重复高压氧预处理对大鼠皮质神经元缺氧损伤的影响

Effect of repeated hyperbaric oxygen preconditioning on hypoxic injury in rat cortical neurons

目的观察重复高压氧预处理对大鼠皮质神经元缺氧损伤的影响。方法将对数生长期皮质神经元分为四组:正常对照组(C组),细胞置常规培养箱内培养;高压氧预处理组(P组),培养细胞行0.35 MPa、2 h高压氧预处理,1次/d,连续3 d;缺氧处理组(H组),细胞置三气培养箱内培养4 h,设置培养条件为95%N2、5%CO2;高压氧预处理加缺氧处理组(PH组),高压氧预处理后行缺氧处理,条件设置同上。四组细胞均在缺氧处理4 h后行MTT和彗星(单细胞凝胶电泳)试验。结果 C组与H组之间细胞活力和尾矩的差异有统计学意义(P<0.05);与H组相比,PH组细胞活力明显增高(P<0.05),尾矩明显降低(P<0.05)。结论重复高压氧预处理提高皮质神经元活力,降低细胞的DNA损伤程度,对缺氧损伤的皮质神经元具有保护作用。

Objective To investigate the effect of repeated hyperbaric oxygen preconditioning on hypoxic injury in primary cultured cortical neurons. Methods Primary cultured rat cortical neurons were randomly assigned to 4 groups, i. e. control group （ Group C ） generally cultured in eommon incubator, hyperbaric oxygen preconditioning group （ Group P） preconditioned with hyperbaric oxygen （in proposal of 0.35 MP, 2 h, once/d, 3 d consecutively）, hypoxia group （Group H） cultured in three gas incubator （3 h, 95% N2 , 5% CO2 ） , and hyperbaric oxygen preconditioning ＋ hypoxia group （ Group PH） cultured in three gas incubator after precondi- tioning with hyperbaric oxygen. Methyl thiazolyl tetrazolium （MTT） assay and comet assay （single cell gel electrophoresis） were performed after 4 h in hypoxia culture. Results The differences in cytoactivity and tail moment were statistical significance between Group C and Group H（ P 〈 0.05 ） ; compare with that of Group H, the cytoactivity of Group PH increased obviously （P 〈0.05 ）, and the tail moment decreased obviously （ P 〈 0.05 ）. Conclusion Hyperbaric oxygen preconditioning improves the cytoactivity, relieves the DNA damage of cortical neurons, and induces the tolerance against hypoxia injury in cortical neurons.