重组腺病毒介导的Bcl-2基因转染对缺氧再复氧肠上皮细胞的保护作用  

Protective effects of adenovirus-mediated Bcl-2 gene transfection for the intestinal epithelial cells against hypoxia-reoxygenation in vitro

作　　者：伊远学[1,2,3] 刘筑 罗效梅 刘晓平[1,2] 李宝金[1,2] 张超[1,2] 冷希圣[1,2] 

机构地区：[1]北京大学香港科技大学医学中心 [2]北京大学深圳医院肝胆腔镜外科,518036 [3]武警重庆总队医院检验科,400061

出　　处：《检验医学与临床》2010年第17期1796-1798,共3页Laboratory Medicine and Clinic

基　　金：广东省医学科研基金(A2007595);广东省自然科学基金博士启动项目(06300934);国家自然科学基金资助项目(30371386)

摘　　要：目的探讨复制缺陷型重组腺病毒介导的Bcl-2基因转染对缺氧再复氧损伤肠上皮细胞的保护作用。方法构建能介导Bcl-2基因转染和表达的复制缺陷型重组腺病毒载体(Ad/CMV-Bcl-2),转染体外培养的小肠上皮细胞株(IEC-6),检测其Bcl-2基因表达变化,分别对正常对照组、缺氧复氧组(单纯给予缺氧复氧处理)、Ad-CMV/Bcl-2转染组(AdCMV/Bcl-2腺病毒载体转染48h后再给予缺氧复氧处理)细胞的凋亡率、死亡率及活力进行分析。结果对照组微弱表达,Ad/CMV-Bcl-2转染组细胞Bcl-2基因高表达;经缺氧再复氧处理后,Annexin-V-Flous试剂盒检测死亡细胞明显减少(P<0.01),凋亡显著受到抑制(P<0.01);采用MTT法证明,Ad/CMV-Bcl-2转染组细胞活力较对照组明显增强(P<0.01)。结论复制缺陷型重组腺病毒介导的Bcl-2基因转染可显著减轻缺氧再复氧导致的肠上皮细胞损伤。Objective To investigate the possibility of protection of intestinal epithelial cells against hypoxia-reoxygenation through recombinant adenovirus-mediated Bcl-2 gene transfection.Methods Replication defective recombinant adenoviral vector Ad/CMV-Bcl-2 had been constructed with full length human Bcl-2 gene.The overexpression of Bcl-2 gene was detected by RT-PCR in transfected groups.After the cells suffered 1hour of hypoxia followed by 1 hour of reoxygenation,the effects of transfected Bcl-2 gene overexpression were studied.The cell viability was analyzed by MTT method,and the apoptosis and dead cell ratio were evaluated with Annexin-V-Flous stained kit in vitro.Results The expression of Bcl-2 was positive in the Ad/CMV-Bcl-2 transfected intestinal epithelial cells and negative in the control group.After hypoxia-reoxygenation,the cell viability rates of Bcl-2 gene transfected groups were all significantly higher than that of control group（P〈0.01）.while the apoptosis rates of Bcl-2 gene transfected groups were significantly lower than that of control group （P〈0.01）.Conclusion The experimental results of transfection in vitro showed the indeed protection of the injury intestinal epithelial cells caused by the hypoxia/reoxygenation.

关 键 词：Bcl-2 腺病毒 转染 细胞低氧 

分 类 号：R341[医药卫生—基础医学]