三(2-氯乙基)磷酸酯诱导肝细胞线粒体毒性的研究  

Tris(2-chloroethyl)phosphate Induced Mitochondrial Toxicity in Hepatocytes

作　　者：张文娟[1] 张佑健 王志远[1] 殷文军[1] 徐甜[1] 郑红燕[1] 熊伟[1] 袁晶[1] 

机构地区：[1]华中科技大学同济医学院公共卫生学院劳动卫生与环境卫生学系环境与健康教育部重点实验室国家环境保护环境与健康重点实验室(武汉)省部共建国家重点实验室培育基地-湖北省环境卫生学重点实验室,湖北430030

出　　处：《环境与职业医学》2015年第10期931-935,940,共6页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金资助项目(编号:81273023)

摘　　要：[目的]研究三(2-氯乙基)磷酸酯[tris(2-chloroethyl)phosphate,TCEP]对肝细胞线粒体功能的影响。[方法]用0.00(溶剂对照组)、3.12、12.50、50.00和200.00 mg/L TCEP分别处理人正常肝细胞(L02细胞)和人肝癌细胞(Hep G2细胞)24 h和48 h。测定细胞活力、线粒体内活性氧水平、线粒体DNA拷贝数、线粒体膜电位和胞内游离钙(Ca2+)水平以及胞内ATP的浓度。[结果]与相应溶剂对照组相比,在24 h,200.00 mg/L TCEP处理组L02细胞活力降低(P<0.05),≥12.50 mg/L TCEP处理组Hep G2细胞活力降低(P<0.05);在48 h,50.00 mg/L和200.00 mg/L TCEP处理组两种细胞活力降低(P<0.05)。所有TCEP处理组两种细胞的线粒体DNA拷贝数均减少(P<0.05或P<0.01)。在24 h,所有TCEP处理组两种细胞的线粒体膜电位均下降(P<0.05或P<0.01)。在24 h,200.00 mg/L TCEP处理组两种细胞胞内游离Ca2+水平均升高(P<0.01);在48 h,50.00 mg/L和200.00 mg/L TCEP处理组两种细胞胞内游离Ca2+均升高(P<0.01)。在24 h和48 h,200.00 mg/L TCEP处理组L02胞内ATP水平降低(P<0.05);在24 h,50.00 mg/L和200.00 mg/L TCEP处理组Hep G2胞内ATP浓度下降(P<0.05或P<0.01)。[结论]一定浓度的TCEP可致肝细胞线粒体损伤和线粒体功能指标异常,提示TCEP有肝细胞线粒体毒性。[ Objective ] To evaluate effects of tris（2-chloroethyl）phosphate （TCEP） on mitochondrial functions in liver cells. [ Methods ] At 24 and 48h after L02 and HepG2 cells were treated with 0.00 （solvent control group）, 3.12, 12.50, 50.00, 200.OOmg/L TCEP, we detected cell viability, mitochondrial reactive oxygen species （mtROS） levels, mitochondrial DNA copy numbrane, mitochondrial membrane potential, intracellular free Ca2＋ levels, and intracellular ATP level. [ Results ] Compared with the control group, TCEP decreased the cell viabilities of L02 cells in the 200 mg/L TCEP group （P 〈 0.05） and the cell viabilities of HepG2 cells in the ≥ 12.50mg/L TCEP groups （P〈0.05） at 24h; decreased the cell viabilities of L02 and HepG2 cells in the 50.00 and 200.00 mg/L TCEP groups at 48 h （P 〈 0.05）; reduced the mitochondrial DNA number of L02 and HepG2 cells in all treatment groups （P 〈 0.05 or P 〈 0.01）; decreased the mitochondrial membrane potential of L02 or HepG2 cells in all treatment groups at 24 h （P〈 0.05 or P 〈 0.01）; increased the intracellular free Ca2＋ concentrations of L02 and HepG2 cells only in the 200.00mg/L TCEP group at 24h and in the 50.00 and 200.00 mg/L groups at 48 h （P 〈 0.01）; decreased the intracellular ATP levels of L02 cells only in the 200.00 mg/L TCEP group at 24 and 48 h （P 〈 0.05） and in the 50.00 and 200.00 mg/L TCEP groups of HepG2 cells at 24 and 48 h （P 〈 0.05 or P 〈 0.01）. [ Conclusion ] A certain concentrations of TCEP could cause mitochondrial damage and abnormal changes in indices reflecting mitochondrial dysfunctions, implying that TCEP could induce mitochondrial toxicity in hepatocytes.

关 键 词：三(2-氯乙基)磷酸酯 线粒体 ATP 线粒体膜电位 胞内游离钙离子 活性氧 

分 类 号：R114[医药卫生—卫生毒理学]