川芎嗪对急性脊髓损伤大鼠神经细胞早期凋亡和Caspase-3mRNA表达的影响  

Effects of Tetramethylpyrazine on Expression of Early Apoptosis and Caspase-3 Gene in Rats With Spinal Cord Injury

作　　者：王健[1] 贾玉柱[2] 郑纯威[3] 徐卫星[4] 刘建[4] 

机构地区：[1]浙江中医药大学,浙江杭州310053 [2]浙江省立同德医院影像科,浙江杭州310012 [3]浙江省立同德医院实验动物中心,浙江杭州310012 [4]浙江省立同德医院骨科,浙江杭州310012

出　　处：《中华中医药学刊》2011年第12期2662-2668,共7页Chinese Archives of Traditional Chinese Medicine

基　　金：浙江省中医药管理局基金资助项目(2010ZB017)

摘　　要：目的:观察川芎嗪对大鼠急性脊髓损伤后神经细胞的早期凋亡及其对Caspase-3mRNA表达的影响,进一步认识Caspase-3在脊髓损伤后细胞凋亡中的作用机制。方法:采用改良Allen's重物打击法建立大鼠急性脊髓损伤模型,120只成年SD大鼠,随机分成4组:正常组、模型组、甲强龙及川芎嗪组,每组各30只,造模后模型组、甲强龙组和川芎嗪组分别按时注射同体积的生理盐水,甲强龙及川芎嗪注射液。损伤后各时间采用BBB法行神经功能评分,流式细胞术观察细胞凋亡情况,透射电镜观察细胞形态学变化,并应用RT-PCR技术检测Caspase-3mRNA表达的情况。结果:模型组伤后6h出现了神经细胞的早期凋亡,至伤后24h达到高峰,伤后48h比24h略有下降,但仍持续在较高的水平,差异无统计学意义(P>0.05);川芎嗪治疗组伤后各时间点细胞凋亡与甲强龙组伤后各时间点细胞凋亡比较差异无统计学意义(P>0.05),但川芎嗪组和甲强龙组与模型组比较差异有统计学意义(P<0.05)。伤后各时间点Caspase-3mRNA表达的趋势相近,都具有"量———时间"的依赖关系,川芎嗪治疗组与甲强龙组伤后各时间点Caspase-3mRNA比较差异无统计学意义(P>0.05),但川芎嗪组和甲强龙组分别与模型组比较差异有统计学意义(P<0.05)。结论:本研究发现大鼠SCI后神经细胞6h内的即出现早期凋亡,比电子显微镜下判断凋亡出现的时间要明显提早;SCI后,损伤区脊髓组织Caspase-3mRNA水平明显升高,与细胞凋亡的比例和细胞超微结构的变化具有明显的相关性;早期使用川芎嗪能减轻大鼠急性脊髓损伤后的神经细胞凋亡,对大鼠急性脊髓损伤有一定神经保护作用,主要机制可能和抑制细胞凋亡的"级联反应"有关。Objective： Creating acute spinal cord injury （SCI） model in rats with modified Alleng method, to observe morphology of nerve cells in early apoptosis and study caspase -3mRNA expression in rats with tetramethylpyrazine （TMP） after spinal cord injury, in order to further understand mechanism of action of Caspase -3 in apoptosis after SCI and to explain mechanism of TMP in inhibiting apoptosis at the gene level. Methods ： 120 adult SD rats were randomly divided into 4 groups： normal group, model control group, MP treatment group and the TMP treated group, 30 rats in each group. By modified Alleng method of making weight spinal cord injury model was established, after the SCI model was made, MP and TMP group were injected the same volume of saline on time. Neurological scores （BBB method） were recorded at different time points after injury. Apoptosis and morphological changes were observed by flow cytometry ease and by transmission electron microscopy. While Caspase - 3mRNA expression were detected by RT - PCR at different time points after injury. Results ： Apoptosis rates of nerve cells of model group appeared in early apoptosis in 6h, Apoptosis rates compared 48h with 24h , decreased slightly, but still sustained at a high level, the difference was not statistically significant （ P 〉 0.05 ）. The difference of apoptosis rates between TMP and MP treatment group at each time point was not statistically significant （ P 〉 0.05 ）, but compared with model group, the difference was statistically significant （ P 〈0.05）. Caspase -3mRNA expression was similar at different time points after injury, both have a ＂volume - time＂ dependent relationship. The difference of apoptosis rates and Caspase - 3mRNA both TMP group and MP group at different time points after injury was not statistically significant （P 〉 0.05）, but the difference compared the TMP group and the MP group with the model group were significant （P 〈 0.05）. Conclusion ： The study found that the early apoptos

关 键 词：脊髓损伤 大鼠 川芎嗪 细胞凋亡 CASPASE-3 基因 

分 类 号：R285.5[医药卫生—中药学]