镍冶炼烟尘对NIH/3T3细胞bcl-2和bax蛋白表达水平的影响  

The effect of nickel-smelting fumes on the expression of bcl-2 and bax in NIH/3T3 cells

作　　者：张琳[1] 王玥[1] 马文翰[1] 付瑶[1] 韩丹[1] 吴永会[1] 

机构地区：[1]哈尔滨医科大学公共卫生学院劳动卫生与职业病学教研室,150081

出　　处：《中华劳动卫生职业病杂志》2015年第3期175-179,共5页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金资助项目（81372965）

摘　　要：目的 研究镍冶炼烟尘对NIH/3T3细胞bcl-2和bax蛋白表达水平的影响.方法 取生长对数期的NIH/3T3细胞,置于镍冶炼烟尘浓度分别为0、6.25、12.50、25.00、50.00、100.00 μg/ml的培养液中培养24 h,噻唑蓝（MTT）法检测细胞存活率、乳酸脱氢酶（LDH）试剂盒测定细胞LDH活力、Hoechst33342法观察细胞凋亡形态学改变以及蛋白免疫印迹法（western blot）检测NIH/3T3细胞bcl-2和bax蛋白表达水平.结果 与对照组比较,各时点各染毒组细胞相对存活率明显降低,除作用7d6.25 μg/ml染毒组外,差异均有统计学意义（P＜0.05）.胞外LDH活力随染毒浓度的增加而升高,各染毒组胞外LDH活力明显高于对照组,差异有统计学意义（P＜0.05）.经100.00 μg/ml镍冶炼烟尘作用24 h细胞呈现染色质浓缩、细胞体积缩小等凋亡早期形态学特征.6.25、12.50、25.00μg/ml染毒组细胞bcl-2蛋白表达分别为0.58±0.01,0.63±0.01,0.57±0.01,均高于对照组,差异有统计学意义（P＜0.05）,50.00、100.00 μg/ml染毒组细胞bcl-2蛋白表达分别为0.35±0.01,0.27±0.01,均低于对照组,差异有统计学意义（P＜0.05）.6.25μg/ml染毒组细胞bax蛋白表达量为0.58±0.00,明显低于对照组,差异有统计学意义（P＜0.05）,50.00、100.00μg/ml染毒组bax蛋白表达量分别为0.71±0.01,0.78±0.02,均高于对照组,差异有统计学意义（P＜0.05）.结论 NIH/3T3细胞暴露镍冶炼烟尘24h后能够引起细胞凋亡的启动,并且氧化应激可能诱导其凋亡的发生.Objective To investigate the effect of nickel-smelting fumes on the expression of bcl-2 and bax in mammalian cells.Methods Logarithmic growth NIH/3T3 cells were exposed to venom for 24 h,which sample fumes concentration was respectively 0,6.25,12.50,25.00,50.00,100.00 μg/ml.Cell viability was assessed by MTT assay and the level of extracellular LDH activity was detected with Lactate Dehydrogenase （LDH） kit.Morphological changes of apoptotic were observed with Hoechst33342,while Western blot was used to measure the expression of bcl-2 and bax.Results In addition to 7 days of 6.25 μg/ml nickel-smelting fumes group,each time point and dose group＇s cell viability reduced with significant differences compared with the control group （P〈0.05）.the extracellular LDH activity increased with increasing dose of nickel-smelting fumes,and the extracellular LDH activity of 6.25、12.50、25.00、50.00、100.00 μg/ml nickel-smelting fumes group increased as compared with the control group （P〈0.05）.Simultaneously,the cells,treated with 100.00 μg/ml nickel-smelting fumes for 24 h,appeared obvious morphological changes of apoptosis,such as chromatin condensation and cell shrinkage.the expression of bcl-2 significantly increased in groups of 6.25、12.50、25.00 μg/ml nickel-smelting fumes （0.58±0.01,0.63±0.01 and 0.57±0.01） and decreased in groups of 50.00、100.00 μg/m nickel-smelting fume（0.35±0.01 and 0.27±0.01） as compared with that of the control group（P〈0.05）.And the expression of bax significantly decreased in group of 6.25μg/ml nickel-smelting fumes（0.58±0.00） and increased in groups of 50.00、100.00 μg/m nickel-smelting fumes （0.71 ±0.01 and 0.78±0.02） as compared with that of the control group （P〈0.05）.Conclusion Apoptosis was activated in NIH/3T3 cell after 24 h of exposure to Ni-smelting fumes,which may be induced by oxidative stress.

关 键 词：镍 粉尘 氧化性应激 BCL-2 BAX 

分 类 号：R131[医药卫生—劳动卫生]