幽门螺杆菌感染胃上皮细胞诱导血管生成素样蛋白4表达的机制及其功能研究  

Regulatory and functional mechanism of angiopoietin-like protein-4 in gastric epithelial cells with Helicobacter pylori infection

作　　者：陈婉燕 岳耕羽 徐小林 庄园[1] CHEN Wanyan;YUE Gengyu;XU Xiaolin;ZHUANG Yuan(Department of Microbiology and Biochemical Pharmacy,Faculty of Pharmacy and Laboratory Medicine,Army Medical University(Third Military Medical University),Chongqing,400038;Department of Gastroenterology,Affiliated Hospital of Zunyi Medical University,Zunyi,Guizhou Province,563000,China)

机构地区：[1]陆军军医大学(第三军医大学)药学与检验医学系微生物与生化药学教研室,重庆400038 [2]遵义医科大学附属医院消化内科,贵州遵义563000

出　　处：《陆军军医大学学报》2024年第9期997-1006,共10页Journal of Army Medical University

基　　金：国家自然科学基金面上项目(82070578)。

摘　　要：目的 探究血管生成素样蛋白4(angiopoietin-like 4,ANGPTL4)在幽门螺杆菌(Helicobacter pylori,H.pylori)感染的胃上皮细胞中的表达调控及其功能机制。方法 收集2021年11月陆军军医大学第二附属医院消化内科胃镜室H.pylori阳性患者的胃镜标本以及H.pylori感染小鼠的胃组织,利用免疫荧光染色技术检测ANGPTL4在胃组织中的表达部位;用H.pylori感染胃上皮细胞和胃类器官,运用实时荧光定量PCR和Western blot等检测ANGPTL4的表达;探讨H.pylori感染诱导ANGPTL4表达的调控机制;利用ANGPTL4重组蛋白刺激胃上皮细胞,检测其对密封蛋白1 (claudin 1,CLDN1)表达的影响。结果 H.pylori感染能显著诱导胃上皮细胞ANGPTL4表达增加,且具有毒力因子CagA依赖性、细菌浓度依赖性和感染时间依赖性(P<0.05);相对于未感染组和CagA敲除(△cagA)H.pylori感染组,野生型(wild-type, WT)H.pylori感染组的人和小鼠胃类器官ANGPTL4表达显著上调(P<0.05);调控机制方面,阻断信号通路NF-κB可明显抑制胃上皮细胞ANGPTL4表达增加(P<0.05);功能机制方面,ANGPTL4重组蛋白通过激活通路ERK下调胃上皮细胞CLDN1表达(P<0.05)。结论 H.pylori依赖毒力因子CagA通过激活NF-κB信号通路诱导胃上皮细胞表达ANGPTL4;ANGPTL4激活通路ERK下调胃上皮细胞CLDN1表达,从而影响H.pylori感染相关胃部疾病的发展。Objective To explore the regulatory and functional mechanism of angiopoietin-like 4(ANGPTL4)expression in gastric epithelial cells infected with Helicobacter pylori(H.pylori).Methods H.pylori-positive gastric specimens from Department of Gastroenterology of second Affiliated Hospital of Army Medical University in November 2021 and the gastric tissues of H.pylori-infected mice were collected,and the expression locations of ANGPTL4 in these gastric tissues were detected by immunofluorescence assay.After gastric epithelial cells and gastric organoids were infected with H.pylori,the expression of ANGPTL4 was detected by real-time PCR and Western blotting.The regulatory mechanism of H.pylori-induced ANGPTL4 expression was investigated by cellular models above.Recombinant ANGPTL4 was used to stimulate gastric epithelial cells to investigate the effect of ANGPTL4 on the claudin 1(CLDN1)expression.Results H.pylori infection significantly increased the expression of ANGPTL4 in gastric epithelial cells in a CagA-dependent,bacterial concentration-dependent and infection time-dependent manner(P<0.05).Compared to uninfected cells and CagA knockout(△cagA)H.pylori-induced ones,the ANGPTL4 expression was significantly increased in WT H.pylori-infected human and mouse gastric organoid(P<0.05).Blocking NF-κB signaling pathway significantly inhibited the enhanced ANGPTL4 expression in H.pylori-infected gastric epithelial cells(P<0.05).Recombinant ANGPTL4 activated ERK signaling pathway to down-regulate the CLDN1 expression in gastric epithelial cells(P<0.05).Conclusion H.pylori activates NF-κB signaling pathway to up-regulate ANGPTL4 expression in gastric epithelial cells in a CagA-dependent manner,and ANGPTL4 activates ERK signaling pathway to down-regulate the CLDN1 expression in gastric epithelial cells,which may contributes to the development of gastric diseases induced by H.pylori infection.

关 键 词：幽门螺杆菌 血管生成素样蛋白4 NF-ΚB信号通路 密封蛋白1 

分 类 号：R322.44[医药卫生—人体解剖和组织胚胎学] R378.99[医药卫生—基础医学] R341