黄芪注射液抑制脑缺血再灌注大鼠海马组织Apaf-1表达

Astragalus injection inhibits expression of Apaf-1 in rat hippocampus after cerebral ischemia and reperfusion

目的:观察黄芪注射液对脑缺血再灌注大鼠海马组织凋亡蛋白酶激活因子1(Apaf-1)蛋白及其mRNA表达的影响。方法:将健康雄性SD大鼠120只随机分为假手术组、脑缺血再灌注组、黄芪注射液干预组和溶剂对照组。采用四血管阻断法制备大鼠脑缺血再灌注模型,除假手术组外其余3组根据再灌注不同时点再分为0 h、0.5 h、2 h、6 h、24 h、72 h和120 h 7个亚组,于再灌注相应时点提取脑组织。采用免疫组织化学和Western blot-ting检测大鼠海马组织Apaf-1蛋白的表达,RT-PCR法检测Apaf-1 mRNA的表达。结果:除0 h和120 h外,脑缺血再灌注组各个时点Apaf-1蛋白及mRNA表达均较假手术组增加(P<0.05);与脑缺血再灌注组相比,黄芪注射液干预组各个时点Apaf-1蛋白及mRNA表达均显著减少(P<0.05),而溶剂对照组各时点与脑缺血再灌注组相比则均无显著变化(P>0.05)。结论:黄芪注射液能抑制大鼠海马组织Apaf-1蛋白及mRNA表达,从而抑制脑缺血再灌注大鼠海马神经元的凋亡。

AIM ： To investigate the effect of Astragal~ injection on the expression of apoptotic protease-activa- ting factor 1 （ Apaf-1 ） in the hippocampus of global cerebral ische-mia-reperfusion rats. METHODS： Male SD rats were randomly divided into 4 groups with 30 each： sham operation group, cerebral ischemia-reperfusion group, cerebral ischemia- repeffusion ＋ Astragalus injection group, and cerebral ischemia-reperfusion ＋ vehicle group. The global cerebral ischemia- reperfusion model of the rats was established by d-vessel occlusion. The rats in cerebral ischemia-reperfusion group, cere- bral ischemia-reperfusion ＋ Astragalus injection group and cerebral ischemia-reperfusion ＋ vehicle group were further divided into 7 subsets, according to the reperfusion time of 0 h, 0.5 h, 2 h, 6 h, 24 h, 72 h and 120 h. After reperfusion, the brains were removed at the corresponding time points. The protein expression of Apaf-1 in hippocampal neurons was detec- ted by immunohistochemistry and Western blotting. The mRNA expression of Apaf-1 was observed by RT-PCR. RE- SULTS ： Compared with sham operation group, the expression of Apaf-1 at mRNA and protein levels at all time points ex- cept 0 h and 120 h increased obviously in cerebral ischemia-reperfusion group （P 〈 0.05 ）. Compared with cerebral ische- mia-reperfusion group, the expression of Apaf-1 at mRNA and protein levels at all time points except 0 h and 120 h de- creased obviously in cerebral ischemia-reperfusion ＋ Astragalus injection group （P 〈 0.05 ）. However, those in cerebral is- chemia-reperfusion ＋ vehicle group had no obvious change （ P 〉 0.05）. CONCLUSION： Astragalus injection inhibits the expression of Apaf-1 at mRNA and protein levels in hippocampus of global cerebral ischemia-reperfusion rats, thus inhibi- ting the apoptosis of hippocampal neurons.