卡配因I细胞周期依赖性蛋白激酶5通路在血管性痴呆小鼠海马中.docVIP

卡配因I细胞周期依赖性蛋白激酶5通路在血管性痴呆小鼠海马中 　 作者：王天俊 吕佩源 王贺波 郭宗成 尹昱 张和振 刘昌林 梁翠萍 【摘要】 目的 观察脑缺血再灌注导致的血管性痴呆小鼠不同时间点海马组织卡配因I(Calpain I)细胞周期依赖性蛋白激酶5(Cdk5)通路表达的变化。方法 雄性昆明小鼠160只，随机分为假手术组(n=65)和模型组(n=95)。采用双侧颈总动脉反复缺血再灌注的方法制备血管性痴呆(VD)模型。术后第4周和第6周应用水迷宫实验进行行为学测试，免疫组化观察海马CA1区Calpain I的表达，RTPCR观察Cdk5的mRNA表达，Westernblot 检测Cdk5的蛋白表达。结果 术后第4周和6周时模型组小鼠的学习、记忆成绩均明显劣于假手术组小鼠(Plt;0.05)。术后第4周模型组海马CA1区神经元Calpain I表达的平均光密度值(0.090 0±0.010 0)较假手术组(0.033 2±0.004 3)显著增高(Plt;0.05)，Cdk5 mRNA表达(0.91±0.07)较假手术组(0.35±0.02)显著增高(Plt;0.05);术后第6周模型组海马Calpain I表达的平均光密度值(0.104 0±0.008 4)较假手术组(0.032 7±0.004 5)显著增高(Plt;0.05)，Cdk5 mRNA表达较假手术组显著增高(0.80±0.07 vs 0.53±0.05，Plt;0.05)。结论 Calpain ICdk5通路参与了脑缺血再灌注后小鼠VD的发生。 【关键词】 脑缺血再灌注;痴呆;卡配因;细胞周期依赖性蛋白激酶5 　　【Abstract】 Objective To observe the changes of the expression of Calpain I cyclindependent protein kinase 5 (Cdk5) pathway at different time points following cerebral ischemia and reperfusion. Methods 160 male Kunming mice were randomly divided into sham and model groups. Vascular dementia (VD) mice model was made by three times of transient ischemia and reperfusion of bilateral common carotid arteries. The behavioral tests were examined respectively at 4, 6 w post surgery. The expression of Calpain I in hippocampal CA1 was measured by immunohistochemistry staining. Cdk5 mRNA was examined by RTPCR. Western blot was used to evaluate Cdk5 protein expression. Results Compared with those in sham group， the abilities of learning and memory of animals in model group were damaged significantly(Plt;0.05) at 4, 6 w post surgery. At 4 w，the OD value of Calpain I in model group was higher than that of sham group(0.090 0±0.010 0 vs 0.033 2±0.004 3), and there was significant differene between the two groups(Plt;0.05). The expression of Cdk5 mRNA in model group was higher than that of sham group(0.91±0.07 vs 0.35±0.02 ，Plt;0.05). At 6 w，the OD value of Calpain I in model group was higher than that of sham group(0.104 0±0.008 4 vs 0.032 7±0.004 5，Plt;0.05). The expression of Cdk