微创血肿抽吸术后大鼠脑出血周围组织细胞凋亡的研究

目的探讨脑出血周围组织神经细胞凋亡相关基因热休克蛋白70(HSP70)和半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)的变化规律及微创血肿抽吸干预治疗对神经细胞凋亡的影响。方法将SD大鼠75只分为假手术组、脑出血组、血肿抽吸组,各组又分为12 h、1 d、2 d、3 d、7 d 5个时间点。假手术组注入尾状核2μL生理盐水,脑出血组注入大鼠尾状核2μLⅣ胶原酶。血肿抽吸组是在脑出血模型制作6 h后进行血肿抽吸。各组分别于各时间点处死大鼠取脑组织,采用免疫组织化学染色测定各组大鼠脑出血周围HSP70、Caspase-3蛋白的表达并进行统计学处理。结果 HSP70、Caspase-3含量脑出血组较血肿抽吸组及假手术组均明显增高(P<0.05),血肿抽吸组比假手术组明显增高(P<0.05)。HSP70表达在12 h出现,3 d达高峰,随后表达逐渐减少。Caspase-3蛋白的表达在12 h出现,在3 d时达高峰,第7天表达明显减少。结论 Caspase-3蛋白高表达说明在脑出血第3天神经细胞凋亡和损害达到高峰,随后逐渐降低,HSP70高表达对出血脑组织具有明显的保护效应,且对Caspase-3蛋白的表达具有抑制作用,而且在早期对脑出血进行微创血肿抽吸干预治疗可以减少脑出血后神经细胞的凋亡。

Experimental research of apoptosis in rat tissue cells surrounding intracerebral hemorrhage after minimal hematoma aspiration

Objective To study the changes of apoptosis-related genes including HSP70 and caspase-3 in rat brains after experimental intracerebral hemorrhage and the effects of the minimally hematoma aspiration on neuronal apoptosis.Methods 75 SD rats were randomly divided into 3 groups the sham operated group,the intracerebral hemorrhage group and the hematoma aspiration group.Then each group was divided by time(12 h,1 d,2 d,3 d,and 7 d).2 μL normal saline was injected into the caudate nucleus in the sham operated group,2 μL Ⅳ collagenase was injected to the caudate nucleus in the intracerebral hemorrhage group.The intracerebral hemorrhage models were treated by the hematoma aspiration method six hours after they were produced.In this way,the hematoma aspiration group was made.The rats were killed at each time point and then their brain tissues were isolated.Expressions of HSP70 and caspase-3 in the cerebral tissues surrounding cerebral hemorrhage at different times were detected by immunohistochemical techniques and were statistically analyzed.Results HSP70 and caspase-3 expressions in the intracerebral hemorrhage group were significantly higher than those in the hematoma aspiration group and the sham operated group(P<0.05).Genes expressions in the hematoma aspiration group were significantly higher than those in the sham group(P<0.05).HSP-70appeared at 12 h,reached a maximum at 3 d,and then gradually decreased.Caspase-3 appeared at 12 h,reached a maximum at 3 d,and was obviously reduced at 7 d.Conclusions High expression of caspase-3 showed that nerve cells apoptosis and damage reached a peak at the third day after intracerebral hemorrhage,then gradually decreased.High expression of HSP70 has an obviously protective effect on intracerebral hemorrhage tissues,and reduces expression of caspase-3.The intervention of minimal hematoma aspiration in the early stage of cerebral hemorrhage can reduce the apoptosis of nerve cells.