汉防己甲素对大鼠急性脊髓损伤后神经细胞凋亡和相关基因表达的影响  

Neuronal apoptosis and related gene expression in response to Tetrandrine after acute spinal cord injury in rats

作　　者：王远政[1] 田晓滨[1] 李波[1] 孙立[1] 韩伟[1] 汪雷[1] 张一[1] 

机构地区：[1]贵州省人民医院骨科,贵阳550002

出　　处：《中华创伤杂志》2012年第5期460-465,共6页Chinese Journal of Trauma

基　　金：贵州省省长基金资助项目（黔省专合字[2005】27号）

摘　　要：目的动态观察汉防己甲素（tetrandrine，Tet）对大鼠急性脊髓损伤（acutespinalcordinjury，ASCI）后神经细胞凋亡和相关基因表达的影响，为该病治疗研究提供实验依据。方法将95只成年SD大鼠随机分为空白对照组、急性脊髓损伤组（ASCI组）、汉防己甲素（Tet）组和甲基强的松龙组（Mp）组。空白对照组5只大鼠，其他每组30只大鼠。除空白对照组外，所有动物均用改良Allen重物打击法制作脊髓损伤模型。于术后8h、1，3，7，14和21d每组处死5只大鼠，观察损伤段脊髓的形态结构、细胞凋亡调控基因（Bcl—xl和Bcl—xs）的表达，运用流式细胞仪对标本进行凋亡细胞半定量分析。结果（1）组织学改变：除空白组无明显改变外，ASCI组组织水肿，神经元变性，部分坏死，可见大量炎性细胞浸润；Tet组损伤程度减轻；Mp组更轻，无明显坏死及炎性细胞浸润。（2）凋亡基因表达检测：除空白组外，余各组在相同时相点，Bcl—xs表达量ASCI组较多，Tet组明显减少，Mp组更少；而Bcl—xl表达量则反之，且各组在伤后7d达高峰。（3）凋亡细胞半定量分析：除空白对照组外，各组凋亡细胞数均在伤后7d达峰值，之后逐渐减少；同一时相点各组间凋亡细胞比较，Tet组明显少于ASCI组，而多于Mp组。结论ASCI后Tet对凋亡调控基因的表达有明显的积极干预作用，可显著减少神经细胞凋亡；应用Tet治疗ASCI应越早越好，7d内有效；与甲基强的松龙相比，Tet对ASCI的疗效较弱。Objective To analyze the effect of Tetrandrine （Tet） on neuronal apoptosis and related gene expression after acute spinal cord injury （ASCI） in SD rats so as to provide experimental basis for treatment of ASCI. Methods A total of 95 adult SD rats were randomly divided into four groups, ic, control group （n = 5 ） , ASCI group （n = 30） , Tet treatment group （Tet group, n = 30） and methylprednisolone treatment group （ Mp group, n = 30）. The ASCI model was established with the aid of modified Allen method. The five rats from each group were sacrificed at postoperative 8 hours and 1, 3, 7, 14, 21 days respectively to observe the morphology of the injury spinal cord and expressions of cell apoptosis regulation genes （Bcl-xl, Bcl-xs）. The flow eytometry （FCM） was used to semiquantitatively analyze the apoptotie cells. Results Except for the control group, the other three groups had morphologi- cal changes. Tissue edema, neuronal degeneration with some necrosis and marked inflammatory cell infiltration were founded in the ASCI group. Tissue injury was alleviated in the Tet group and was relatively much milder in the Mp group, with no obvious necrosis or inflammatory cell infiltration. Except for the control group, the number of Bcl-xs positive cells at the same time point was the most in the ASCI group in comparison with the Tet and Mp groups. Meanwhile, the expression of Bcl-xl in all groups reached the peak at day 7 post-injury. The semiquantitative analysis of the apoptotic cells displayed that, except for the control group, the number of apoptotic neurons in other three groups reached the peak at day 7 post- injury and then declined gradually. At the same time point, the apoptotic neurons in the Tet group were less than that in the ASCI group, but more than that in the Mp group. Conclusions After ASCI, Tet plays an active role in intervening the expression of the apoptosis regulation gene and can evidently decrease neuronal apoptosis. Tet treatment can work within 7 days and should be

关 键 词：脊髓损伤 神经元 粉防已碱 细胞凋亡 基因表达调控 

分 类 号：R285.5[医药卫生—中药学]