跑台训练对脊髓损伤后大鼠小脑浦肯野细胞的影响

关键词：跑台运动训练  小脑  浦肯野细胞  脊髓损伤  凋亡

Keywords:running exercise training  cerebellum  Purkinje cells  spinal cord injury  apoptosis

摘要：

      摘要 目的：探讨跑台运动训练对脊髓损伤（SCI）后大鼠小脑细胞脑浦肯野细胞的影响及其机制研究。 方法：选取108只雌性SD大鼠，随机分成3组，包括假手术组、SCI组、SCI运动组。SCI运动组大鼠于术后开始跑台运动训练，BBB评分评定大鼠脊髓损伤后后肢运动功能。分别在术后第3天、第7天、第14天取小脑组织，通过HE染色、尼氏染色检测大鼠小脑浦肯野细胞数量和形态变化；免疫组化检测小脑中浦肯野细胞caspase-9、mGluR1表达情况；免疫荧光检测小脑中浦肯野细胞caspase-3、mGluR1表达情况；Western Blot检测小脑组织凋亡相关蛋白Bax、Bcl-2、Cyt-C、caspase-9、caspase-3蛋白表达水平变化。 结果：与假手术组比较，SCI组、SCI+TT组BBB评分均显著下降（P<0.05）；小脑组织中浦肯野细胞数量减少，体积缩小、失去正常形态；小脑中凋亡相关蛋白Bax、Cyt-C、活化caspase-9、活化caspase-3蛋白表达水平显著升高（P<0.05），Bcl-2表达显著降低；浦肯野细胞caspase-9和caspase-3表达增加，mGluR1表达降低（P<0.05）。与SCI组相较，SCI+TT组评分BBB评分结果比较无显著性差异（P>0.05）；小脑组织中浦肯野细胞数量、正常形态占比增加；凋亡相关蛋白Bax、Cyt-C、活化caspase-9 、活化caspase-3蛋白表达水平下降（P<0.05），Bcl-2表达升高；浦肯野细胞caspase-9和caspase-3表达下降，mGluR1表达水平显著升高（P<0.05）。 结论：跑台运动训练可通过线粒体凋亡途径抑制脊髓损伤后大鼠小脑浦肯野细胞的凋亡。

Abstract:

      Abstract Objective：To investigate the effect and mechanism of treadmill training on Purkinje cells in the cerebellum of the rats with spinal cord injury （SCI）. Method：Total of 108 female SD rats were randomly divided into 3 groups: the sham-operated group，the SCI group， and the SCI + treadmill training(TT) group. The rats in the SCI exercise group started treadmill training after surgery， and BBB scores were used to assess the hindlimb motor function of rats with spinal cord injury. The cerebellum tissues were collected on the 3， 7 and 14 day after surgery respectively， and the number and morphological changes of Purkinje cells in the cerebellum were detected by HE staining and Nissler staining. The expression of caspase-9 and mGluR1 in Purkinje cells was detected by immunohistochemistry； the expression of caspase-3 and mGluR1 was detected by immunofluorescence； the expression of apoptosis-related proteins Bax， Bcl-2， Cyt-C， caspase-9 and caspase-3 in cerebellar tissue was detected by Western Blot. Result：Compared with the sham-operated group， the BBB scores in SCI and SCI+TT groups were significantly decreased（P<0.05）. The number of Purkinje cells in cerebellum was reduced， and the size was reduced and lost the normal morphology. The expression levels of apoptosis-related proteins Bax， Cyt-C， activated caspase-9， and activated caspase-3 proteins in cerebellum were significantly increased（P<0.05）， and Bcl-2 expression was significantly decreased； caspase-9 and caspase-3 expression was increased and mGluR1 expression was decreased in Purkinje cells（P<0.05）. Compared with the SCI group， there was no significant difference in the BBB scores of the SCI+TT group（P>0.05）； the number and normal morphology of Purkinje cells in cerebellum increased； the expression levels of apoptosis-related proteins Bax， Cyt-C， activated caspase-9 and activated caspase-3 decreased（P<0.05）， and the expression of Bcl-2 increased； the expression levels of caspase-9 and caspase-3 in Purkinje cells were decreased， and the expression level of mGluR1 was significantly increased（P<0.05）. Conclusion：Treadmill training can reduce the apoptosis of Purkinje cells in the cerebellum of the rats with spinal cord injury through the mitochondrial apoptosis pathway.