黄芪多糖对脑缺血再灌注大鼠脑皮质中HSP70、PKB和P53蛋白表达的影响  

Effects of Astragalus polysaccharide on expression of HSP70,PKB and P53 in rat cerebral cortex with cerebral ischemia and reperfusion

作　　者：颜玲[1] 黄德彬[1] 刘锦红[1] 周庆华[2] 

机构地区：[1]湖北民族学院医学院,湖北恩施445000 [2]荆楚理工学院医学院,湖北荆州434000

出　　处：《中国病理生理杂志》2012年第9期1610-1617,共8页Chinese Journal of Pathophysiology

基　　金：湖北省自然科学基金资助项目(No.2008ABA197)

摘　　要：目的:探究黄芪多糖(AP)改善脑缺血再灌注大鼠神经功能和阻止脑皮质神经元凋亡的分子机制。方法:将120只雄性Wistar大鼠随机分成假手术组(SOG)、模型组(MG-1 d、3 d、7 d)、低剂量AP治疗组(L-APTG-1 d、3 d、7 d)和高剂量AP治疗组(H-APTG-1 d、3 d、7 d)。MG和APTG阻断右侧大脑中动脉形成缺血性脑损伤后,L-APTG和H-APTG分别腹腔注射AP 5 mg.kg-1和15 mg.kg-1。于1 d、3 d和7 d分别脑血流再灌注,神经功能缺损评分后处死取材,电镜观察神经元结构变化,流式细胞术分析神经元凋亡,免疫组化和West-ern blotting法检测脑皮质神经元热休克蛋白70(HSP70)、蛋白激酶B和P53蛋白表达。结果:H-APTG神经功能缺损评分和脑皮质神经元凋亡数显著低于MG和L-APTG(P<0.05);电镜下神经元结构(核糖体内质网、核仁、高尔基复合体、线粒体等)优于MG和L-APTG;在1 d、3 d和7 d,H-APTG脑皮质神经元HSP70和PKB蛋白表达显著高于L-APTG,后者又显著高于MG(P<0.05);H-APTG P53蛋白表达显著低于L-APTG,后者又显著低于MG(P<0.05)。结论:AP能改善脑缺血再灌注神经功能损伤和抑制神经元凋亡,其机制与促进脑皮质神经元HSP70和PKB蛋白表达,抑制P53蛋白表达有关。AIM：To investigate the effects of Astragalus injection on neuronal apoptosis and expression of c-Jun N-terminal kinase 3（JNK3） in the rat hippocampus after cerebral ischemia reperfusion. METHODS：The rat model of cerebral ischemia reperfusion was set up by a four-vessel occlusion method. The SD rats were randomly divided into 4 groups：sham operation group, cerebral ischemia reperfusion group（model group）, cerebral ischemia reperfusion＋Astragalus injection group（Astragalus injection group） and cerebral ischemia reperfusion＋vehicle group（vehicle group）. The rats in model group, Astragalus injection group and vehicle group after transient global cerebral ischemia（30 min） were then divided into 7 subgroups according to the reperfusion time of 0 h, 0.5 h, 2 h, 6 h, 24 h, 72 h and 120 h. The apoptosis of the neuron in the hippocampus was measured by the method of TUNEL staining. The expression of JNK3 at mRNA and protein levels was determined by real-time PCR and Western blotting,respectively. RESULTS：Compared with sham operation group, the number of apoptotic neurons increased in model group（P〈0.05）. Compared with model group, the number of apoptotic neurons decreased obviously in Astragalus injection group（P〈0.05）. Compared with sham operation group, the expression of JNK3 at mRNA and protein levels in the hippocampus increased obviously in model group at all time points except 120 h（P〈0.05）. Compared with model group, the expression of JNK3 at mRNA and protein levels in the hippocampus decreased obviously in Astragalus injection group at all time points except 120 h（P〈0.05）. CONCLUSION：Astragalus injection decreases neuronal apoptosis in rat hippocampus after cerebral ischemia reperfusion by inhibiting the expression of JNK3 at mRNA and protein levels.

关 键 词：脑缺血再灌注 黄芪多糖 大脑皮质 热休克蛋白70 蛋白激酶β 抑癌蛋白P53 

分 类 号：R363[医药卫生—病理学]