NF—κB在模拟缺血／再灌注诱导人离体胃黏膜上皮细胞凋亡中的作用  

The role of NF - κB in cultured human gastric mucosal epithelial cell apoptosis induced by simulating ischemia/reperfusion

作　　者：石明[1] 王光明[1] 马鸿基 宋成洁[1] 乔伟丽[1] 

机构地区：[1]徐州医学院生理学教研室,江苏徐州221002

出　　处：《徐州医学院学报》2013年第3期155-158,共4页Acta Academiae Medicinae Xuzhou

基　　金：徐州医学院院课题（2010KJ14）

摘　　要：目的观察核因子KB（NF—κB）在模拟缺血／再灌注诱导人离体胃黏膜上皮细胞凋亡中活性的变化，并进一步阐明其作用。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）＋兀糖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，用流式细胞术、免疫组化法、免疫印迹法观察正常对照组、模拟缺血／再灌注损伤组（I—R组）、NF—κB的抑制剂PDTC预处理组（使用10μmol·L^-1PDTC预处删胃黏膜上皮细胞24h）于模拟缺血1h、再灌注6h后细胞凋亡及NF—κB的表达情况。结果①I—R组细胞凋亡率较正常对照组明显增高（P〈0．01），PDTC预处理组细胞凋亡率较I—R组明显下降（P〈0．01）。②正常对照组胃黏膜上皮细胞核内极少有NF—κB表达，I—R组细胞核内NF—κB表达明显增加，PDTC预处理绀NF—κB核转化现象减轻，胞核染色明显减少。③I—R组与正常对照组比较，胃黏膜上皮细胞核NF—κB蛋白表达水平显著增高（P〈0．01）；PDTC预处理组与I—R组比较，NF—κB蛋白表达水平明显降低（P〈0．01）。结论模拟缺血／再灌注能增加人肖黏膜上皮细胞核内NF—κB的表达，NF—κB的激活具有促进细胞凋产的作用；PDTC叮以抑制细胞凋亡，对人胃黏膜上皮细胞模拟缺血／再灌注损伤有保护作用。Objective To investigate the effect of NF - κB in the cuhured gastric mucosa epithelial cell on ischemi- a/repeffusion induced apoptosis. Methods The epithelial cells of gastric mucosa were exposed to hypoxia（5% CO2, 1% 0=, 94% N2 ） in glucose - free KR bicarbonate bufier and succedent reoxygenation （5% CO=, 95% air） in DMEM/ F12 to simulate conditions of ischemia and repeffusion. PDTC, a NF - KB inhibitor was pretreated with cells for 24 h be- fore simulating ischemia and reperfusion. The apoptotic cells were evaluated by flow cytometry （FCM） afte 1 h simulated ischemia and 6 h repeffusion. Immunohistochemical staining and immunoblotting analysis expression of NF -κB in cul- tured gastric mucosa epithelial cells. Results （1）FCM analysis revealed that the percentage of cell apoptosis increased significantly after simulated ischemia and repeffusion compared with the control group （P 〈 0.01 ）. However, PDTC re- duced the percentage of apoptotic cells. （2） [mmunohistochemical staining showed that expression of NF - κB in nucleus increased and PDTC inhibited the NF - κB translocated into nucleus, showing less NF - κB staining compared with simu- lated ischemia and repeffusion. （3） Stfinulated ischemia and repeffusion increased the NF - κB protein expression while PDTC inhibited the effect. Conclusion The NF - κB can be activated after simulated ischemia/repeffusion injury. There is protective effect of specific NF - κB inhibitor PDTC on apoptosis induced by ischemia/repeffusion injury.

关 键 词：胃缺血 阿灌注 胃黏膜上皮细胞 细胞凋亡 核因子κB 硫代氨基甲酸吡咯烷 

分 类 号：R573[医药卫生—消化系统]