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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 cortical neurons after hypoxia. Methods Primary cultured rat cortical neurons were cultured in vitro. The neurons in the cerebral cortex of rats were identified by immunocytochemistry. The ultrastructural changes of neurons in each experimental group were observed under transmission electron microscope at different time points. Experimental group of neuronal apoptosis. Results The neurons in normal control group were normal in morphology and chromatin, the structures of endoplasmic reticulum and mitochondria were normal, the edema of neurons in edema group and the organelles were destroyed or disappeared. The apoptosis of neurons was detected by TUNEL method: (P <0.01). The apoptosis of neurons reached peak at 48 h (IOD = 0.187 ± 0.007), which was significantly higher than that of hypoxia at 12 h, 24 h and 72 h Compared to significant differences (P <0.01). Conclusions Apoptosis changes dynamically in ischemic and hypoxic brain injury and is an important form of neuronal death. Proper time window intervention of neuronal apoptosis will be an effective treatment for ischemic and hypoxic encephalopathy.