运动训练对出血性脑损伤GAP-43基因及蛋白表达的影响

The effects of exercises on expression of GAP-43 and GAP-43mRNA after ICH in rats

目的:观察运动训练对脑出血大鼠生长相关蛋白(GAP-43)基因及其蛋白表达的影响。方法:实验动物用健康雄性SD大鼠160只。先将96只大鼠随机分为3组,实验组(出血后运动,n=32)、对照组(出血后不运动,n=32)、假手术组(无出血不运动,n=32)。各组又分为术后第7天,第14天,第21天,第28天共4个时相点,每个时相点4只用于免疫组化检测,4只用于RT-PCR检测。实验组大鼠于术后第72小时开始跑笼训练,其余大鼠在标准笼内自由活动。再将另外64只随机分为脑出血后第6小时,第12小时,第24小时,第48小时,第72小时,第7天,假手术组和正常组,每组4只分别用于免疫组化和RT-PCR检测。结果:①免疫组化结果:GAP-43阳性细胞表达为细胞胞浆着棕黄色,阳性细胞主要分布于血肿周围和大脑皮质的神经元,实验组于脑出血后第12小时开始上调,持续到第7天,与正常组和假手术组相比差异有显著性(P<0.05)。实验组于运动后第14天出现上调,第28天达高峰,与对照组相比差异有显著性(P<0.05),实验组和对照组与假手术组相比差异有显著性(P<0.01)。②RT-PCR结果:脑出血后第12小时GAP-43 mRNA表达一度升高,与对照组比差异有显著性(P<0.05),之后维持较高水平,直到第14—28天。实验组表达于第14—28天与假手术组相比差异有显著性(P<0.05),与对照组比(P>0.05)。结论:GAP-43参与了脑出血后神经可塑性的发生,运动训练可促进GAP-43基因及蛋白表达,从而改善神经功能。

Objective： To investigate the effects of exercises on expression of GAP-43（growth associated protein-43） and GAP-43mRNA of neuronal cells after intracerebral hemorrhage（ICH） in rats. Method： Ninety-six male SD rats （body weight, 270-300g） were divided into three groups, trial gronp（ICH and exercises group, n=32）, control group （ICH group, n=32） and sham operated group （no ICH without exercises, n=32）. The rats brains were removed at the 7^th d, 14^th d, 21^st d, 28^th d after operation. The other 64 rats were divided into 8 groups （the 6^th h, 12^th h,24^th h,48^th h,72^nd h,7^th d after ICH, sham operated and normal）. The activations of GAP-43 and GAP-43mRNA were measured by immunohistochemistry and RT-PCR. The rats in trial group began cage-running exercises at the 72^nd h after operation. The others lived in standard cages. Result：①The result of immunohistoehemistry： GAP-43-positive cells distributed around the hematoma and neuron of cortex. The number of GAP-43 -positive ceils was very little in sham-operation group. In trial group up-regulation of GAP-43 displayed at the 12^th h＇after ICH, until the 7^th d after ICH, and there was significant difference compared with sham operated group and normal group （P〈0.05）. There was up-regulation of expression of GAP-43 in trial group at the 14^th d-28^th d after operation, and there was significant difference in trial group and control group compared with sham operated group （P〈0.01）, but in trial group the expression of GAP-43 was on a higher level than that in control group （P〈0.05）. ②The result of RT-PCR： There was up-regulation of GAP-43mRNA at the 12^th＇ h after ICH in trial group, compared with control group （P〈0.05）. In trial group and control group there was higher expression at the 14^th-28^th d than that in shamoperated group（P〈0.05）, but in trial group there was no significant difference compared with control group （P〉0.05）. Conclusion：The results suggested that GAP-43 participated in neuronal plasticity after ICH, and exercises training could clearly up-regulate the expression of GAP-43 and GAP-43-mRNA and improve neurological function.