上调自噬对幽门螺杆菌感染后人胃上皮细胞GES-1的保护作用  

Protective effect of autophagy upregulation on Helicobacter pylori-infected human gastric epithelium cell(GES-1 )

作　　者：熊励晶 童煜[2] 谢晓丽 毛萌[1,2] 

机构地区：[1]四川省成都市妇女儿童中心医院儿童消化内科,610091 [2]四川大学华西第二医院发育与妇儿疾病四川省重点实验室,成都610041

出　　处：《中华消化病与影像杂志（电子版）》2015年第3期35-38,共4页Chinese Journal of Digestion and Medical Imageology(Electronic Edition)

摘　　要：目的观察幽门螺杆菌感染人胃上皮细胞后自噬的发生及变化情况,并探索上调幽门螺杆菌感染后人胃上皮细胞GES-1自噬反应对细胞是否具有一定的保护作用,为进一步研究幽门螺杆菌感染与细胞自噬奠定基础。方法使用一定浓度的幽门螺杆菌感染人胃上皮细胞GES-1,通过Western blot检测感染不同时间点GES-1中微管相关蛋白轻链3Ⅰ/Ⅱ(LC3Ⅰ/Ⅱ)的转换与Beclin 1的表达,免疫荧光染色观察自噬体的出现。确定Xestospongin B处理人胃上皮细胞GES-1的浓度后,将培养细胞分为三组:幽门螺杆菌感染组、幽门螺杆菌感染+Xestospongin B组、正常培养组。首先确定诱导剂对感染后自噬的上调作用,然后使用微毛细管式细胞分析仪检测细胞凋亡和活性,比较三组细胞存活以及凋亡的差异。结果在GES-1细胞被感染后2 h即可观察到自噬的发生,且呈时间依赖性,于8 h到达高峰,24 h开始恢复基础水平。加诱导后细胞自噬水平上调。单纯感染组细胞活性先迅速降低,2 h之后细胞活性平稳降低,凋亡率逐渐增加。加诱导剂组细胞活性在感染后的6 h内细胞活性稍有降低,6 h后细胞活性开始增加,凋亡率降低。结论幽门螺杆菌能够诱导人胃上皮GES-1发生自噬,Xestospongin B能够上调幽门螺杆菌感染后人胃上皮细胞GES-1的自噬水平,并且上调自噬反应能够对幽门螺杆菌感染的人胃上皮细胞GES-1起到一定保护作用。Objective To observe the autophagy induced by Helicobacter pylori （ H.pylori ） infection,and to investigate the protective role of upregulating autophagy of human gastric epithelium cell GES-1 induced by H.pylori infection, and to lay the foundation for the study on H.pylori infection and autophagy.Methods Infected human gastric epithelium cell GES-1 by a certain concentration of H.pylori. Western blot was utilized for detecting the level of LC3-I/II conversion and the level of Beclin 1, which indicated the autophagy induction.The formations of autophagosomes were observed via immunofluorescence with microscopy.GES-1s were divided into three groups：infection group,infection＋Xestospongin B group and control group.Microcapillary cell analyzer （ Millipore Guava easyCyte ） was utilized to measure the cell viability and apoptosis rate,and the differences among the three groups were analyzed.Results At 2 h post infection,autophagy induction became detectable.It reached a peak level at 8 h p.i,while it returned to the basic level at 24 h p.i.Autophagy activities of infected cells were upregulated with induction of Xestospongin B.Cell viability of infection group rapidly decreased firstly,and steadily decreased after 2 hours,and the rate of apotosis was increasing with duration of infection.Cell viability of infection ＋Xestospongin B group decreased sightly within 6 hours post infection and increased after that untill 24 hours,and the rate of apotosis decreased.Conclusions H.pylori is capable of inducing autophagy in human gastric epithelium cell GES-1, which can be modulated by autophagic regulators Xestospongin B.The upregulated autophagy may play a role in protecting GES-1 from H.pylori infection.

关 键 词：螺杆菌 幽门 上皮细胞 自噬 

分 类 号：R57[医药卫生—消化系统]