摘要
目的:观察大鼠脑缺血再灌注后海马内Nemo样激酶(Nemo-like kinase,NLK)蛋白表达的变化及细胞定位。方法:60只成年雄性Sprague-Dawley大鼠随机分为假手术组和脑缺血再灌注组,其中脑缺血再灌注组依术后不同的检测时间点分为:1、4、8 h和1、3、7 d 6个亚组。采用中脑动脉闭塞法(middle cerebral artery occlusion,MCAO)建立脑缺血再灌注模型,用Westernblot方法检测脑缺血再灌注后大鼠海马组织中NLK,活化的Caspase-3蛋白的变化趋势,用免疫组化和免疫荧光来检测NLK蛋白在脑组织中的细胞定位及可能发挥的生物学行为。结果:在脑缺血再灌注的海马组织中NLK蛋白随着脑缺血再灌注时间的延长而呈现先增高后降低,后再升高的趋势,其中缺血再灌注后8 h NLK蛋白表达达到高峰,与假手术组相比,差异有统计学意义(P﹤0.05)。活化的Caspase-3的表达随着观察时间的延长递增,3 d达到峰值,与假手术组比较,差异有统计学意义(P﹤0.05),随后下降。NLK定位于脑组织中的神经元细胞尤其是海马CA1区的锥体神经元。结论:脑缺血再灌注后NLK蛋白的表达显著上调,提示NLK参与了成年大鼠脑缺血的发展。
Objective:To investigate the expression and cellular localization of Nemo-like kinase (NLK) in hippocampus of rat following cerebral ischemia reperfusion (IR). Methods: Sixty adult male Sprague-Dawley rats were randomly assigned to two groups: sham-operated group and IR group. According to different observation time after surgery, IR rats were individed into six subgroups (1 h,4 h,8 h,1 d,3 d and 7 d groups). A middle cerebral artery occlusion (MCAO) model in adult rats was constructed. The expression changes of NLK and active Caspase-3 in hippocampus of rat following cerebral IR were detected by Western blot. The cellular location and possible biological behavior of NLK was investigated by immunohistochemistry and immunofluorescence staining. Results:Western blot result revealed that NLK expression gradually increased,reached a peak at 8 h after cerebral IR,then decreased,and finally increased. There were significant differences in NLK expression between sham-operated group and IR group at 8 h after reperfusion (P 〈 0.05). Moreover,Caspase-3 expression progressively increased and reached a peak at the 3rd day,then decreased. There were significant differences in caspase-3 expression between these two groups at the 3rd day after reperfusion (P 〈 0.05). Immunofluorescence staining showed that NLK expression was principally located in neurons,especially pyramidal neurons of CA1 area in the hippocampus. Conclusion:The expression of NLK remarkably increased after cerebral IR, suggesting that NLK was involved in the development of cerebral ischemia in adult rats.
出处
《南京医科大学学报(自然科学版)》
CAS
CSCD
北大核心
2012年第7期903-907,共5页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家自然科学基金(81070275)
关键词
脑缺血再灌注
Nemo样激酶
海马
cerebral ischemia reperfusion
Nemo-like kinase
hippocampus