叔丁基对苯二酚对人正常肝细胞的毒性

Cytotoxicity of tertiary butylhydroquinone to human normal hepatic cells

目的:研究叔丁基对苯二酚在大鼠体内代谢后的浓度及在该浓度范围内的体外细胞毒性。方法:大鼠高剂量(700mg/kg)灌胃后,运用液相色谱/质谱联用法检测血清中叔丁基对苯二酚的浓度。以人正常肝细胞HL7702为研究对象,采用MTT法检测叔丁基对苯二酚的细胞毒性,采用倒置显微镜观察细胞形态变化,并评估叔丁基对苯二酚在体内浓度范围内的体外细胞毒性。结果:叔丁基对苯二酚抑制细胞生长,并使细胞形态学特征改变;随着浓度升高(0.01～1.2mmol/L)和细胞培养时间延长(12～48h),其对细胞的毒性作用加剧(细胞毒性等级0～4);细胞培养时间为12、24、48h时,半致死浓度分别为0.63、0.34、0.16mmol/L;大鼠血清中叔丁基对苯二酚的浓度≤0.12mmol/L,细胞毒性等级0～2级。结论:经口进入体内的叔丁基对苯二酚,代谢后的体内浓度不会引起高细胞毒性反应。

Purpose：To investigate the concentration level of tertiary butylhydroquinone in rats after oral exposure,and then to evaluate the cytotoxicity of tertiary butylhydroquinone in the concentration range observed above.Methods：After a single gavage dose of 700 mg/kg,concentrations of tertiary butylhydroquinone in rat serum were quantified by liquid chromatography/mass spectrometry（LC/MS）.Using MTT method,effects of tertiary butylhydroquinone on inhibiting proliferation of human normal hepatic cells HL7702 were studied,and changes of cell morpha were observed with inverted microscope.And then the cytotoxicity of tertiary butylhydroquinone in the concentration range in rat serum observed in vivo was evaluated.Results：Tertiary butylhydroquinone had inhibitory effect on the in vitro growth of HL7702 cells and changed the cell morpha.With the concentration of tertiary butylhydroquinone increasing from 0.01 mmol/L to 1.20 mmol/L,with the culture time increasing from 12 h to 24 h,the cytotoxicity grade increased from 0 to 4.When the culture time was 12,24,48 h,the LC50 values were respectively 0.63,0.34,0.16 mmol/L.After oral administration,concentrations of tertiary butylhydroquinone in rat serum were less than 0.12 mmol/L.Accordingly,the cytotoxicity grade in vitro was in the range of 0~2.Conclusions：Following oral exposure,the concentration level of tertiary butylhydroquinone in vivo can not result in high cytotoxicity in vitro.