摘要
目的 :观察携带Bcl 2基因的重组腺病毒 (Ad s Bcl 2 )对损伤运动神经元的保护作用。方法 :采用培养脊髓运动神经元的谷氨酸损伤模型 ,评价重组Bcl 2腺病毒对损伤运动神经元的影响。指标是Bcl 2原位杂交和Bcl 2免疫组化染色、TUNEL阳性神经元计数、运动神经元 [Ca2 +]i的检测以及台盼蓝拒染法检测培养神经元存活。结果 :①Ad s Bcl 2可转染原代培养的脊髓运动神经元并使其过表达Bcl 2。②过表达Bcl 2可延长培养神经元的生存时间。③过表达Bcl 2可显著减少谷氨酸诱导的原代培养脊髓运动神经元的凋亡 ,并显著降低谷氨酸诱导的神经元 [Ca2 +]i的增高。结论 :腺病毒中介Bcl 2基因过表达对神经毒性损伤的运动神经元具有保护作用。
Objective:To investigate the neuroprotective effect of Bcl 2, encoded by recombinant adenovirus (Ad/s Bcl 2), on injured motoneurons. Methods:The influence of the recombinant Bcl 2 adenovirus on injured motoneurons was estimated by using a glutamate induced lesion model. Bcl 2 in situ hybridization and immunohistochemical stain, TUNEL positive neuron counting, intracellular \[Ca 2+ \] i measurement and survival neurons testing by trypan blue dye exclusion technique were applied.Results: ① Administration of Ad/s Bcl 2 could transfer the primary cultured spinal cord motoneurons and make them overexpress Bcl 2. ② Overexpression of Bcl 2 could augment the survival of primary cultured rat spinal cord motoneurons in vitro . ③ Overexpression of Bcl 2 could strongly decrease the apoptosis and significantly alleviate intercellular \[Ca 2+ \] i increasing induced by glutamate. Conclusion:Overexpression of Bcl 2 mediated by adenovirus encoding Bcl 2 gene can protect motoneurons from neurotoxic injury.
