CAJ | 학술논문

目的观察自噬在乙醇诱导胃黏膜上皮GES-1细胞凋亡中的作用.方法体外使用100～1 600 mmol/L浓度乙醇诱导胃黏膜上皮GES-1细胞发生凋亡,并分为4组:空白对照组、乙醇组、乙醇+雷帕霉素(RAPA)组和乙醇+氯喹(CQ)组.流式细胞仪检测各组细胞凋亡水平,噻唑蓝(MTT)法检测各组细胞增殖程度,免疫荧光显微镜观察自噬小体,Western blot检测各组中微管相关蛋白1轻链32β(LC3)的表达水平.结果细胞凋亡随着乙醇浓度增加而升高,自噬能有效降低这种凋亡.乙醇组细胞凋亡率[(17.78±7.86)％]明显高于空白对照组[(12.64±6.82)％]和乙醇+RAPA组[(14.88±7.12)％],同时又显著低于乙醇+CQ组[(21.44±8.69)％,P＜0.05].在乙醇干预下,诱导细胞自噬可以增加细胞的相对增殖活性.结论乙醇可以诱导胃黏膜上皮GES-1细胞凋亡.促进细胞自噬可以有效降低乙醇诱导的细胞凋亡,抑制自噬可以有效增高乙醇诱导的细胞凋亡.
목적 관찰자서재을순유도위점막상피GES-1세포조망중적작용.방법 체외사용100～1 600 mmol/L농도을순유도위점막상피GES-1세포발생조망,병분위4조:공백대조조、을순조、을순+뢰파매소(RAPA)조화을순+록규(CQ)조.류식세포의검측각조세포조망수평,새서람(MTT)법검측각조세포증식정도,면역형광현미경관찰자서소체,Western blot검측각조중미관상관단백1경련32β(LC3)적표체수평.결과 세포조망수착을순농도증가이승고,자서능유효강저저충조망.을순조세포조망솔[(17.78±7.86)％]명현고우공백대조조[(12.64±6.82)％]화을순+RAPA조[(14.88±7.12)％],동시우현저저우을순+CQ조[(21.44±8.69)％,P＜0.05].재을순간예하,유도세포자서가이증가세포적상대증식활성.결론 을순가이유도위점막상피GES-1세포조망.촉진세포자서가이유효강저을순유도적세포조망,억제자서가이유효증고을순유도적세포조망.
Objective To investigate the impact of autophagy on the level of apoptosis induced by ethanol in gastric epithelial cells (GES-1) in vitro.Methods GES-1 cells were treated with ethanol at 100-1 600 mmol/L as an ethanol-induced apoptotic model.GES-1 cells were divided into 4 groups:control group,ethanol group,ethanol and rapamycin (RAPA,autophagy revulsant) group,and ethanol and chloroquine (CQ,autophagy inhibitor) group.Apoptosis rate was measured using flow cytometry.The methyl thiazol tetrazolium (MTT) assay was used to evaluate the cell proliferation.Autophagic level was measured by immunofluorescence microscopy.The expression of autophagy related proteins microtubule2associated protein 1 light chain 32β3(LC3) was detected by using Western blotting.Results Induction of autophagy could significantly reduce ethanol-induced apoptosis of GES-1 cells.Apoptosis rate in ethanol group [(17.78 ± 7.86) ％] was higher than that in control group [(12.64 ± 6.82) ％,P ＜ 0.05] and RAPA group [(14.88 ± 7.12) ％,P ＜ 0.05],but lower than the ethanol and chloroquine group [(21.44 ± 8.69) ％,P ＜ 0.05].Additionally,under the intervention of ethanol,up-regulation of autophagy could promote the cell proliferation of GES-1 cells.Conclusion Ethanol may induce the apoptosis of gastric epithelial cells.Up-regulation of autophagy protects GES-1 cells against ethanol-induced apoptosis while inhibition of autophagy may aggravate ethanol-induced apoptosis of GES-1 cells.