缺血预适应对大鼠海马神经元凋亡的影响

目的 探讨缺血预适应对缺氧复氧后大鼠海马神经元的影响.方法 用300μmol/L氯化钴预处理大鼠海马神经元2 h,然后更换正常的培养基培养24 h,之后用无血清的培养基培养,建立预适应的细胞模型.四甲基偶氮唑蓝(MTT)法测定细胞增殖,流式细胞术测定细胞凋亡,反转录-聚合酶链反应(RT-PCR)检测凋亡相关基因的表达情...

Protective effects of ischemic preconditioning on anoxia-reoxygenation induced apoptosis in rat hippocampal neurons

Objective To establish a cell preconditioning model and clarify the regulation of ERK1/2 and apoptosis related gene in ischemic preconditioning.Methods Newborn SD rats were decapitated,hippocampal tissue was isolated and hippocampal neurons were prepared by digestion with trypsin.After being cultured,the hippocampal neurons were randomly assigned into 3 groups.The neurons were pretreated with 300 μmol/L CoCl2 for 2 hours,and then they were stimulated with no serum media 24 hours later.MTT method was used to detect the proliferation of cell.Then RT-PCR method was used to show the regulation of apoptosis related gene in ischemic preconditioning.Western blot method was used to find the change of ERK1/2.Results After pretreated with 300 μmol/L CoCl2 for 2 hours,cell proliferation increased when cultured with no serum media 24 hours later,bcl-2 gene was up-regulated and caspase-3 gene was down-regulated(P<0.05).Meanwhile,ERK1/2 was down-regulated after preconditioning(P<0.05).Conclusion Pretreating the neurons with 300 μmol/L CoCl2 can protect rat hippocampal neurons from anoxia-reoxygenation induced apoptosis by inhibiting the expression of ERK1/2.