IκBα突变型基因对缺氧缺糖损伤永生化神经前体细胞的保护作用  

Protective effect of mutated IκBα against damage of immortalized neural progenitor cells induced by oxygen-glucose deprivation

作　　者：祝畅[1] 刘志恒[2] 张传汉[1] 桂伶俐[1] 姚文龙[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院麻醉学教研室,湖北武汉430030 [2]深圳市第二人民医院麻醉科,广东深圳518035

出　　处：《第四军医大学学报》2008年第7期596-598,共3页Journal of the Fourth Military Medical University

基　　金：国家自然科学基金(30300328)

摘　　要：目的:检测在缺氧缺糖环境中,IκBα突变型基因对永生化神经前体细胞株(INPCs)NFκ-B的活性、细胞存活及细胞损伤的影响.方法:在建立转染pcDNA3.1及转染pcD-NA3.1/IκBαM INPCs的基础上,用MTT法检测缺氧缺糖处理3,6和9 h后两细胞株的存活率,用荧光素酶报告基因检测两细胞株缺氧缺糖6 h前后NFκ-B的活性,并测定缺氧缺糖6 h处理后培养液中乳酸脱氢酶的含量,用Hoechst33342染色观察细胞核的形态.结果:转染pcDNA3.1/κIBαM的INPCs中NFκ-B的活性均低于转染pcDNA3.1的INPCs,缺氧缺糖6 h或9 h时,前者存活率高于后者,且6 h时前者上清中的乳酸脱氢酶释放量较少,缺氧缺糖可使两细胞株的部分细胞核呈现凋亡改变.结论:κIBα突变型基因可降低INPCs中NFκ-B的活性,提高INPCs在缺氧缺糖处理后的存活率,减轻细胞损伤.AIM： To determine the effect of mutated IκBα gene on immortalized neural progenitor cell strains （INPCs） in oxygen- glucose deprivation. METHODS： After the establishment of mutated IκBα gene transfected INPCs and blank plasmid transfected INPCs, the cells were exposed to anoxic/hypoglycemic condition for 3, 6 or 9 h respectively. MTr staining was used to detect the survival rate of the two cell strains. Luciferase assay system was used to detect the activity of NF-κB in pcDNA3. 1 transfected or pcDNA3. 1/IκBαM transfected INPCs. Lactate dehydrogenase in culture medium was measured after the cells were exposed to anoxic/hypoglycemic condition for 6 h. The morphological change of two cell strains was observed by Hoechst33342 staining. RESULTS： The activity of NF-κB was down-regulated in pcDNA3.1/IκBαM transfected INPCs before or after oxygen-glucose deprivation. The cell survival rate of pcDNA3.1/ IκBαM transfected INPCs was higher than that of pcDNA3.1 transfected INPCs after exposed to anoxic/hypoglycemic condition for 6 h or 9 h, but there was no difference in cell survival rate at 3 h. After the cells were exposed to anoxic/hypo- glycemic condition for 6 h, lactate dehydrogenase content in culture medium was lower in pcDNA3.1/IκBαM transfected INPCs. Hoechst33342 staining revealed that cells were apoptotic after oxygen-glucose deprivation. CONCLUSION： By down-regulating the activity of NF-κB, mutated IκBα gene improves the cell survival rate of INPCs and lessens the cell damage caused by oxygenglucose deprivation.

关 键 词：干细胞 NF-κB抑制因子α 细胞缺氧 

分 类 号：R743.31[医药卫生—神经病学与精神病学] Q256[医药卫生—临床医学]