微波暴露对原代培养大鼠海马神经元热休克蛋白家族表达的影响  

Effect of microwave irradiation on expression of heat shock proteins family in primary cultured rat hippocampal neurons

作　　者：屈明玥[1] 邓朝晖[1] 周舟[1] 魏爱民[1] 陈纯海[1] 余争平[1] 王登高[1] 

机构地区：[1]第三军医大学劳动卫生学教研室,教育部电磁辐射医学防护重点实验室,重庆400038

出　　处：《中华劳动卫生职业病杂志》2010年第12期909-913,共5页Chinese Journal of Industrial Hygiene and Occupational Diseases

摘　　要：目的 研究微波暴露对原代培养大鼠海马神经元中热休克蛋白（heat shock proteins,HSPs）表达调控的影响.方法 采用平均功率密度为90 mW/cm2微波辐照成熟的原代培养大鼠海马神经元10 min,于辐照后0、3、6、12、24 h提取细胞总蛋白,采用免疫印迹（Western blot）法检测HSP27、HSP70、HSP90和热休克转录因子（heat shock factor 1,HSF1）蛋白表达情况;于辐照后0、3、6、12、24 h分别提取细胞总RNA和总蛋白,采用实时定量聚合酶链反应（PCR）检测hsf1的mRNA表达水平;于辐照后20、40 min提取细胞核蛋白,凝胶迁移滞后实验（electrophoretic mobility shift assays,EMSA）法检测HSF1的活性变化.结果 90 mW/cm2微波辐照10 min后,HSP27蛋白表达水平在辐照后3、6、12 h分别明显升高22%、36%、18%,HSP70蛋白表达水平在辐照后3、6、12 h分别明显升高23%、32%、26%,与辐照前比较,差异均有统计学意义（P＜0.05,P＜0.01）.HSP90蛋白表达水平在辐照后6、12 h分别明显升高27%、33%,与辐照前的差异亦有统计学意义（P＜0.05,P＜0.01）.HSF1 mRNA和蛋白表达水平未见明显变化,但微波辐照可激活HSF1的DNA结合活力. 结论微波辐照后大鼠海马神经元发生热休克反应,被活化的HSF1在转录水平上调控HSPs的表达,从而发挥拮抗微波损伤的作用.Objective To investigate the effects of microwave irradiation on the expression and regulation of heat shock proteins（HSPs）in primary cultured rat hippocampal neurons.Methods Neurons were exposed to 90 mW/cm2 microwave irradiation for 10 minutes.Western blot was used to determine the expression of HSP27, HSP70, HSP90 and heat shock factor 1（HSF1）at 0, 3, 6, 12 and 24 hour respectively.Realtime RT-PCR was used to determine the mRNA expression of HSF1.DNA-binding activity of HSF1 was measured by electrophoretic mobility shift assay（EMSA）.Results The protein expression of HSP27 was significantly increased by 22%, 36%, 18% at 3, 6, 12 h, respectively（P＜0.05）.The protein expression of HSP70 was significantly increased by 23%, 32%, 26% at 3, 6, 12 h, respectively（P＜0.05,P＜0.01）.The protein expression of HSP90 was significantly increased by 27%, 33% at 6, 12 h, respectively（P＜0.05 ,P＜0.01）.The DNA-binding activity of HSF1 was stimulated, however, no significant change of the expression of HSF1 was observed on both the mRNA and protein levels.Conclusion The transcriptional activity of HSF1 is activated by microwave irradiation, which promotes the expression of HSPs.Heat shock response which contributes to establish a cytoprotective state is induced by microwave irradiation in primary cultured rat hippocampal neurons.

关 键 词：微波 热休克蛋白质类 神经元 海马 

分 类 号：R730.51[医药卫生—肿瘤]