清脑益元汤对脑缺血损伤大鼠缺血侧皮质区神经细胞凋亡及凋亡相关蛋白Bcl-2、Bax表达的影响

Effect of QingnaoYiyuan decoction on neuronal apoptosis and expression of apoptosis-related proteins Bcl-2 and Bax in ischemic cortex of rats with cerebral ischemia injury

目的探讨清脑益元汤对脑缺血损伤大鼠缺血侧皮质区神经细胞凋亡及凋亡相关蛋白B淋巴细胞瘤(Bcl)-2、Bcl-2相关x蛋白(Bax)表达的影响及其可能的作用机制。方法将240只健康SD大鼠随机分为两大组,每大组再分为4个亚组:空白组、假手术组、模型组、清脑益元汤组。采用改良的Longa线栓法制备大鼠急性脑缺血损伤模型,按术后取材时间点分为5个亚组,即1、3、7、14、28 d 5个取材时间点。造模成功后取大鼠缺血侧脑组织,采用TUNEL法检测大鼠缺血侧皮质区神经细胞凋亡表达,采用Western印迹法和实时荧光定量PCR(Real-Time PCR)法检测大鼠缺血侧皮质区Bcl-2、Bax蛋白及mRNA表达。结果同一时间点,与假手术组相比,模型组TUNEL阳性凋亡细胞表达明显升高(P<0.01),Bcl-2蛋白及mRNA表达明显降低(P<0.01),Bax蛋白及mRNA表达明显升高(P<0.01);同一时间点,与模型组相比,清脑益元汤组阳性凋亡细胞表达降低(P<0.01);Bcl-2蛋白及mRNA表达升高(P<0.01),Bax蛋白及mRNA表达降低(P<0.01);结论清脑益元汤能有效抑制神经元凋亡,对脑缺血损伤具有保护作用,其作用机制可能与上调Bcl-2蛋白及mRNA表达,下调Bax蛋白及mRNA表达有关。

Objective To explore the effect of QingnaoYiyuan decoction on neuronal apoptosis and the expression of apoptosis-related proteins Bcl-2 and Bax in ischemic cortex of rats with cerebral ischemia injury and its possible mechanism.Methods 240 healthy SD rats were randomly divided into two groups,and each group was further divided into four subgroups:blank group,sham group,model group and QingnaoYiyuan decoction group.The rat model of acute cerebral ischemia injury was made by modified Longa suture method,which was divided into five subgroups according to the time points of postoperative sampling,named 1,3,7,14 and 28 days.After the model was successfully established,the ischemic brain tissue of rats was taken,the apoptosis expression of nerve cells in ischemic cortex was detected by TUNEL method,and the expressions of Bcl-2,Bax protein and mRNA in ischemic cortex were detected by Western blot and Real-time PCR.Results At the same time point,compared with the sham group,the expression of TUNEL positive apoptotic cells in model group increased significantly(P<0.01),the expressions of Bcl-2 protein and mRNA decreased significantly(P<0.01),and the expressions of Bax protein and mRNA increased significantly(P<0.01).At the same time point,compared with model group,the expression of positive apoptotic cells in QingnaoYiyuan decoction group decreased(P<0.01),the expressions of Bcl-2 protein and mRNA increased(P<0.01),while the expressions of Bax protein and mRNA decreased(P<0.01).Conclusions QingnaoYiyuan decoction could effectively inhibit neuronal apoptosis and protect against cerebral ischemia injury,and its mechanism might be related to up-regulating expressions of Bcl-2 protein and mRNA and down-regulating expressions of Bax protein and mRNA.