抑制钙蛋白酶对大鼠脊髓损伤后神经功能保护作用研究

Protective effect of Calpain inhibitor E-64-D on nerve function after rats spinal cord injury

目的:探讨钙蛋白酶抑制剂E-64-D对大鼠脊髓损伤后神经功能保护作用。方法:制备大鼠脊髓损伤模型,并将造模成功的大鼠随机分为脊髓损伤组(24只)和脊髓损伤干预组(24只)。另选取10只正常大鼠作为对照组,仅行假手术。脊髓损伤干预组在术后给予E-64-D静脉微量泵入。在术后1、7、14 d和28 d时(n=6)对大鼠脊髓Calpain-1 m RNA水平进行检测,并且对大鼠后肢功能进行评价。采用HE和Nissl染色法对大鼠伤处脊髓组织进行染色和评价。结果:对照组大鼠Calpain-1 m RNA水平在不同时间点之间无统计学差异,而脊髓损伤组和脊髓损伤干预组在不同时间点之间的差异均具有统计学意义,呈逐渐下降趋势(P<0.05)。在第28天时,脊髓损伤干预组和对照组Calpain-1 m RNA表达水平差异无统计学意义(P>0.05)。脊髓损伤组和脊髓损伤组干预组大鼠Tarlov评分和斜板实验结果在术后3个时间差异均具有统计学意义(P<0.05),并且脊髓损伤组干预组大鼠运动功能恢复情况显著优于脊髓损伤组大鼠。脊髓损伤组干预组大鼠神经元染色形态及数量均优于脊髓损伤组大鼠。结论:抑制钙蛋白酶能够抑制脊髓损伤后神经元的凋亡及坏死,促进脊髓功能恢复。

Objective：To investigate the protective effect of Calpain inhibitor E-64-D on nerve function after rat spinal cord injury.Methods：Rats spinal cord injury model was prepared by Allen＇s method. The successful models of rats with spinal cord injury were randomly divided into injury group（n=24）and intervention group（n=24）. Another 10 normal rats were selected as the control group,and only received sham surgery. Spinal cord injury intervention group were given E-64-D by intravenous injection in the post-operation. Rat spinal cord calpain-1 m RNA levels and hindlimb function were detected and evaluated on the 1,7,14 d and 28 d（n=6 at each time point）after operation. Rat tissues of spinal cord were evaluated by HE and Nissl staining. Results：There was no significant difference in Calpain-1 m RNA levels of control group at different time points,while differences existed between spinal cord injury group and spinal cord injury intervention group at different time points（P〈0.05）. On 28 d,Calpain-1 m RNA expression level of the spinal cord injury intervention group and control group had no significant difference（P〉0.05）. Tarlov score and inclined plane test results between pinal cord injury group and spinal cord injury intervention group had significant differences on the postoperative 7,14,28 d（P〈0.05）. Motor function of spinal cord injury intervention group was better than that of spinal cord injury group. Neurons form and quantity of spinal cord injury intervention group were better than those of spinal cord injury group. Conclusion：Inhibiting Calpain after spinal cord injury can prevent neuronal apoptosis and necrosis,and promote the recovery of spinal cord function.