低氧预处理对缺氧-复氧后体外培养大鼠海马神经元Fos、Jun表达和神经元凋亡的影响

目的　观察低氧预处理对体外培养大鼠海马神经元缺氧 复氧后Fos、Jun表达和神经元凋亡的变化。方法　取培养 12d的两组 (对照组和低氧预处理组 )神经元 ,同时置于缺氧环境 (0 90l LN2 、0 10l LCO2 )中培养 4h后取出 ,置含 0 10l LCO2 和空气的培养箱内复氧培养 2 4和 72h。于不同时间取出 ,观察神经元存活数 ,并分别用抗Fos和Jun抗血清进行免疫组织化学染色 ,观察Fos、Jun表达阳性和阴性神经元数目 ,计数Fos和Jun表达神经元所占百分率。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 复氧后大鼠海马培养神经元中Fos、Jun表达阳性神经元百分率和凋亡神经元百分率均较缺氧前显著增加。经低氧预处理的海马神经元缺氧 复氧后Fos、Jun表达神经元和凋亡神经元百分率均较对照组明显减少。神经元损伤程度减轻 ,神经元存活数明显高于对照组。　结论　低氧预处理可使培养中海马神经元对缺氧产生耐受 ,减少缺氧 复氧后神经元Fos和Jun表达 ,减少缺氧 复氧后神经元凋亡。

EFFECTS OF HYPOXIC-PRECONDITIONING ON Fos AND Jun EXPRESSION AND APOPTOSIS IN CULTURED RAT HIPPOCAMPAL NEURONS AFTER ANOXIA- REOXYGENATION in vitro

Objective To study the effects of hypoxic preconditioning on the Fos and Jun expression and apoptosis in cultured rat hippocampal neurons after anoxia|reoxygenation. Methods The hippocampal neurons cultured for 12d from control and hypoxic|preconditioning groups were exposed to anoxia environment (0^90?l/L N-2 0^10?l/L CO-2) for 4h and than reoxygenated for 24h and 72h.The expression of Fos and Jun were revealed immunocytochemically using the antiserum against Fos and Jun.The apoptosis was examied using the terminal deoxynucleotidyl transferese|mediated biotinylated deoxyuridine triphosphate nickel and labeling(TUNEL) method and flow cytometric analysis. Results The increased of percentage of Fos and Jun positive neurons and apoptosis neurons in cultured hippocampal neurons after anoxia|reoxygenation than those in befor anoxia.The precentage of Fos and Jun positive neurons and apoptosis neurons were less in the hypoxic|preconditioning group than that in control aftre anoxia|reoxygenation.Conclusion Hypoxic|preconditioning increases tolerance of hippocampal neurons to anoxia and decreases the precentage in Fos and Jun expressing neurons and apoptosis neurons in hippocampus aftre anoxia|reoxygenation in vitro. ;[