基于网络药理学及实验验证探讨香连化浊方治疗慢性萎缩性胃炎大鼠作用机制  

Mechanism of Xianglian Huazhuo Recipe(香连化浊方)in the Treatment of Chronic Atrophic Gastritis in Rats Based on Network Pharmacology and Experimental Verification

作　　者：刘晓萌 刘建平 郎晓猛 焦红 王玉婷 崔泽华 董紫薇 Liu Xiaomeng;Liu Jianping;Lang Xiaomeng;Jiao Hong;Wang Yuting;Cui Zehua;Dong Ziwei(Graduate School,Hebei University of Chinese Medicine,Shijiazhuang 050091;The First Affiliated Hospital of Hebei University of Chinese Medicine,Shijiazhuang 050011;Key Laboratory of Integrated Chinese and Western Medicine for Gastroenterology Research,Shijiazhuang 050011)

机构地区：[1]河北中医学院研究生院,石家庄050091 [2]河北中医学院第一附属医院,石家庄050011 [3]河北省中西医结合胃肠病研究重点实验室,石家庄050011

出　　处：《中药药理与临床》2022年第4期21-28,共8页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家中医药管理局慢性胃炎(浊毒证)重点研究室项目(编号:200995);国家中医药管理局第三界国医大师传承工作室及全国名老中医传承工作室建设项目(编号:人社部发[2017]45号);第二批国家中医临床研究基地重点病种项目(编号:2018131);国家中医临床研究基地建设项目(编号:国中医药办科技函[2018]18号)。

摘　　要：目的:基于网络药理学和实验验证,探讨香连化浊方治疗慢性萎缩性胃炎(CAG)的作用机制。方法:利用中药系统药理学技术平台(TCMSP)、SWISS TARGETS数据库、基因组注释数据库平台(GENE CARDS)、在线人类孟德尔遗传数据库(OMIM)筛选香连化浊方和疾病的作用靶点,利用STRING数据库、Cytoscape软件构建蛋白互作网络,筛选核心靶点,利用Metascape数据库进行基因本体论(GO)分析与京都基因与基因组百科全书(KEGG)通路富集分析。制备CAG大鼠模型后予香连化浊方干预治疗,观察大鼠胃黏膜组织病理学改变,采用免疫组化、实时荧光定量聚合酶链式反应(qPCR)对核心靶点进行验证。结果:网络药理学结果显示香连化浊方中口服利用度较高的成分有珊瑚甙、过氧化苯甲酰内酯、芍药花酮等;主要活性成分槲皮素、木犀草素、山柰酚等作用于192个靶点,生物学途径主要为参与蛋白磷酸化正调控、氧化应激反应、损伤应答、凋亡信号通路等;细胞组分主要包括膜侧、膜筏、细胞外基质、细胞质的核周围区域等;分子功能包括激酶结合、转录因子结合、蛋白域特异性结合、蛋白激酶活性等。KEGG富集通路与磷脂酰肌醇-3-激酶-丝氨酸苏氨酸蛋白激酶(PI3K-AKT)信号通路、肿瘤坏死因子信号通路、细胞凋亡通路。实验验证研究发现香连化浊方治疗后大鼠胃黏膜萎缩病理程度减轻,胃组织核心靶点AKT1、C-MYC的蛋白表达明显下调(P<0.05),胃组织Cmyc mRNA表达明显下调(P<0.05)。结论:香连化浊方中有效成分槲皮素、木犀草素、山柰酚等可能通过下调Cmyc mRNA、AKT1、C-MYC蛋白表达抑制胃黏膜细胞过度增殖实现对CAG的治疗作用。Objective:To explore the mechanism of Xianglian Huazhuo recipe(香连化浊方)in the treatment of chronic atrophic gastritis(CAG)based on network pharmacology and experimental verification.Methods:The therapeutic targets of Xianglian Huazhuo recipe for CAG were screened out from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),SWISS TARGETS,GENE CARDS,and Online Mendelian Inheritance in Man(OMIM).STRING and Cytoscape were used to construct the protein-protein interaction network and screen out the core targets.Metascape was employed for the gene ontology(GO)annotation and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment of the core targets.The CAG rat model was established by the N-methyl-N’-nitro-N-nitrosoguanidine(MNNG)-based composite modeling method and treated with Xianglian Huazhuo recipe.The histopathological changes of gastric mucosa were observed via hematoxylin-eosin(HE)staining,and the core targets were verified by immunohistochemistry and qPCR.Results:The results of network pharmacology showed that aucubin,Benzoyl lactone peroxide and Paeony flavonoids were the components with high oral availability in Xianglianhuazhuo recipe,and the main active components in Xianglian Huazhuo recipe included quercetin,luteolin,and kaempferol,which acted on 192 targets.The involved biological processes included positive regulation of phosphorylation,oxidative stress response,damage response,and apoptosis,the involved cell components mainly included lateral memebrane,membrane raft,extracellular matrix,and perinuclear region of cytoplasm,and the involved molecular functions included binding to kinase,binding to transcription factor,specific binding to protein domain,and protein kinase activity.The KEGG pathway enrichment analysis showed that the core targets mainly participated in the phosphatidylinositol-3-kinase(PI3 K)-protein kinase B(AKT)signaling pathway,tumor necrosis factor signaling pathway,and apoptosis pathway.The administration of Xianglian Huazhuo

关 键 词：香连化浊方 网络药理学 慢性萎缩性胃炎 丝氨酸/苏氨酸蛋白激酶 C-MYC 

分 类 号：R285.5[医药卫生—中药学]