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[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。
[Objective] To investigate the effect of di (-2-ethylhexyl) phthalate (DEHP) and benzo (a) pyrene [benzo (HepG2 cells) on their cytochrome 1A1 (CYP1A1) and cytochrome P450 (3A4) CYP3A4 activities. [Methods] HepG2 cells were treated with B (a) P 64.0 μmol / L and DEHP 62.5, 125.0, 250.0, 500.0 and 1000.0 μmol / L respectively for 48 h and 72 h. Solvent control group was dimethyl sulfoxide (DMSO <1 ‰). Cell viability was measured by CCK8 assay; CYP1A1 and CYP1A1 were evaluated using 7-ethoxyisoxypyoxazole-O-deethoxylase and 7-ethoxycoumarin O-deethylase (EROD and ECOD) CYP3A4 activity. The mRNA and protein expression levels of CYP1A1 and CYP3A4 were detected by real-time fluorescence quantitative PCR and Western blot respectively. [Results] Compared with DEHP alone group, the survival rate of cells in combined exposure group was significantly decreased (P <0.01), while the activities of EROD and ECOD were significantly increased (P <0.05, P <0.01). Combined treatment with CYP1A1 Only the mRNA expression of CYP3A4 mRNA and protein were significantly increased compared with DEHP alone group (P <0.01). [Conclusion] The combination of DEHP and B (a) P induced the activity of CYP1A1 and CYP3A4 in HepG2 cells, and had some effects on the expression of CYP1A1 and CYP3A4 at mRNA and protein levels.