健脑方对SHRsp大鼠卒中后脑神经细胞凋亡的防治作用

Effect of Jiannao Decoction on Cerebral Nerve Cell Apoptosis of SHRsp Model Rat after Stroke

目的:观察急性脑卒中后大鼠脑神经细胞凋亡的变化及健脑方对凋亡的防治作用。方法:选择易卒中型自发性高血压(SHRsp)大鼠制作急性脑卒中动物模型,用健脑方干预治疗,观察造模前后大鼠神经系统症状、体征及大鼠海马组织病理结构的改变,应用原位末端脱氧核苷酸转移酶(TdT)标记(TUNEL)法、免疫组化法及固定化蛋白质印迹(Western blot)法观察大鼠造模及药物干预前后脑神经细胞凋亡的改变。结果:预防给药组大鼠卒中发生率明显低于模型组(P<0.01)。TUNEL 法显示模型组棕色颗粒在3天后达到高峰,应用健脑方干预后,棕色颗粒表达明显减少(P<0.05)。Western 结果显示,模型组 bcl-2、bax、caspase3在卒中后不同时相较正常组表达均增加。应用健脑方预防及治疗后均可使 bcl-2表达升高(P<0.05或 P<0.01),caspase3表达下降(P<0.05或 P<0.01);bax 在预防给药组表达下降(P<0.05或 P<0.01),与治疗给药组比较差异无显著性(P>0.05)。结沦:健脑方对脑卒中大鼠有明显的脑保护作用,其机制与抑制神经细胞凋亡及凋亡相关信号的传导有关。

Objective：To observe changes of cerebral nerve cell apoptosis of the rat after acute stroke and the preventive and treating effect of Jiannao Decoction on the apoptosis. Methods：SHRsp rats were used to establish the model of acute stroke, which were treated with Jiannao Decoction. Symptoms and signs of the nervous system and changes of pathological structures of the hipocampus were investigated before and after establishment of the model,and changes of cerebral nerve cell apoptosis before and after establishmeot of the model and medication were investigated with TUNEL,immunohistochamical and Western blot methods.Results： The incidence rate of stroke in the prevention group was significantly lower than that of the model group（P〈0. 01）.TUNEL showed that the brown granules in the model group reached the high peak at 48h, and after treatment of Jiannao Decoction, the brown granules decreased significantly （P〈0.05）Results of Western blot indicated that expressions of bcl-2,bax,caspase3 increased at different time after stroke as compared with those in the normal group ,and after prevention and treatment of Jiannao Decoction,expression of bcl-2 increased（P〈0. 05 or P〈0. 01） ,expression of caspase3 decreased（P〈0. 05 or P〈0. 01）,and expression of bax in the prevention group decreased （P〈0. 05 or P〈0. 01 ）but with no significant difference as compared with that of the treatment group （P〈0. 05）.Conclusion：Jiannao Decoction has significant protective action on brain in the rat with stroke,which is related with inhibitingthe effect of nerve cell apoptosis and conduction of apoptosis-related signals.