VPS4B在克罗恩病发病中的作用机制研究  

Study on the role of VPS4B in the pathogenesis of Crohn′s disease

作　　者：刘玉峰[1] 徐伟松[1] 范辉[1] 顾新红 刘肇修[2] 倪润洲[2] 肖明兵[2] LIU Yufeng;XU Weisong;FAN Hui;GU Xinhong;LIU Zhaoxiu;NI Runzhou;XIAO Mingbing(Department of Gastroenterology,Nantong No.2 People's Hospital,Jiangsu Province,Nantong 226002;Department of Gastroenterology,the Affiliated Hospital of Nantong University)

机构地区：[1]江苏省南通市第二人民医院消化内科,南通226002 [2]南通大学附属医院消化内科

出　　处：《南通大学学报（医学版）》2021年第1期11-14,共4页Journal of Nantong University(Medical sciences)

基　　金：国家自然科学基金资助项目(82000497);南通市科技局项目(MS22020005,MS12019020)。

摘　　要：目的:研究空泡蛋白分选4B(vacuolar protein sorting 4B, VPS4B)在克罗恩病(Crohn′s disease, CD)患者肠黏膜中的表达、作用机制及其临床意义。方法:收集南通市第二人民医院2014年1月—2019年12月CD患者肠黏膜及健康人群肠黏膜石蜡标本各10例,采用免疫组化、蛋白质免疫印迹(Western Blot)检测CD及正常人群肠黏膜活检组织中VPS4B的表达及定位。用三硝基苯磺酸(trinitrobenzenesulphonic acid, TNBS)灌肠构建小鼠结肠炎模型,用Western Blot检测VPS4B、凋亡指标active caspase-3、裂解多聚(ADP-核糖)聚合酶[cleaved poly(ADP-ribose) polymerase, cleaved PARP]以及与炎症性肠病发生密切相关的信号通路p38有丝分裂原活化的蛋白激酶(p38 mitogen-activated protein kinase, p38 MAPK)的表达情况,免疫组化检测VPS4B的分布情况,免疫荧光双标检测VPS4B与active caspase-3的共定位情况。结果:(1)免疫组化结果显示,VPS4B在CD患者肠黏膜中的表达明显高于正常对照组,主要定位在肠上皮细胞。(2)在TNBS诱导的结肠炎小鼠模型中,VPS4B和active caspase-3、cleaved PARP、磷酸化的p38(phosphorylated p38, pp38)的表达均升高,且与active caspase-3有共定位。结论 :VPS4B在CD患者的肠黏膜组织中表达升高,通过调控p38MAPK信号通路促进肠上皮细胞凋亡,可能是CD的重要病理机制之一。Objective:To investigate the expression of vacuolar protein sorting 4 B(VPS4 B)in the intestinal tissues of Crohn′s disease(CD),and further to study its molecular mechanism and clinical significance.Methods:From January 2014 to December2019,at Nantong No.2 People′s Hospital,10 paraffin specimens of intestinal mucosa from Crohn′s disease patients and healthy people were collected.Western Blot and immunohistochemistry were employed to detect the expression and distribution of VPS4 B in mucosal tissues by biopsy from patients with CD and normal controls.A murine model of colitis induced by trinitrobenzenesulphonic acid(TNBS)was established.Western Blot was performed to investigate the expression levels of VPS4 B,apoptotic markers active caspase-3,cleaved poly(ADP-ribose)polymerase(cleaved PARP)and p38 mitogen-activated protein kinase(p38 MAPK).Immunohistochemistry was performed to explore the distribution of VPS4 B.Double-immunofluorescent was performed to detect the co-localization of VPS4 B/active capase-3.Results:(1)Intestinal expression of VPS4 B was increased in patients with CD compared with normal controls and it was mainly located in intestinal epithelial cells.(2)In TNBS-induced colitis,up-regulation of VPS4 B was accompanied with the elevated levels active caspase-3,cleaved PARP and phosphorylated p38(p-p38)in colitis intestinal epithelial cells.VPS4 B was colocalized with active caspase-3 in intestinal epithelial cells.Conclusion:Increased levels of VPS4 B promoted the caspase-mediated intestinal epithelial cells apoptosis through p38 MAPK pathway in CD,which might involved in the pathophysiology of CD.

关 键 词：克罗恩病 空泡蛋白分选4B 磷酸化的p38 小鼠 

分 类 号：R574.5[医药卫生—消化系统]