醒脑静注射液对颅脑外伤大鼠脑细胞凋亡及相关凋亡蛋白影响的研究

Study on the effects of Xingnaojing Injection on brain cell apoptosis as well as expressions of apoptosis-related proteins in rats with traumatic brain injury

目的探讨醒脑静注射液对颅脑外伤大鼠脑细胞凋亡及相关凋亡蛋白Fas、Fas-L、Caspase-3表达的影响。方法将108只SD健康成年大鼠(雌雄不拘)随机分为假手术组、损伤组、醒脑静组,每组36只。损伤组和醒脑静组采用Feeney's法造模,假手术组仅做颅骨开窗和清创缝合头皮。醒脑静组在脑打击后5 min内腹腔注射醒脑静注射液20 mL/kg,每天1次。3组大鼠分别在实验第1,3,5,7,14,21天取脑组织标本,免疫组化法检测Fas、Fas-L和Caspase-3表达情况,原位末端标记法(TUNEL)检测细胞凋亡情况。结果损伤组各时间点Fas、Fas-L、Caspase-3蛋白表达量和细胞凋亡率均明显高于假手术组(P均<0.05);醒脑静组各时间点Fas-L、Caspase-3蛋白表达量和第3—21天Fas蛋白表达量和细胞凋亡率均明显低于损伤组(P均<0.05)。结论 Fas、Fas-L和Caspase-3蛋白参与了颅脑外伤大鼠脑细胞损伤过程;醒脑静注射液干预可明显减少脑细胞凋亡,降低Fas、Fas-L和Caspase-3蛋白的表达,可能与醒脑静注射液影响信号转导的死亡受体通路有密切关系。

Objective It is to explore the effects of Xingnaojing Injection on the brain cell apoptosis and the expression levels of Fas,Fas-L,Caspase-3 in the rats with traumatic brain injury（ TBI）. Methods One hundred and eight SpragueDawley healthy adult rats were randomly divided into three groups： sham-operated group,TBI group,Xingnaojing group,each group had 36 rats. The models were established by Feeney’s method in TBI group and Xingnaojing group,skull opening and scalp debridement suture was only performed on in sham operated group. Xingnaojing group was treated with Xingnaojing Injection 20 m L/kg once per day by intraperitoneal injection in 5 min after injury. On 1 st,3 rd,5 th,7 th,14 th and21 stday,six rats in each group were used to assess brain cell apoptosis by TUNEL and the expression of fas/fas-L,Caspase-3 by immuno-histochemical staining. Results Compared with the sham-operated group,the expression of Fas,FasL and Caspase-3 protein at every time point and apoptotic rate of cells were significantly higher in TBI group（ P 〈 0. 05）;the expression of Fas-L and Caspase-3 protein at every time point and Fas protein from the 3 rd to 21 st day and apoptotic rate of cells were significantly lower in Xingnaojing group than that in TBI group（ P 〈 0. 05）. Conclusion Fas,Fas-L,Caspase-3 protein play roles in the development of cerebral injury in the rats with TBI,Xingnaojing injection could obviously decrease apoptosis of cerebral cells,inhibit the expression of Fas,Fas-L and Caspase-3 protein,the mechanism may be closely related with its effect on death receptor pathway of apoptosis.