三氯乙烯和四氯乙烯对体外培养皮肤KC细胞氧化损伤及维生素E的保护作用

Effects of TCE and PCE on cultured human keratinocyte lipid peroxidation and protective effect of Vitamin E on it

目的探讨三氯乙烯(TCE)、四氯乙烯(PCE)对体外培养人皮肤角质形成细胞(KC)氧化损伤作用及维生素E的保护作用。方法将来源于3个或3个以上不同健康个体的皮肤KC细胞混合培养于K-SFM培养基中,利用中性红吸附实验确定其半数抑制浓度(IC50),并据此确定TCE、PCE染毒浓度。染毒4 h后,测定细胞内丙二醛(MDA)、超氧化物歧化酶(SOD)、活性氧(ROS)水平;维生素E保护组使用不同浓度维生素E预作用2 h后,再分别换入0．5 mmol/L TCE或0．2 mmol／L PCE与不同浓度维生素E混合物作用4 h后,测定细胞内MDA、SOD、R0s水平。结果TCE、PCE能引起体外培养人皮肤KC细胞氧化损伤,且具有浓度-反应关系。而维生素E对其具有保护作用,能下调其氧化损伤,且具有浓度-反应关系。结论TCE、PCE能引起体外培养人皮肤KC细胞氧化损伤,而维生素E对其具有保护作用。

Objective To explore the different concentrations of trichloroethylene （TCE） or perchloroethylene （PCE） induced cultured normal human epidermal keratinocyte （KC） lipid peroxidation and protective effect of Vitamin E on it. Methods KC derived from 3 or more donors were pooled together and cultured with K-SFM. Neutral Red Uptake Assay was used to determine the IC50 of TCE or PCE, and then different concentrations of TCE or PCE were administered for culturing KC; 0.5mmol/L TCE or 0.2mmol/L PCE and different concentrations of Vitamin E were used to determine the protective effect of Vitamin E. After 4 hours＇ culture, kits were used to determine cellular MDA,SOD and ROS level. Results Treatment of KC with different concentrations of TCE or PCE showed significant dose-related variations in lipid peroxidation,with the higher concentration, higher level of MDA, ROS and lower activity of SOD displayed in this study. Vitamin E 10 - 200 mmol/L dose-dependently attenuated MDA and ROS level, and increased SOD activities. Conclution TCE or PCE can induce the lipid peroxidation in cultured KC and Vitamin E protects it from TCE- or PCE-induced peroxidation.