模拟缺血-再灌注诱导人离体胃黏膜上皮细胞的凋亡.docVIP

　　模拟缺血/再灌注诱导人离体胃黏膜上皮细胞的凋亡 作者：石玥，乔伟丽，王光明，吴金霞，闫长栋，张建福 【摘要】 目的 模拟缺血/再灌注诱导人离体的胃黏膜上皮细胞损伤模型，观察再灌注不同时间点胃黏膜上皮细胞凋亡情况并分析其 规律 。方法 采用人胃黏膜上皮细胞株进行体外培养并传代，细胞以缺氧(5% CO2、1% O2、94% N2)+无糖KR液模拟缺血、缺氧，复氧、复糖模拟再灌注诱导胃黏膜上皮细胞凋亡，用流式细胞仪、Hoechst33258染色法观察正常对照组及模拟损伤组于模拟缺血1 h，再灌注3、6、12 h细胞凋亡的情况及变化规律。结果 ①胃黏膜上皮细胞12～24 h贴壁，24 h后有少量上皮样细胞从周边爬出，48 h后细胞呈指数样生长，72 h后细胞已基本能达到融合。②模拟胃缺血/再灌注后细胞凋亡率于缺血1 h、再灌注6 h时出现高峰，且与正常对照组相比差异有统计学意义(Plt;0.01)。经Hoechst33258染色，荧光显微镜下观察可见细胞于缺血1 h、再灌注6 h时产生典型的凋亡形态学变化如核浓染、核碎裂、边集等。结论 成功模拟了缺血/再灌注诱导人离体的胃黏膜上皮细胞损伤模型;该损伤模型可引起胃黏膜上皮细胞的凋亡，且在缺血1 h、再灌注6 h时为高峰。 【关键词】 胃缺血/再灌注;胃黏膜上皮细胞;细胞凋亡;细胞培养 　　Abstract: Objective To simulate the ischemia/reperfusion-induced injury to the epithelial cells of gastric mucosa and to investigate the cellular apoptosis at different time points in order to explore its regularity.Methods Human gastric epithelial cell line ulate conditions of ischemia and hypoxia, reoxygenation and glucose rEintroduction. The morphological changes and the percentage of apoptotic cells etry (FCM) at reperfusive 3, 6 and 12 h secondary to 1 h simulated ischemia culture.Results ①Observation under the inverted microscope revealed the gastric mucosa epithelial cell adherence after 12 to 24 h; and at 24th h, there ulated ischemic 1 h and reperfusive 6 h, icroscopy revealed typical morphological changes of apoptotic cells including hyperchromatic nuclEI, karyorrhexis, margination, etc. at ischemic 1 h and reperfusive 6 h.Conclusion The ischemia/reperfusion-induced apoptosis of human gastric mucosal epithelial cells cultured in vitro ulated. This injury model is able to induce apoptosis of gastric mucosal epithelial cells e point of reperfusive 6th h secondary to ischemic 1st h. 　　Key ia-reperfusion; gastric mucosa epithelial cell; cell apoptosis; cell culture 　　胃缺血/再灌注(gastric ischemia-reperfusion, GI/R)损伤是一种常见的临床病理生理过程，临床发生多器官障碍综合征时，由于血液的重分布造成的胃肠黏膜缺血出现的最早最明显，远比其他脏器更易发生缺血-再灌注损伤，因此，胃被认为是机体内最早受累的器官[1-2]。胃黏膜虽易损伤，但其修复快，它是机体内自身代谢最快的组织之一，这表明胃黏膜细胞可能具有较强的抗凋亡调控机制和增殖修复能力。深入研究GI-R的胃黏膜上皮细胞凋亡、增