摘要
目的:通过网络药理学方法及体内实验研究探讨霍山石斛治疗胃溃疡(GU)的作用机制。方法:从中药系统药理学数据库与分析平台(TCMSP)数据库及文献中收集建立霍山石斛的成分库,寻找其主要活性化合物,并通过TCMSP等数据库预测作用靶点。从GeneCards、在线人类孟德尔遗传数据库(OMIM)、DisGeNET等数据库中筛选GU相关基因,将成分靶点与疾病基因交集后构建蛋白质-蛋白质相互作用(PPI)网络,并进行基因本体(GO)富集分析和京都基因与基因组百科全书(KEGG)富集分析。根据预测结果采用苏木素-伊红(HE)染色法、免疫组化、酶联免疫吸附测定法(ELISA)、蛋白免疫印迹法(Western blot)、实时荧光定量聚合酶链式反应(Real-time PCR)等实验验证霍山石斛对乙酸诱导GU大鼠模型的影响。结果:筛选出霍山石斛63种有效成分,得到霍山石斛治疗GU的靶基因37个,PPI分析得到多个霍山石斛治疗GU的可能核心靶点,并通过作用于磷脂酰肌醇3-激酶(PI3K)/蛋白激酶B(Akt)、缺氧诱导因子-1(HIF-1)、肿瘤坏死因子(TNF)、核转录因子-κB(NF-κB)等信号通路和各种生物过程影响GU的发展。体内实验结果显示,霍山石斛可显著降低模型大鼠血清中白细胞介素-1β(IL-1β),白细胞介素-6(IL-6)及TNF-α等炎性因子的水平(P<0.05,P<0.01),增加溃疡边缘的胃血流量(GBF),增加溃疡边缘表皮生长因子(EGF),表皮生长因子受体(EGFR)的表达(P<0.01),显著下调GU大鼠胃组织中PI3K,Akt蛋白和mRNA的表达,上调人第10号染色体缺失的磷酸酶(PTEN)蛋白和mRNA的表达(P<0.01)。结论:网络药理学分析及动物实验结果提示,霍山石斛可以通过调节EGFR/PI3K/Akt信号通路抑制组织炎症,增加溃疡边缘的胃微循环血流,促进细胞增殖和受损胃黏膜的修复。
Objective:To explore the mechanism of Dendrobium huoshanense in the treatment of gastric ulcer(GU)based on network pharmacology and in vivo experiment.Method:The active components of D.huoshanense were searched from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and literature,and the targets of the components were screened from TCMSP and SwissTargetPrediction.GU-related genes were retrieved from GeneCards,Online Mendelian Inheritance in Man(OMIM),and DisGeNET.Thereby,the common targets of the disease and the medicinal were yielded and the protein-protein interaction(PPI)network was constructed,followed by Gene Ontology(GO)term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment.According to the predicted results,hematoxylin-eosin(HE)staining,immunohistochemistry,enzyme-linked immunosorbent assay(ELISA),Western blot,and real-time fluorescent quantitative polymerase chain reaction(Real-time PCR)were used to validate the effects of D.huoshanense on acetic acid-induced GU in rats.Result:A total of 63 active components of D.huoshanense and 37 target genes of D.huoshanense for the treatment of GU were screened out.PPI network analysis yielded several possible core anti-GU targets of D.huoshanense.They influenced the development of GU by acting on signaling pathways such as phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt),hypoxia inducible factor-1(HIF-1),tumor necrosis factor(TNF),and nuclear factor-κB(NF-κB),and various biological processes.The in vivo experiment showed that D.huoshanense significantly reduced the levels of inflammatory factors such as interleukin-1β(IL-1β),interleukin-6(IL-6),and TNF-αin the serum of model rats(P<0.05,P<0.01),increased gastric blood flow(GBF)at the ulcer margin,raised the expression of epidermal growth factor(EGF)and epidermal growth factor receptor(EGFR)at the ulcer margin(P<0.01),significantly down-regulated protein and mRNA expression of PI3K and Akt,and up-regulated protein and mRNA expression of phosphatase and tensin homolog deleted on chromosome ten(PTEN)in the gastric tissues of GU rats(P<0.01).Conclusion:Through regulating EGFR/PI3K/Akt signaling pathway,D.huoshanense can inhibit tissue inflammation,increase gastric microcirculatory blood flow at the ulcer margin,and promote cell proliferation and repair of damaged gastric mucosa.
作者
桂文琪
方媛
聊晓玉
孙纪
俞年军
彭代银
韩岚
GUI Wen-qi;FANG Yuan;LIAO Xiao-yu;SUN Ji;YU Nian-jun;PENG Dai-yin;HAN Lan(School of Pharmacy of Anhui University of Chinese Medicine,Anhui Province Key Laboratory of Chinese Medicinal Compounds,Synergetic Innovation Center of Anhui Authentic Chinese Medicine,Heifei 230012,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2022年第7期151-161,共11页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(U19A2009)
安徽高校科研创新平台团队项目(皖教秘科[2015]49)。
