补阳还五汤和依达拉奉联用对小鼠急性脑缺血损伤后脑保护机制的研究  

Neuroprotective effects of Buyang Huanwu Decoction combined with edaravone in the brain of mice with acute cerebral ischemia injury

作　　者：钟芳芳[1] 吴承龙[1] 孙新芳[1] 王赵伟[1] 

机构地区：[1]浙江省绍兴市人民医院神经内科,312000

出　　处：《中国神经免疫学和神经病学杂志》2016年第4期267-271,共5页Chinese Journal of Neuroimmunology and Neurology

基　　金：浙江省绍兴市公益性技术应用研究计划项目(2013B70076)

摘　　要：目的探讨补阳还五汤和依达拉奉联用对急性脑缺血损伤后神经细胞凋亡及凋亡相关蛋白表达的影响,探讨其可能的脑保护机制。方法将60只小鼠随机分假手术组、模型组、补阳还五汤组、依达拉奉组以及补阳还五汤+依达拉奉组,每组12只。采用改良线栓法制作小鼠大脑中动脉缺血再灌注模型,给予补阳还五汤及依达拉奉药物干预。分别于再灌注后1d和7d,采用TUNEL法观察小鼠脑皮质缺血区神经细胞凋亡率,采用免疫组化方法观察小鼠脑皮质缺血区B淋巴细胞瘤2基因(bcl-2)、bcl-2相关X蛋白(bax)和半胱氨酸蛋白酶3(caspase-3)表达的阳性细胞数。结果与假手术组比较,模型组小鼠脑皮质缺血区凋亡指数升高(P<0.01),且bcl-2、bax和caspase-3表达的阳性细胞亦均升高(P<0.01);经补阳还五汤和(或)依达拉奉干预后,各药物组小鼠脑组织的凋亡指数及bax和caspase-3阳性细胞均较模型组下降(P<0.01),而脑组织bcl-2阳性细胞均较模型组增加(P<0.01),且补阳还五汤+依达拉奉联合用药组较单一用药组改变明显(P<0.05)。结论补阳还五汤与依达拉奉联用能抑制脑缺血再灌注损伤后脑细胞中促凋亡蛋白bax、caspase-3的表达;促进具有神经元保护作用的bcl-2蛋白的表达,从而抑制神经细胞凋亡,协同发挥脑保护作用。Objective To study the effects of Buyang Huanwu Decoction （BYHWD）combined with edaravone （ED）on the apoptosis of neuron and expression of apoptosis related protein following cerebral ischemia-reperfusion （I/R）and further to discuss the mechanism of neuroprotection. Methods Sixty mice were randomly divided into the sham group,the I/R group,the BYHWD group,the ED group and the BYHWD＋ED group,12 mice in each group. Each group was further randomly divided into 1 d and 7 d groups. The middle cerebral artery occlusion/reperfusion model was established by the improved intraluminal filament technique in mice. BYHWD and ED interventions were applied. Immunohistochemistry method was used to analyze the expression of bcl-2,bax and caspase-3 positive cells,and TUNEL method was used to evaluate the neuronal apoptosis. Results Compared with the sham group,the apoptosis index of neuron increased in the I/R group （P〈0.01）,the bcl-2,bax and caspase-3 positive cells also increased （P 〈0.01）. After intervention in the drug groups,the apoptosis index declined （P 〈0.01）,the bcl-2 positive cells increased （P 〈0.01）,but the bax and caspase-3 positive cells decreased （P 〈0.01）. Above indexes of the BYHWD＋ED group were better than those of the single drug group （P 〈 0.05 ）. Conclusions The combination of BYHWD and ED can decrease the expression of bax,caspase-3 protein, and increase bcl-2 expression to inhibit the neuronal apoptosis and accelerate the recovery of neural function. It suggests that the two agents play synergistic roles in protecting the brain from cerebral ischemic reperfusion injury.

关 键 词：补阳还五汤 依达拉奉 脑缺血再灌注 细胞凋亡 caspase-3 bcl-2 BAX 

分 类 号：R743.3[医药卫生—神经病学与精神病学]