摘要
目的观察大鼠大脑皮层神经元缺氧后细胞凋亡动态变化。方法制备大鼠大脑皮层神经元体外原代培养模型,免疫细胞化学鉴定大鼠大脑皮层神经元,透射电镜下观察不同时间点各实验组神经元超微结构的变化,TUNEL法观察不同时间点各实验组神经元凋亡情况。结果正常对照组神经元透射电镜下形态及染色质正常、内质网、线粒体等结构正常,缺氧组神经元水肿,细胞器破坏或消失;TUNEL法检测神经元凋亡:缺氧后各组神经元凋亡明显增加,与相应正常对照组相比有显著差异(P<0.01),缺氧48 h神经元凋亡达高峰(IOD为0.187±0.007),与缺氧12、24 h及72 h相比有显著差异(P<0.01)。结论细胞凋亡在缺血、缺氧性脑损伤中呈动态变化,是神经元死亡的重要形式。恰当时间窗内对神经元凋亡进行干预将是缺血、缺氧性脑病的有效治疗措施。
Objective To observe the dynamic changes of apoptosis in rat cerebral cortex neurons af- ter hypoxia. Methods Rat cerebral cortex neurons were primarily cultured from SD rats born within 24 h and then identified by immunocytochemical assay. Then the identified cells were cultured in the medium containing 100μmol/L CoC12 to simulate hypoxic condition. The cells cultivated in normal condition served as normal con- （ normoxia group）. Ultrastructural changes of the neurons were observed by transmission electron microsco- py （TEM）. Neuronal apoptosis were observed by TUNEL assay. Results TEM displayed that the morphology of neurons was normal, so was the structure of chromatin, endoplasmic reticulum and mitochondria in normoxia group, while, cellular edema, organelle damage or disappearance were seen in the hypoxia group. TUNEL showed that obvious apoptosis were found in hypoxic cells, with significant difference with normoxia group （P 〈 0. 01 ）. The apoptosis reached its peak in 48 h after hypoxia （0. 187 ±0. 007）, significantly higher than those in 12, 24 and 72 h （P 〈0. 01 ）. Conclusion Apoptosis is a dynamic process in hypoxic-ischemic brain inju- ry, and an important pattern of neuronal death. Intervention for neuronal apoptosis should be performed in an appropriate time window to effectively treat hypoxic-ischemic encephalopathy.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2012年第24期2489-2492,共4页
Journal of Third Military Medical University
基金
贵州省科学技术基金[(2007)2096号]
贵阳市科学技术计划项目[(2006)第26号]~~
关键词
神经元
缺氧
凋亡
电镜
neurons
hypoxia
apoptosis
electron microscope
