B(a)P和DEHP联合染毒对HepG2细胞CYP1A1和CYP3A4酶活性的影响  

Effects of Co-exposure of Di-(2-ethylhexyl)Phthalate and Benzo(a)pyrene on CYP1A1 and CYP3A4 Enzyme Activities in HepG2 Cells

作　　者：陈曦[1] 王建书[1] 姜英[1] 杨光涛[1] 王晶[1] 汪倩[1] 饶凯敏[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,湖北武汉430030

出　　处：《环境与职业医学》2011年第9期525-530,共6页Journal of Environmental and Occupational Medicine

基　　金：国家自然科学基金面上项目(编号:30972437)

摘　　要：[目的]研究邻苯二甲酸二乙基己基酯[di(-2-ethylhexyl)phthalate,DEHP]和苯并(a)芘[benzo(a)pyrene,B(a)P]联合染毒人肝癌细胞株(HepG2细胞)对其细胞色素P450酶系1A1(cytochrome 1A1,CYP1A1)和细胞色素P450酶系3A4(CYP3A4)酶活性的影响。[方法]分别用B(a)P 64.0μmol/L和DEHP 62.5、125.0、250.0、500.0、1000.0μmol/L单独染毒和两者联合染毒HepG2细胞48h和72h。溶剂对照组为二甲基亚砜(dimethyl sulfoxide,DMSO<1‰)。用CCK8法检测细胞活性;用7-乙氧基异吩噁唑-O-去乙氧基酶和7-乙氧基香豆素O-去乙基酶(EROD和ECOD)实验评价细胞CYP1A1和CYP3A4酶活性;分别用实时荧光定量PCR法和Western blot法检测CYP1A1和CYP3A4基因的mRNA和蛋白质表达水平。[结果]与DEHP单独染毒组相比,联合染毒组细胞的存活率明显下降(P<0.01);EROD和ECOD活性却明显增加(P<0.05,P<0.01);联合染毒组CYP1A1基因仅有mRNA表达增加,但CYP3A4基因在mRNA和蛋白质表达水平均较DEHP单独染毒组明显升高(P<0.01)。[结论]DEHP和B(a)P联合染毒诱导了HepG2细胞CYP1A1和CYP3A4酶活性,并在mRNA和蛋白质水平对CYP1A1和CYP3A4的表达量产生一定影响。Effects of co-exposure of Di-（2-ethylhexyl）phthalate and Benzo（a）pyrene on CYP1A1 and CYP3A4 enzyme activities were investigated in HepG2 cells. [ Methods ] HepG2 cells were treated with either 64.0 μmol/L B（a）P alone or DEHP alone（62.5, 125.0, 250.0, 500.0, 1 000.0 μmol/L） or DEHP at the indicated concentrations plus 64.0 μmol/L B（a）P or dimethyl sulfoxide （DMSO, solvent control, 〈1‰ ） for 48 h and 72 h. The cell proliferation was evaluated by CCK8 assay. The activities of CYP 1A-associated deethylation of ethoxyresonffin （EROD） and CYP3A-associated ethoxycoumarin-O-deethylation （ECOD） were measured. Expressions of CYP1A1 and CYP3A4 at mRNA and protein were detected by quantitative real time PCR method and Western blot, respectively. [ Results ] The cell viability was decreased significantly in all co-exposure of B（a）P and DEHP groups compared with the controls （P〈0.01）. However, the EROD and ECOD activities significantly increased, when compared with the DEHP alone treated group （P 〈0.05, P 〈0.01）. Only up-regualtion of CYP1A1 transcriptional levels were observed in all co-exposure of B（a）P and DEHP groups. However, the levels of mRNA and protein expression of CYP3A4 were increased （P〈0.01）, when compared with the DEHP alone treated group. [ Conclusion ] Co-exposure of DEHP at certain concentrations and B（a）P （64.0μmol/L） induced the increases in the EROD and ECOD activities, as well as the CYP1A1 transcriptional level and the CYP3A4 transcriptional and protein levels.

关 键 词：邻苯二甲酸二乙基己基酯 苯并(a)芘 7-乙氧基异吩噁唑-O-去乙氧基酶 7-乙氧基香豆素O-去乙基酶 

分 类 号：R114[医药卫生—卫生毒理学]