三氯乙烯对人原代角质形成细胞的细胞毒作用  

Cytotoxicity of Trichloroethylene on Normal Human Epidermal Keratinocytes

作　　者：沈彤[1] 丁锐[1] 涂登云[1] 马泰[1] 周承藩[1] 于均峰[1] 朱启星[1] 

机构地区：[1]安徽医科大学公共卫生学院,安徽合肥230032

出　　处：《工业卫生与职业病》2006年第1期24-28,共5页Industrial Health and Occupational Diseases

基　　金：国家自然科学基金(30471469);安徽省自然科学基金项目(03043801);安徽省教育厅自然科学重点科研项目(2003KJ036ZD)资助

摘　　要：目的研究体外培养条件下有机溶剂三氯乙烯（Trichloroethylene，TCE）对正常人表皮角质形成细胞（normal humanepidermal keratinocyte，NHEK）的毒性作用。方法用0．25％的胰蛋白酶经冷温两步消化皮肤，分离得到NHEK．进行体外无血清培养；根据中性红吸附试验测定的NR50结果，确定TCE染毒剂量。测定乳酸脱氢酶（LDH）活力，反映其对细胞膜的损害，测定丙二醛（MDA）含量和超氧化物歧化酶（SOD）活力．反映细胞脂质过氧化作用。结果TCE可引起NHEK细胞活力剂量依赖性降低，TCE对NHEK的NR50值为4．525mmol／L（3．922～5．128mmol／L）；NHEK细胞用0．125，0．250，0．500，1．000，2．000mmol／L的TCE处理1，2，3，4h后，LDH的释放显示明显的时间-剂量-反应关系；同样剂量的TCE处理NHEK，4h后可引起MDA含量呈浓度依赖性增加（最低浓度为0．500mmol／L）．SOD活力呈浓度依赖性抑制（最低浓度为0．250mmol／L），与空白对照和溶剂对照组相比较，两者差异均有显著性（P〈0．05）。结论在体外培养条件下，有机溶剂三氯乙烯可通过脂质过氧化和氧化应激作用对人角质形成细胞产生明显的细胞毒作用。Objective To study the cytotoxieity of trichloroethylent （TCE） on normal human epidermal keratinoeytes （NHEK） in vitro. Methods Administered doses of TCE to NHEK were determined according to neutral red uptake （NRU） assay with free serum culture in vitro. Membrane damage and lipid peroxidation （LPO） were assessed by lactate dehydrogenase （LDH） release test and measurement of malondialdehyde （MDA） contents and superoxide dismutase （SOD） activity. Results TCE could induce a dose-dependent decrease in cell viability, NR50 values of TCE on NHEK was found to be 4. 525 mmol/L （CI 95% ： 3. 922- 5. 128）. A time-and concentration-dependent release of LDH was observed at 1, 2, 3, and 4 h after cells were exposed to 0. 125, 0. 250, 0. 500, 1. 000, 2. 000 mmol/L TCE. When the same concentration TCE treated NHEK for 4h, TCE also induced an increase of MDA content, whilst an inhibition of SOD activity in a concentration-dependent manner. The lowest concentrations compared with the blank and vehicle control groups were 0. 500 and 0. 250 mmol/L, respectively. Conclusions These results suggest that TCE could induce cytotoxicity to cultured NHEK in vitro, which was associated with lipid peroxidation and oxidative stress.

关 键 词：三氯乙烯 角质形成细胞 细胞毒作用 

分 类 号：R114[医药卫生—卫生毒理学]