基于网络药理学及实验验证探讨加味当归芍药散治疗慢性萎缩性胃炎的机制

Mechanism of Modified Danggui Shaoyao San in treatment of chronic atrophic gastritis based on network pharmacology and experimental verification

目的基于网络药理学对加味当归芍药散(modified danggui shaoyao san,MDSS)治疗慢性萎缩性胃炎(chronic atrophic gastritis,CAG)的作用靶点进行预测和动物实验验证。方法利用TCMSP、Swiss Target Prediction、Genecards、OMIM数据库筛选MDSS的活性成分及治疗CAG的作用靶点,利用STRING数据库、Cytoscape3.8软件构建蛋白互作网络并筛选核心靶点,利用Metascape数据库进行GO分析和KEGG通路富集,利用分子对接进行靶点验证。采用N-甲基-N′-硝基-N-亚硝基胍自由饮结合水杨酸钠灌胃制备CAG大鼠模型,采用苏木精-伊红染色法观察大鼠胃黏膜病理情况,透射电镜观察上皮细胞超微结构,采用酶联免疫吸附测定法检测大鼠血清IL-6、IL-10含量,实时荧光定量PCR法、蛋白免疫印迹法检测大鼠JAK2、STAT3、p-STAT3、c-MYC mRNA和蛋白的表达。结果MDSS治疗CAG作用于189个靶点,作用靶点的生物功能与氧化应激反应、细胞凋亡信号传导途径相关,KEGG通路与癌症中的路径JAK-STAT信号传导途径等通路相关。实验结果表明,MDSS能改善CAG大鼠胃黏膜萎缩程度,改善上皮细胞状态,下调CAG大鼠血清IL-6含量,上调IL-10含量,降低胃黏膜JAK2、STAT3、p-STAT3、c-MYC mRNA和蛋白的表达。结论MDSS通过多种活性成分、多靶点与多通路协同作用治疗CAG,其机制可能与抑制JAK2/STAT3信号通路有关。

Aim To predict the targets of Modified Danggui Shaoyao San(MDSS)in the treatment of chronic atrophic gastritis(CAG)based on network pharmacology and vertify the results based on experimention.Methods TCMSP,SWISS TARGETS,GENE CARDS and OMIM databases were used to screen the therapeutic targets of MDSS for CAG.STRING database and Cytoscape software were used to construct the protein interaction network and screen the core targets.Metascape database was used for GO analysis and KEGG enrichment pathways.And molecular docking was used for target validation.CAG rat model was prepared by N-methyl-N′-nitroso-N-nitroguanidine free drink combined with sodium salicylate gastric lavage.The pathology of rat gastric mucosa was observed by hematoxylin-eosin staining,and the ultrastructure of epithelial cells was observed by transmission electron microscopy.The serum IL-6 and IL-10 content was detected by enzyme-linked immunosorbent assay,and the expression of JAK2,STAT3,p-STAT3,c-MYC mRNA and protein in rats was detected by qPCR and Western blot.Results MDSS acted on 189 targets,mainly involved in response to oxidative stress and apoptotic signaling pathway.KEGG analysis related to pathways in cancer and JAK-STAT signaling pathway.The experimental results showed that the MDSS could improve the degree of atrophy of gastric mucosa in CAG rats and improve the status of epithelial cells,down-regulate the serum IL-6 content of CAG rats,up-regulate the IL-10 content,and reduce the expression of JAK2,STAT3,p-STAT3,c-MYC mRNA and protein in gastric mucosa with statistical significance.Conclusions MDSS treats CAG through multiple active ingredients,targets,and pathways,the mechanism of which may be related to the inhibition of the JAK2/STAT3 signaling pathway.