持续低氧对海马神经元凋亡及蛋白表达影响

目的观察持续低氧致体外培养SD乳大鼠海马神经元细胞凋亡及c-jun、c-fos的表达情况。方法海马神经元原代细胞培养,取培养10 d的海马神经元,置于90%N₂、5%O₂、5%CO₂混合气体培养箱中培养2,6,12,24h后,Hoechst 33258染色观察神经元细胞凋亡情况,并用蛋白印迹法检测c-jun、c-fos蛋白表达水平。结果随缺氧时间延长,凋亡细胞所占比例由(23.79±3.43)%上升至(74.36±5.58)%;c-jun、c-fos蛋白在缺氧2 h时表达增高,6 h时达高峰,之后逐渐降低,24 h时仍略高于正常对照。结论持续低氧可引起体外培养海马神经元c-jun、c-fos蛋白表达升高,这可能与神经细胞凋亡以及保护机制有关。

Apoptosis and expressions of c-jun,c-fos in rat hippocampal cells in vitro with continuous hypoxia

ObjectiveTo observe the cell apoptosis and the expressions of c-jun,c-fos in neonatal SD rat hippocampal cells with continuous hypoxia in vitro.MethodsAfter ten-days primary culture,the hippocampal cells were exposed to 90% N₂,5% O₂,5% CO₂ for 2,6,12,and 24 hours.The apoptosis of the cells was identified by Hoechst 33258 staining and the expressions of c-jun,c-fos was analyzed with western-blot.ResultsWith the extension period of hypoxia,the number of apoptotic cells was significantly increased from 23.79±3.43% to 74.36±5.58%.Western-blot assay showed the expressions of c-jun,c-fos increased at 2 hr,and reached the highest at 6 hr and then gradually decreased,but were still higher than those of the control at 24 hr.ConclusionContinuous hypoxia can induce the expression of c-jun,c-fos in hippocampal cells in vitro,which might be related to the cell apoptosis and the neuron protective mechanism.