c-fos反义基因转染对缺氧复合烧伤血清处理大鼠心肌细胞保护作用的实验研究  

Effect of c-fos antisense gene transfection on protection of cardiomyocytes following hypoxia and burn serum treatment in rats

作　　者：胡安根[1] 黄跃生[1] 

机构地区：[1]第三军医大学附属西南医院全军烧伤研究所

出　　处：《第三军医大学学报》2003年第18期1599-1601,共3页Journal of Third Military Medical University

基　　金：国家重点基础研究发展规划资助项目 ("973"项目 ) (G19990 54 2 0 2 ) ;国家杰出青年科学基金资助项目 ( 30 12 50 4 0 ) ;全军"十五"指令性课题 ( 0 1L0 6 6 ) ;教育部高等学校骨干教师资助计划项目 ( 2 0 0 0 ) ;军队首批临床高新技术重大项目 ( 2 0 0

摘　　要：目的　探讨c fos反义基因转染对缺氧复合烧伤血清处理心肌细胞的保护作用。方法　烧伤血清采自 3 0 %TBSAⅢ度烫伤Wistar大鼠 ;采用含 1%O2 的混合气体作为缺氧模型 ;采用基因重组技术构建c fos反义基因重组体 ;由Li pofectaminekit介导心肌细胞转染 ;于缺氧复合烧伤血清处理 1,3 ,7h不同时相点采用RT PCR检测c fosmRNA的表达变化 ,采用Westernblot检测c fos蛋白 ,肌钙蛋白 T(TnT)和 β 微管蛋白 ( β Tubulin)的表达变化。 结果　加入烧伤血清并造成缺氧的非转染心肌细胞 (下文称非转染组 ) ,c fosmRNA和蛋白显著表达 ,3h达到最高峰 ,与正常对照组比较 ,TnT和 β Tu bulin表达显著下降 ,7h降至最低水平 ;转染c fos反义基因重组体 (下文称转染组 )后 ,c fosmRNA和蛋白表达显著下降 ,TnT和 β Tubulin表达却显著回升。 结论　缺氧复合烧伤血清处理产生的c fos会引起心肌细胞损伤 ,c fos反义基因重组体转染对缺氧复合烧伤血清处理心肌细胞具有保护作用。Objective To explore the effect of c fos antisense gene transfection on the protection of cardiomyocytes following hypoxia and burn serum treatment. Methods Burn serum was collected from Wistar rats with 30% total body surface area(TBSA) of Ⅲ degree burns. The mixture gas containing 1% O 2 was used as hypoxia model. The c fos antisense gene recombinant was constructed by genetic recombination technique. Cardiomyocytes from neonatal Wistar rats were cultured in vitro with hypoxia and burn serum treatment. c fos antisense gene recombinant was transfected into the cultured cardiomyocytes. Expression of c fos mRNA was determined by RT PCR. Expressions of c fos protein, troponin T and β Tubulin in cardiomyocytes were determined by Western blotting in the transfected and non transfected groups. Results RT PCR results showed that the expression of c fos mRNA increased significantly in the non transfected group. But after transfection of c fos antisense gene recombinant, the expression of c fos mRNA decreased significantly as compared with the non transfected cardiomyocytes. Western blotting results showed that the expression of c fos protein in the transfected group decreased remarkably as compared with the non transfected group, but the expressions of β Tubulin and troponin T increased significantly in the transfected group. Conclusion Burn serum and hypoxia can cause the injury of cardiomyocytes. c fos antisense gene recombinant transfection has the protective effect on cardiomyocytes exposed to burn serum and hypoxia.

关 键 词：β—微管蛋白 肌钙蛋白—T 反义基因 缺氧 烧伤血清 C-FOS 基因转染 PCDNA3.1(+) 

分 类 号：R322.11[医药卫生—人体解剖和组织胚胎学] R394.2[医药卫生—基础医学]