c-jun反义基因重组体转染对缺氧复合烧伤血清刺激下大鼠心肌细胞的保护作用

Protective effects of c-jun anti-sense gene recombinant transfection on rat cardiomyocytes inflicted by hypoxia and burn serum

目的 观察c jun反义基因重组体转染对缺氧复合烧伤血清刺激下大鼠心肌细胞的保护作用。 方法 培养大鼠心肌细胞 ,分为 :(1)正常对照组 ;(2 )转染组 :构建c jun反义基因重组体并转染入心肌细胞 ,随后进行缺氧复合烧伤血清刺激 ;(3)非转染组 :仅进行缺氧复合烧伤血清刺激 ,不作其他处理。于刺激后 1、3、7h,采用逆转录聚合酶链式反应 (RT PCR)检测c junmRNA的表达变化 ;用Westernblot检测c jun蛋白、肌钙蛋白T(TnT)和 β 微管蛋白的表达变化 ;在光镜和电镜下观察心肌细胞形态结构改变。 结果 (1)与正常对照组比较 ,非转染组c junmRNA与蛋白表达显著 ,转染组较非转染组显著下降。 (2 )与正常对照组比较 ,非转染组TnT与 β 微管蛋白表达显著下降 ,转染组较非转染组明显回升。 (3)转染组心肌细胞骨架结构基本保持完好 ,偶见断裂与溶解。非转染组骨架网状结构紊乱、溶解与断裂 ,呈颗粒状。 结论 缺氧复合烧伤血清刺激可使大鼠心肌细胞c jun表达上调 ,进而引起心肌细胞损伤 ,c

Objective To investigate the protective effects of c-jun antisense gene recombinant transfection on rat cardiomyocytes inflicted by hypoxia and burn serum. Methods Cardiomyocytes from Wistar rat were isolated and cultured before being divided into normal control (C ), transfection (T) and non-transfection(NT) groups. C-jun antisense gene recombinant was constructed and transfected into cardiomyocytes, which were then treated by hypoxia and burn serum in T group, while those in NT group were simply treated by hypoxia and burn serum .The changes in the c-jun mRNA expression were determined by RT-PCR at 1,3 and 7 hours after the cardiomyocytes being stimulated, and the changes in the expressions of c-jun protein ,troponin-T(TnT) and β-tubulin were assayed by Western blot. The morphological changes in the cardiomyocytes were observed by LM and EM. Results 1)The expressions of c-jun mRNA and protein in the NT group were increased evidently when compared with those in C and T groups. 2) The expressions of TnT and β-tubulin in NT group were decreased evidently in contrast to those in C and T groups. In addition, there exhibited evident structural derangement, dissolution and fragmentation to granules of cardiomyocytes in NT group, while the myocardial cytoskeletal structure was well preserved with scarce fragmentation and dissolution in T group. Conclusion Increased expression of c-jun in rat cardiomyocytes resulting in myocardial injury could be induced by combined treatment of hypoxia and burn serum, while c-jun antisense gene recombinant transfection might protect rat cardiomyocytes from injury.