镍冶炼烟尘染毒致NIH／3T3细胞线粒体损伤及L-抗坏血酸的干预作用  

Nickel smelting dust exposures to NIH/3T3 cellular mitochondrial damage and L-ascorbic acid interference effect

作　　者：潘昱霖 王玥[1] 付瑶[1] 王声远[1] 谭文乔 陈嘉述 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》2016年第3期173-177,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小鼠胚胎成纤维细胞（NIH／3T3）线粒体损伤的影响及L-抗坏血酸的保护作用。方法将NIH／3T3细胞分为染毒组和L-抗坏血酸干预组。染毒组加入镍冶炼烟尘配制浓度分别为0.00、25．00、50.00、100．00g/ml的混悬液，干预组在染毒前额外加入L-抗坏血酸（100mmol／L）。利用MTT法检测细胞生长抑制率，钙黄绿素-AM荧光探针检测细胞线粒体膜通道孔（MPTP）变化，JC-1荧光染色观察并检测细胞线粒体膜电位（MMP）变化，比色法定量检测细胞线粒体呼吸链复合物Ⅰ、Ⅱ、Ⅲ、Ⅳ活性变化。结果随着染毒浓度升高，染毒组和干预组细胞生长抑制率均明显增加。在25．00、50．00、100．00μg/ml浓度时干预组抑制率均低于染毒组，差异有统计学意义（P〈0．05）。在50．00、100．00μg/ml浓度时，染毒组MPTP荧光强度为564．940±14．015、436．613±31．275；干预组MPTP荧光强度为611．203±20．053、498．840±21．308，干预组的荧光强度均高于染毒组，差异有统计学意义（P〈0．05）；荧光显微镜下干预组绿色荧光强度低于染毒组，干预组红色荧光强度高于染毒组。在25．00、50．00、100．00±g／ml浓度时，染毒组MMP绿色与红色荧光比值分别为16．579±0．220、14．453±0．280、11．133±1.007；干预组MMP绿色与红色荧光比值分别为18．697±0．163、16．260±0．520、13．703±0．715，干预组绿色与红色荧光比值均高于染毒组，差异有统计学意义（P〈0．05）。在25．00、50．00、100．00μg／ml浓度时，干预组的复合物Ⅰ、Ⅳ活性均高于染毒组，在100μg／ml浓度时，复合物Ⅱ染毒组活性为0．754±0．013，干预组活性为0．827±0．032，干预组的复合物Ⅱ活性高于染毒组，差异有统计学意义（P〈0．05）。结论随着镍冶炼烟尘浓度升高，NIH/3T3细胞线粒体损伤程度上升，L-抗坏血酸对镍冶炼烟尘�Objective To study the protection of L-ascorbic acid on mouse embryonic fibroblasts（NIH/ 3T3） from carcinogenic effects caused by nickel smelting smoke subjects. Methods The NIH/3T3 cells were divided into experimental and L-ascorbic acid in the intervention group. Plus exposure group concentration of nickel refining dusts were formulate 0.00, 25.00, 50.00, 100.00 μg/ml suspension, the intervention group on the basis of the added exposure group containing L-ascorbic acid （100 mmol/L）, contacted. Then, the cell viability was detected by MTT assay, we used Calcein-AM fluorescence probe to detect cell mitoehondrial permeability transition pore （MPTP） changes, JC-1 staining to observe and detect the cell mitochondrial membrane potential （MMP） change, colorimetric quantitative to study the activity of mitochondrial respiratory chain complex Ⅰ, Ⅱ,Ⅲ, Ⅳ. Results Upon 24 h incubation, both cell relative inhibition rate, openness of MPTP were increasing enhanced by different concentrations, on the other hand, MMP and the activity of mitoehondrial respiratory chain complex Ⅰ, Ⅱ, Ⅳ were obviously decreased, the differences were statistically significant （P〈0.05）.After L-ascorbic acid intervention treatment, the results of the intervention group were lower than that of the exposure group, and the difference was statistically significant （P〈0.05）, indicating the protection of L-aseorbic acid on cell mitochondrial from the nickel exposure damage. Conclusion The damage effects of nickel on NIH/3T3 cell mitochondrial was significantly enhanced with the increasing concentration, and L-ascorbie acid has certain protection on cellular mitochondrial.

关 键 词：镍 冶炼烟尘 线粒体 L-抗坏血酸 

分 类 号：R135[医药卫生—劳动卫生]