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目的观察汉防己甲素(tetrandrine,Tet)对大鼠急性脊髓损伤后bcl-2和bax表达的影响,探讨其对脊髓损伤的作用机制。方法成年SD大鼠100只,分为4组:假手术组10只,损伤对照组、甲基强的松龙(methylprednisolone,MP)治疗组、Tet治疗组各30只。胸8、9椎板切除后,损伤对照组,MP、Tet治疗组用加速压迫型Allen’s打击法制成脊髓损伤模型,后2组动物于制模前,伤后24、48h尾静脉分别注射MP90mg/kg和1%Tet22.5mg/kg。各组大鼠于术后8h,1、3、7、14d行运动功能BBB评分,取损伤段脊髓行石蜡切片HE染色,观察脊髓组织的形态结构变化,免疫组织化学染色检测细胞凋亡因子bcl-2、bax表达。结果伤后7、14dMP、Tet治疗组大鼠运动功能评分显著高于损伤对照组(P<0.05),各时间点MP、Tet治疗组评分无统计学意义(P>0.05);MP、Tet治疗组脊髓组织损害较损伤对照组轻,术后8h至14d动态观察,3～7d损伤表现最为严重,达到损伤高峰期;假手术组中bax、bcl-2阳性细胞数较少,MP、Tet治疗组bax阳性细胞数少于损伤对照组(P<0.05),而bcl-2阳性细胞数多于损伤对照组(P<0.05)。结论 Tet可通过增加bcl-2表达和降低bax表达来抑制急性脊髓损伤后神经细胞的凋亡,有益于脊髓组织的保护,促进运动功能的恢复。
Objective To investigate the effect of tetrandrine on the expression of bcl-2 and bax after acute spinal cord injury in rats and its mechanism of action on spinal cord injury. Methods 100 adult SD rats were divided into 4 groups: sham operation group (n = 10), injury control group, methylprednisolone (MP) treatment group and Tet treatment group (n = 30). Thoracic 8, 9, lamina resection, injury control group, MP, Tet treatment group with accelerated compression type Allen’s hit the spinal cord injury model, the latter two groups of animals in the model 24 hours after injury, 48 h tail vein injection of MP90mg / kg and 1% Tet22.5 mg / kg. The BBB score of motor function was measured at 8h, 1,3,7,14d after operation in rats in each group. HE staining was performed on paraffin sections of injured spinal cord to observe the morphological changes of spinal cord. The expression of apoptosis-inducing factor bcl -2, bax expression. Results The scores of motor function in MP and Tet groups were significantly higher than those in injured control groups at 7 and 14 days after injury (P <0.05). There was no significant difference between MP and Tet groups at each time point (P> 0.05) Group spinal cord tissue damage than the damage control group, dynamic observation after 8h to 14d, 3 ~ 7d damage the most serious, reaching the peak of injury; sham-operated group less bax, bcl-2 positive cells, MP, Tet treatment The number of bax positive cells was less than that of injured control group (P <0.05), but the number of bcl-2 positive cells was more than that of injured control group (P <0.05). Conclusion Tet can inhibit the apoptosis of nerve cells after acute spinal cord injury by increasing the expression of bcl-2 and decreasing the expression of bax, which is beneficial to the protection of spinal cord tissue and the recovery of motor function.