选择性钙蛋白酶抑制剂E-64-d对大鼠急性脊髓损伤的作用

发布时间：2018-09-14 11:21

【摘要】：[目的]利用急性脊髓损伤SD大鼠动物模型,探讨E-64-d在脊髓损伤修复过程中的作用机制。[方法]制作大鼠脊髓损伤模型,将大鼠随机分为5组:对照组;单纯损伤组;E-64-d治疗组;MP治疗组;MP与E-64-d联合治疗组。给与钙蛋白酶抑制剂E-64-d和/或甲基强的松龙(MP)腹腔注射。分别应用TUNEL法检测神经细胞的凋亡水平。免疫组化测定各时间点Calpain1与GFAP的表达变化;每天同一时间对大鼠进行后肢运动功能BBB评分及感觉功能评分。[结果]TUNEL检测显示,治疗组神经元细胞凋亡减少,Caspase-3的活性均减弱。损伤后脊髓组织中Calpain 1活性增加,应用E-64-d治疗后得到抑制。损伤后3 d Calpain 1和GFAP的表达增强,但治疗组显著低于损伤组。BBB运动功能评分显示,治疗组大鼠的运动功能得到增强,结果具有统计学意义。进一步实验结果显示,联合使用E-64-d和甲基强的松龙(MP)的效果要好于单独使用E-64-d或MP。[结论]E-64-d具有减轻急性大鼠脊髓损伤的作用,其与MP联合使用效果更加明显,Calpain抑制剂具有治疗脊髓损伤的潜力。
[Abstract]:[objective] to investigate the mechanism of E-64-d in the repair of spinal cord injury (sci) using acute spinal cord injury (SD) rat model. [methods] Spinal cord injury model was established in rats. The rats were randomly divided into five groups: control group, simple injury group, E-64-d treatment group and combined treatment group of MP and E-64-d. Calpain inhibitor E-64-d and / or methylprednisolone (MP) were given intraperitoneally. The level of neuronal apoptosis was detected by TUNEL method. The expression of Calpain1 and GFAP were measured by immunohistochemistry, and the BBB score and sensory function score of hindlimb motor function were evaluated at the same time every day. [results] TUNEL showed that apoptosis of neuronal cells decreased and the activity of Caspase-3 decreased in the treatment group. The activity of Calpain 1 in spinal cord tissue increased after injury and was inhibited after treatment with E-64-d. The expression of Calpain 1 and GFAP increased 3 days after injury, but the motor function score of treatment group was significantly lower than that of injury group. Further experimental results show that the combination of E-64-d and methylprednisolone (MP) is better than E-64-d or MP. alone. [conclusion] E-64-d can attenuate acute spinal cord injury in rats. The effect of E-64-d combined with MP is more obvious. Calpain inhibitor has the potential to treat spinal cord injury.
【作者单位】：上海第二军医大学第三附属医院;上海市光华中西医结合医院;
【基金】：长宁区科委重点项目资助(项目编号:CNKW2013Z01)
【分类号】：R651.2

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