缺氧-复氧对大鼠海马培养神经元Bcl-2、Bax表达和神经元凋亡的影响

目的 观察缺氧-复氧对体外培养海马神经元Bcl-2和Bax表达和神经元凋亡的影响。方法 取培养12d的海马神经元，置于恒温(36℃)密闭容器内，连续充以无氧气体90％(体积分数)N2、10％(体积分数)CO2]，在缺氧条件下继续培养4h后，再于常氧培养箱内复氧培养24h和72h。于不同时间观察神经元存活数，并分别用抗Bcl-2和Bax抗血清进行免疫组织化学染色，观察缺氧-复氧后大鼠海马培养神经元Bcl-2和Bax表达。并用原位末端标记(TUNEL)法和流式细胞术分别检测缺氧-复氧对体外培养海马神经元凋亡的影响。结果 缺氧-复氧后24～72h,海马神经元对Bcl-2的表达逐渐减弱，对Bax的表达逐渐增强，对Bax／Bcl-2比值逐渐增大，凋亡神经元百分率逐渐增多。结论 缺氧-复氧后24～72h神经元凋亡的发生与神经元Bcl-2表达逐渐减弱，Bax表达逐渐增强，Bax／Bcl-2比值逐渐增大有关。

Effects of Anoxia-reoxygenation on the Bcl-2 and Bax Expressions and Apoptosis in Cultured Rat Hippocampal Neurons

Objective To investigate the effects of anoxia-reoxygenation on Bcl-2 and Bax expressions and apoptosis in cultured rat hippocampal neurons. Methods The hippocampal neurons cultured for 12 d were exposed to anoxia environment (90% N2 + 10% CO2) for 4 h and then reoxygenated for 24 h and 72 h. The neurons were immunocytochemically stained using the antiserum against Bcl-2, or Bax and were detected by using the terminal deoxynucleotidyl transferese -mediated biotinylated deoxyuridine triphosphate nickel end labeling (TUNED method and flow cytometric analysis. The mean optical density of Bcl-2. or Bax expression neurons and the precentage of the apoptosis neurons were observed. Results The decrease of Bcl-2 expression and the increase of Bax expression correlated the higher percentage of the apoptosis neurons in cultured hippocampal neurons after anoxia-reoxygenation for 24-72 h than those before anoxia-rexoygenation. Conclusions The occurrence of neurons apoptosis is related to the decreased of Bcl-2 expression and the increased of Bax expression in cultured hippocampal neurons after anoxia-reoxygenation for 24-72 h.