E1A激活基因阻遏子（CREG）对过氧化氢诱导骨髓间充质干细胞凋亡保护作用的实验研究  

The Experimental research of cellular repressor of E1A-stimulated genes （CREG） inhibiting apoptosis induced by hydrogen peroxide in bone marrow mesenchymal cells

作　　者：张天资[1] 王昕[1] 张妍[2] 华婷[1] 张秋丽[1] 

机构地区：[1]内蒙古民族大学附属医院,内蒙古通辽028000 [2]吉林大学第二医院,吉林长春130041

出　　处：《中国实用眼科杂志》2016年第1期87-91,共5页Chinese Journal of Practical Ophthalmology

基　　金：内蒙古自治区自然科学基金（2011MS1166）

摘　　要：目的探讨EIA激活基因阻遏子（CREG）过表达修饰的骨髓间充质干细胞（BMSCs）能否有效对抗炎症因子过氧化氢（H2O2）诱导的BMSCs细胞凋亡，为其在视网膜变性类疾病的治疗提供理论基础。方法实验研究。对2012年1月至2014年12月在内蒙古民族大学附属医院眼科实验研究，以原代培养的大鼠BMSCs为实验细胞模型，以携带有CREG基因的腺病毒载体感染BMSCs，制备CREG基因修饰的BMSCs，Western blot及荧光显微镜鉴定CREG基因在BMSCs中的表达效率。TUNEL法检测添加500μmol／L过氧化氢（H2O2）刺激24h后，检测controlBMSCs组、normBMSCs组、GF—PBMSCs组、CREGBMSCs组的BMSCs凋亡率。Western blot方法检测caspase-3及BCL-2表达量。结果（1）瞬时转染Ad—GFP的BMSCs中GFP（绿色荧光蛋白）表达量达到90％，所构建的载体成功将目的基因CREG转染BMSCs。（2）通过TUNEL法和Western blot检测研究发现，Ad—CREG瞬时转染的BMSCs可以有效抑制500μmol／L H2O2刺激24h后诱导的凋亡，差异具有统计学意义（P〈0．05）。结论体外诱导BMSCs过表达CREG能有效抑制炎性因子H2O2诱导的BMSCs凋亡，对提高移植后BMSC存活率及今后应用于治疗视网膜变性类疾病可能发挥一定作用。Objective To ,determine whether rat bone marrow BMSCs transfected with CREG were able to effectively resist apoptosis induced by inflammatory mediators-hydrogen peroxide, which provide a theoretical basis in the treatment of relevant retinal degeneration diseases. Methods Our experimental study was based on the primary cultured rat bone marrow mesenchymal stem cells and made BMSCs modified with CREG in Adenovirus vector. Western blot and fluorescence microscope detect expression of efficiency of CREG in BMSCs. TUNLE investigate apoptosis rate of three groups （ normBMSCs, GFPBMSC, CREGBMSCs ） after hydrogen peroxide （H2O2） stimulating 24 hours. Apoptosis mark caspase-3 and BCL-2 was determined by western blot. Results The expression of GFP in BMSCs was over 90%. BMSCs were transfected with CREG successfully. The result of TUNLE and Western blot showed that BMSCs transfected with CREG could significantly inhibit apoptosis induced by 500μmol/L hydrogen peroxide （P 〈0.05）. Conclusions The overexpression of CREG can significantly inhibit apoptosis induced by hydrogen peroxide in vitro. So it may improve the survival rate of transplanted BMSCs and play an important role in the treatment of relevant retinal degeneration diseases.

关 键 词：骨髓间充质干细胞 E1A激活基因阻遏子 过氧化氢H2O2 细胞凋亡 

分 类 号：R543.5[医药卫生—心血管疾病]