【摘要】 目的 :研究PC12细胞化学性缺氧 复氧以及缺氧诱导因子 1α(HIF1α)反义寡核苷酸对HIF1α蛋白表达和氧自由基生成的影响。　方法 :制造化学缺氧 复氧模型 ,用含或不含氯化钴的DMEM培养液孵育PC12细胞。封闭HIF1α基因表达 ,用含HIF1α反义寡核苷酸和氯化钴的DMEM培养液孵育PC12细胞。HIF1α寡核苷酸进行同样处理和只用含氯化钴的DMEM培养液孵育PC12细胞作为对照。采用免疫细胞化学染色法检测HIF1α蛋白表达 ,并用激光共聚焦显微镜进行半定量。检测细胞超氧化物歧化酶 (SOD)的活性和丙二醛 (MDA)的含量 ,间接反映氧自由基的生成量。　结果 :与缺氧组相比 ,缺氧 复氧组复氧后HIF1α蛋白表达受到抑制 ,同时氧自由基生成增加。HIF1α反义寡核苷酸亦可抑制HIF1α蛋白表达和升高氧自由基水平。　结论 :抑制HIF1α的表达可导致氧自由基大量生成 ,加重缺氧 复氧PC12细胞的损伤。更多还原

【Abstract】 Objective:To antisense oligonucleotides on hypoxia inducible factor 1α(HIF1α) protein expression and oxygen free radicals production in pheochromacytoma(PC12) cells. Methods:Hypoxia-reoxygenation was induced by incubation of cells with or without cobalt chloride. Block of HIF 1α gene expression was done by incubation of cells with antisense oligonucleotides of HIF1α and cobalt chloride. Cells treated with oligonucleotides of HIF1α in the same way or incubated with cobalt chloride were considered as the control. Expression of HIF1α protein was determined by immunocytochemical staining and semi-quantified by confocal microscopy. Oxygen free radicals were represented by measurement of superoxide dismutase(SOD) activity and malonaldehyde(MDA) content. Results:HIF1α protein expression after reoxygenation in hypoxia-reoxygenation PC12 cells was reduced, whereas oxygen free radicals were increased simultaneously compared with those in hypoxic PC12 cells. Antisense oligonucleotides of HIF1α(5 μmol/L) inhibited markedly expression of HIF1α protein and increased levels of oxygen free radicals. Conclusion:Inhibition of HIF1 expression up-regulated in accumulation of oxygen free radicals and aggravation in injury of chemical hypoxia induced PC12 cells.更多还原