电磁脉冲辐照对培养大鼠海马神经元细胞内培养上清LDH及培养上清液中CHE、K^+、Na^+浓度的影响

EMP affects the LDH concentration inner neurons of rat 's hippocampus and LDH、CHE、K~ +、Na^+ in culture medium

目的:利用测定电磁脉冲辐照原代培养海马脑神经细胞前后细胞内LDH和培养上清液中LDH、CHE、K+、Na+浓度及与时间的关系,探讨电磁脉冲对海马神经细胞损伤的机制。材料和方法:将新生的Wistar(24小时内)乳鼠断头,解剖镜下剔除脑膜、血管,分离出海马组织,将消化洗涤后的海马神经细胞稀释成1×106个/ml浓度的细胞悬液;然后以2ml/皿的量,将细胞悬液种植到35mm培养皿中,放入37℃、5%CO2-95%空气条件下培养。在培养12～14天时,高场强EMP模拟源(有界波模拟源),场强为6×104V/m,脉冲上升时间为20ns,脉宽为30μs,频率包含0～100MHz。以2.5次/分,辐照2分钟。并于辐照后0h(即刻)、1h、6h、12h和24h应用中生公司生化检测试剂盒测定细胞内和培养上清中LDH及培养上清液中CHE、K+、Na+浓度。结果:电磁脉冲辐照后即刻就可引起培养上清LDH明显升高;辐照后1h细胞内LDH明显降低,而培养上清中LDH、CHE、和K+明显升高;辐照后6h细胞内LDH明显降低,而培养上清中LDH、CHE、K+和Na+明显升高;辐照后12h细胞内LDH明显降低,培养上清中CHE、K+和Na+明显升高;辐照后24h所有上述指标基本恢复。结论:电磁脉冲辐照后可损伤大鼠海马神经元细胞膜,表现为膜通透性升高或膜穿孔。

Objective:With assaying the changes of LDH concentration in rat's hippocampus neurons and LDH、CHE、K^+ 、Na^+ in culture medium after irradiating by electromagnetic pulse(EMP)to explore its injury mechanism to neurons.Methods:Hippocam-pus neurons come from neonatal rat were cultured in6-well culture plates.One plate was control sample.The all five plates were irradiated by high field strength5EMP within2minutes,The electromagnetic pulse simulator which provides5pulses /min with a high electric field intensity60KV/m,20-nsec rise time and30μs pulse wide.The LDH concentration in neurons and LDH,AST,CHE,K + and Na + in culture medium were measured at0h,1h,6h,12h and24h following the cells being ir-radiated,and were measured by reagent kits(Beijing Zhongsheng high-tech bioengineering company).All data was analyzed by statistical software Spss8.0.Results:LDH in culture medium was increased significantly at0h after irradiated by EMP (P<0.05).LDH in neurons was decreased significantly,but LDH,CHE and K + in culture medium at1h were increased after irradiated by EMP (P<0.01).LDH in neurons was decreased significantly,but LDH,CHE,K + and Na + in culture medium at6h were in-creased after irradiated by EMP (P<0.05).LDH in neurons was decreased significantly,but increased CHE?K + and Na + in culture medium at12h after irradiated by EMP (P<0.05).All above items tend to recover to control plate.Conclusion:EMP could in-jured the membranes of hippocampus neurons.