血管性痴呆早期海马神经元的线粒体-PUMA损伤机制  

THE MECHANISM OF THE MITOCHONDRIA DAMAGE IN THE EARLY PERIOD OF VASCULAR DEMENTIA

作　　者：陈天智 王智惠 朱莹 张文丽 魏蕊 徐玉翔 张艳淑 王瑞敏 

机构地区：[1]华北理工大学医学实验研究中心神经生物学研究所,河北省唐山市063210 [2]华北理工大学预防医学院

出　　处：《中国煤炭工业医学杂志》2017年第5期566-570,共5页Chinese Journal of Coal Industry Medicine

基　　金：国家自然科学基金(编号:31171354;81671223);留学人员科技活动项目择优资助项目(编号:C2015005002);华北理工大学博士启动基金(编号:25638299);大学生创新项目(编号:X2015140)

摘　　要：目的阐明血管性痴呆早期海马神经元损伤的线粒体机制。方法雄性SD大鼠随机分为假手术组(Sham)和实验组(BCCAO后7、14、21d)。通过间隔1周永久性结扎双侧颈总动脉(BCCAO)制备血管性痴呆模型;Sham组大鼠暴露双侧颈总动脉,但不予以结扎。采用Western Blot技术和免疫荧光染色检测海马CA1区神经元线粒体凋亡途径的相关蛋白Bax、Bcl2、凋亡相关蛋白PUMA的蛋白表达的变化,以及线粒体功能蛋白Cyt C亚细胞分布的变化。结果 (1)与Sham组相比,BCCAO各组海马CA1区Bcl2蛋白表达明显降低,而Bax蛋白表达显著升高;(2)激光扫描共聚焦显微镜结果显示BCCAO各组海马CA1区神经元中Cyt C与线粒体标志性蛋白TOM20的共表达较Sham组明显减少;(3)PUMA蛋白于BCCAO后14、21d的表达较sham组明显升高,BCCAO后7d的PUMA表达略有增加,但无显著性差异。结论血管性痴呆早期神经元线粒体产生的损伤可能与PUMA介导的凋亡通路相关。Objective To illuminate the molecular mechanism of the mitochondria damage in the early period of vascular dementia （VD）. Methods Adult male Sprauge - Dawley rats were divided into two groups： sham control and VD early stage groups （7d, 14d and 21d after BCCAO）. VD animal model was performed by permanent bilateral common carotid artery occlusion （BCCAO） at one week interval. For sham control animals, the bilateral common carotid arteries were exposed but not to be legated. Bcl2, Bax, PUMA and mitochondrial functional protein Cyt C were detected using Western blot and immunofluorescence staining in hippocampal CA1 region. Results Compared with the sham group, in BCCAO groups （BCCAO 14d, 21d） protein expression of anti-apoptotic Bcl2 was obviously decreased that paralleled the increase of pro - apoptotic Bax protein. LSMC observation showed that co - localization of Cyt C with TOM20, a mitochondrial marker significantly reduced in BCCAO groups compared to sham animals. In BCCAO 14d and 21d animals, PUMA protein robustly increased and BCCAO 7d led to slightly enhance but no statistic difference compared to sham control. Conclusion Neuronal mitochondria damage in early stage of vascular dementia may be associated with the apoptosis pathway that was mediated by PUMA.

关 键 词：血管性痴呆 线粒体 凋亡 受p53基因上调表达的凋亡调控基因 

分 类 号：R543.4[医药卫生—心血管疾病]