ILK对幽门螺杆菌诱导胃上皮细胞GES-1凋亡的影响  

The effect of ILK on apoptosis of GES-1 gastric epithelial cells induced by Helicobacter pylori

作　　者：徐正 丁雲飞 荣倩玉 吴玉龙[1] 赵慧琳[1] 季晓飞[1] 张莹[1] 李波清[1] XU Zheng;DING Yun-fei;RONG Qian-yu;ZHANG Ying;ZHAO Hui lin;JI Xiao-fei;LI Bo-qing(Department of Pathogen Biology,Binzhou Medical University,Yantai 264003,Shandong,China)

机构地区：[1]滨州医学院病原生物学教研室,山东烟台264003

出　　处：《中国病原生物学杂志》2018年第8期819-822,826,共5页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.81771709;No.81471561;No.81702054;No.81501718);滨州医学院青年骨干教师培养计划经费资助项目

摘　　要：目的探讨ILK在幽门螺杆菌诱导胃上皮细胞GES-1凋亡中的作用。方法体外培养GES-1细胞,随机分为空白对照组、Cpd22组和Hp组。采用罗丹明-鬼笔环肽检测不同浓度ILK抑制剂Cpd22对胃上皮细胞形态及细胞骨架改变的影响;Cpd22作用于GES-1细胞1h后,Western blot检测细胞内ILK表达情况;Hp与Cpd22作用1h的GES-1细胞共培养(MOI=100∶1)24h后,采用流式细胞术检测细胞凋亡率,Western blot检测凋亡相关蛋白Caspase-3、PARP表达情况。结果与空白对照组相比,Cpd22浓度为2μmol/L时细胞形态变圆,间隙变大,细胞外缘伪足减少,细胞无法正常伸展。空白对照组与Cpd22组ILK相对表达量分别为0.83±0.01、0.74±0.01,比较差异有统计学意义(t=5.65,P<0.05)。空白对照组、Cpd22组、Hp组细胞凋亡率分别为(6.53±2.78)%、(21.13±5.60)%、(14.07±3.97)%,比较差异有统计学意义(t值分别为-5.52和-6.83,P<0.05);Caspase-3相对表达量分别为0.63±0.07、0.95±0.14、0.88±0.11,差异均有统计学意义(t值分别为-5.44和-5.88,P<0.05);PARP的相对表达量分别为1.17±0.07、0.93±0.16、1.09±0.12,差异有统计学意义(t值分别为3.32和3.46,P<0.05)。Cpd22与Hp共作用组细胞凋亡率为(30.37±5.17)%,Caspase-3的相对表达量为1.07±0.10,PARP的相对表达量为0.86±0.07,与Cpd22组比较差异均有统计学意义(t值分别为4.90、2.85和6.70,P<0.05)。结论抑制ILK表达可增强Hp诱导GES-1细胞凋亡的作用,其机制可能与细胞骨架重排和Caspase-3的表达有关。Objective To examine the role of ILK in the apoptosis of GES-1 gastric epithelial cells induced by Helicobacter pylori. Methods GES-1 cells were cultured in vitro and randomly divided into a blank control group,cells treated with Cpd22,and cells co-cultured with H.pylori.The effects of different concentrations of the ILK inhibitor Cpd22 on the morphology and cytoskeleton of gastric epithelial cells were detected using rhodamine phalloidin.GES-1 cells were treated with Cpd22 for 1 h,and the expression of ILK in cells was detected using Western blotting.GES-1 cells were cocultured with H.pylori（MOI=100：1）and treated with Cpd22 for 1 h.Twenty-four h later,the rate of apoptosis was detected using flow cytometry,and expression of the apoptosis-related proteins Caspase-3 and PARP was detected using Western blotting. Results Compared to the blank control group,when the concentration of Cpd22 was 2μmol/L,the cell morphology was rounder,gaps grew,there were fewer pseudopods on the outer edge of cells,and the cells did not spread normally.The relative expression of ILK was 0.83±0.01 in the blank control group and 0.74±0.01 in cells treated with Cpd22.The level of that relative expression differed significantly in the 2 groups（t=5.65,P〈0.05）.The rate of apoptosis was（6.53±2.78）%in the blank control group,（21.13±5.60）%in cells treated with Cpd22,and（14.07±3.97）%in cells co-cultured with H.pylori.The rate of apoptosis differed significantly among the groups（t=-5.52,-6.83,P〈0.05）.The relative expression of Caspase-3 was 0.63±0.07 in the blank control group,0.95±0.14 in cells treated with Cpd22,and 0.88±0.11 in cells co-cultured with H.pylori.The level of that relative expression differed significantly among the groups（t=-5.44 and-5.88;P〈0.05）.The relative expression of PARP was 1.17±0.07 in the blank control group,0.93±0.16 in cells treated with Cpd22,and 1.09±0.12 in cells co-cultured with H.pylori.The level of that relative expression differed significantly among the groups（t

关 键 词：幽门螺杆菌 细胞凋亡 GES-1细胞 ILK Cpd22 

分 类 号：R378.91[医药卫生—病原生物学]