用发夹寡核苷酸探针检测新生大鼠脑缺氧缺血时细胞凋亡的变化(英文)  

Hairpin oligonucleotide probe as a marker of apoptotic change for cerebral hypoxia-ischemia in rat pups

作　　者：朱长连[1] 王小阳[1] 杨平[1] 程秀永[1] 

机构地区：[1]河南医科大学第三附属医院,郑州450052

出　　处：《河南医科大学学报》2001年第2期139-145,F003,共8页Journal of Henan Medical University

基　　金：国家自然科学基金资助项目! 3 0 0 70 2 71

摘　　要：目的 :观察用生物素标记的发夹寡核苷酸探针 (HPP)检测未成熟大鼠脑缺氧缺血后DNA双链断裂(DSB)的短暂空间改变 ,并与TdT介导的dUTP缺口末端标记 (TUNEL)的DNADSB比较。方法 :使 7d龄幼鼠缺氧缺血 ,用HPP位点标记显示DNADSB短暂空间形式 ,用微管相关蛋白 (MAP 2 )双标记评价HPP标记和脑损伤的关系 ,用双标记的TUNEL评价HPP标记的灵敏度。结果 :HPP标记局限在同侧半球MAP 2染色阴性区域 ;阳性细胞数与再灌注时间有关 ,在缺氧缺血后 2 4h ,阳性细胞数上升达到高峰 ,但在松果体细胞核变化更早 ,而海马CA3 区域细胞变化延迟。HPP和TUNEL双标记显示在再灌注早期HPP标记多于TUNEL标记。结论 :HPP标记显示出缺氧缺血后不同时间点凋亡细胞的典型变化 ,是检测凋亡改变的灵敏标记。Aim: To investigate temporal and spatial changes of DNA double-stranded break (DSB) detected by biotin-labeled hairpin oligonucleotide probe(HPP) and compare with TdT-mediated dUTP nick end labeling (TUNEL) for detectiog DNA DSB in the immature rat brain after hypoxia-ischemia. Methods: 7-day-old rat pups were subjected to hypoxia-ischemia. The spatiotemporal pattern of DNA DSB was characterized with HPP in situ labeling. The relation of HPP labeling and brain damage was evaluated by double labeling with microtubule associated protein 2 (MAP 2). The sensitivity of HPP labeling was evaluated by double labeling with TUNEL.Results: HPP labeling localized in the ipsilateral hemisphere within the area of loss MAP 2 staining. The number of positive cells with nucleus condensation and cytoplasm shrinkage were remarkable with prolongation of reperfusion. The number of positive cells increased and reached peak at 24h post-HI. But in the nucleus habenularis displayed a faster time course. Hippocampus CA 3, however, shows a more delayed time course. Double labeling of HPP and TUNEL showed more HPP labeling than TUNEL in the early reperfusion.Conclusion: HPP labeling shows typical apoptotic nucleus changes with different timepoints after HI. It is a sensitive marker for detecting apoptotic changes.

关 键 词：脑缺氧 脑缺血 细胞凋亡 双链断裂 大鼠 生物素标记 发夹寡核苷酸探针 

分 类 号：R743.31[医药卫生—神经病学与精神病学]