缺氧/复氧对培养的大鼠海马神经元Fos、Jun表达和神经元凋亡的影响

EFFECTS OF ANOXIA/REOXYGENATION ON FOS AND JUN EXPRESSION AND APOPTOSIS IN CULTURED RAT HIPPOCAMPAL NEURONS

目的 :观察缺氧 /复氧对体外培养的海马神经元Fos和Jun表达和神经元凋亡的影响。方法 :取培养 12d的海马神经元 ,置于 2 0 0 0cm3 的恒温 (36℃ )密闭容器内 ,连续充以无氧气体 (90 %N2 、10 %CO2 ) ,在缺氧条件下继续培养 2、4h后取出 ,置含 10 %CO2 和空气的培养箱内复氧培养 2 4h和 72h。于不同时间取出 ,观察神经元存活数 ,分别用抗Fos和抗Jun抗血清进行免疫组织化学染色 ,计数Fos和Jun表达阳性神经元百分率 ,并用原位末端标记 (TUNEL)法和流式细胞术分别观察和测定缺氧 /复氧对体外培养海马神经元凋亡的影响。结果 :缺氧 /复氧后Fos和Jun表达阳性神经元百分率和凋亡神经元百分率均显著增加。结论 :缺氧 /复氧后即早反应基因fos在神经元中的持续表达可引起神经元凋亡 。

Aim:To investigate the effects of anoxia/reoxygenation on Fos and Jun expression and apoptosis in cultured rat hippocampal neurons. Methods: The hippocampal neurons cultured for 12 d were exposed to anoxia environment (90% N 2+10% CO 2) for 4 h and then reoxygenated for 24 h and 72 h. The neurons were immunocytochemically stained using the antiserum against Fos and Jun, and the apoptosis were detected by using the terminal deoxynucleotidyl transferase-mediated biotinylated deoxyuridine triphosphate nickel end labeling(TUNEL)method and flow cytometric analysis. Results:The percentage of Fos and Jun positive neurons and apoptosis neurons in cultured hippocampal neurons after anoxia/reoxygenation increased than those in control. Conclusion:The occurrence of neurons apoptosis is related to the increase in Fos and Jun expression in cultured hippocampal neurons after anoxia/reoxygenation.