targets were used to construct protein interaction networks and activecomponent-common target networks. GO and KEGG enrichment analysis were performed. Finally, Vina software wasused to verify the molecular docking experiment, and western blotting was used to verify the effect of ChaihushuganPowder on two receptor proteins, EGFR and STAT1. Results 104 active components, 238 potential targets and 52 intersection targets with chronic atrophic gastritis were selected. GO and KEGG enrichment analysis obtained 2166 items and 148 items, respectively, which mainly involved JAK-STAT signaling pathway, TNF signaling pathway andHIF-1 signaling pathway. Molecular docking results showed that EGFR and STAT1 could spontaneously bind to the coreactive components and form a relatively stable conformation by intermolecular forces such as hydrogen bonds. Westernblotting showed that Chaihushugan Powder could reduce the expression of EGFR and STAT1 protein in rat gastricmucosa.Conclusion Through network pharmacology and experimental verification, we find that Chaihushugan Powdermay adjust the EGFR and STAT1 protein expression in regulation of gastric mucosa cell proliferation and apoptosis, andthen play the effect of the treatment of chronic atrophic gastritis, so as to provide new ideas and new methods for in-depth study on the mechanism of action of Chaihushugan Powder in the treatment of chronic atrophic gastritis.