携带端粒酶反转录酶基因siRNA慢病毒载体对星形胶质细胞凋亡的影响  

siRNA lentiviral vectors carrying telomerase reverse transcriptase gene hasten astrocytes apoptosis

作　　者：梁亮[1] 徐韬[1] 宋扬[2] 盛伟斌[1] 

机构地区：[1]新疆医科大学第一附属医院脊柱外科,新疆维吾尔自治区乌鲁木齐市830054 [2]解放军第四军医大学唐都医院骨科,陕西省西安市710038

出　　处：《中国组织工程研究》2015年第11期1707-1711,共5页Chinese Journal of Tissue Engineering Research

基　　金：新疆维吾尔自治区自然科学基金资助项目面上项目(2014211C026);新疆维吾尔自治区自然科学基金资助项目青年基金(2014211C072)~~

摘　　要：背景:端粒酶反转录酶对端粒酶的激活和活化有重要的作用,利用端粒酶反转录酶的慢病毒载体抑制星形胶质细胞表达对脊髓损伤修复的影响鲜见报道。目的:利用靶向大鼠脊髓源星形胶质细胞端粒酶反转录酶基因的慢病毒载体转染大鼠星形胶质细胞,并观察端粒酶反转录酶基因的慢病毒载体对星形胶质细胞凋亡的影响。方法:原代及传代培养大鼠星形胶质细胞。实验分为端粒酶反转录酶基因siRNA慢病毒载体转染组、单纯慢病毒转染组和空白组,转染后并测其转染率,且在转染后的不同时间段测量大鼠星形胶质细胞的凋亡情况。结果与结论:端粒酶反转录酶基因siRNA慢病毒载体转染组及单纯慢病毒转染组对星形胶质细胞的转染率达85%-90%。免疫荧光染色及流式细胞仪检测显示,端粒酶反转录酶基因siRNA慢病毒载体转染组在转染后24-48h细胞凋亡率达50%-60%。在单纯慢病毒转染组及空白组细胞凋亡率并无显著改变。结果说明,携带端粒酶反转录酶基因siRNA慢病毒载体可促进大鼠脊髓星形胶质细胞凋亡。BACKGROUND：Telomerase reverse transcriptase plays an important role in telomerase activation, and lentiviral vectors carrying human telomerase reverse transcriptase that can inhibit astrocyte expression are rarely reported to have effects on spinal cord injury. OBJECTIVE：To transfect rat astrocytes with telomerase reverse transcriptase gene lentiviral vectors and to observe the effect of telomerase reverse transcriptase gene lentiviral vectors on apoptosis of astrocytes. METHODS： Astrocytes from rats were subject to primary culture and subculture, and then transfected with siRNA lentiviral vector carrying telomerase reverse transcriptase gene （siRNA transfection group）, simple lentiviral vector （lentiviral vector group） and nothing （blank group）, respectively. Then, the transfection efficiency and apoptosis in different time periods after transfection were determined. RESULTS AND CONCLUSION：The transfection efficiency was up to 85%-90% after siRNA lentiviral vector and simple lentiviral vector transfection. The apoptosis rate of astrocytes in the siRNA transfection group was 50%-60% at 24-48 hours after transfection, but there was no significant change in the other two groups. These findings suggest that siRNA lentiviral vectors carrying telomerase reverse transcriptase gene can accelerate astrocytes apoptosis.

关 键 词：基因 细胞 组织构建 组织工程 端粒酶 端粒酶反转录酶 慢病毒载体 RNA干扰 星形胶质细胞 转染 凋亡 脊髓损伤 胶质瘢痕 新疆维吾尔自治区自然科学基金 

分 类 号：R318[医药卫生—生物医学工程]