tBHQ对无机砷致HaCaT细胞细胞周期调节失控的拮抗作用

目的探讨叔丁基对苯二酚(tert-butylhydroquinone,tBHQ)对亚砷酸钠(sodium arsenite,NaAsO2)致人皮肤角质细胞系HaCaT细胞周期阻滞及其调控蛋白异常改变的拮抗作用。方法流式细胞仪法测定细胞周期;Western Blot检测细胞内Cyclin D1和CDK4的蛋白表达情况。结果 NaAsO2单独作用HaCaT细胞,G0/G1期细胞比例明显下降(P<0.01),S期和G2/M期细胞比例较对照组明显增加(P<0.01);tBHQ预处理后,tBHQ(50μmol/L)预处理的NaAsO2(25、50μmol/L)组细胞G0/G1期细胞比例有所上升(P<0.01);tBHQ(50μmol/L)预处理的NaAsO2(50μmol/L)组S期细胞比例明显下降(P<0.01);G2/M期细胞比例整体呈下降趋势。NaAsO2单独作用于HaCaT细胞,细胞周期相关蛋白Cyclin D1和CDK4的蛋白表达随NaAsO2染毒浓度的增加而明显下降(P<0.01);tBHQ预处理后,Cyclin D1和CDK4蛋白表达均明显高于相同浓度NaAsO2单独作用组(P<0.01)。结论 tBHQ对无机砷致人皮肤角质细胞细胞周期异常变化具有一定的拮抗作用。

Antagonistic effects of tBHQ on abnormal changes of cell cycle in HaCaT cells induced by NaAsO2

Objective To study antagonistic effects of tert-butylhydroquinone on abnormal changes of the cell cycle arrest and regulatory proteins in HaCaT cells induced by NaAsO2.Methods The flow cytometry method for the determination of the cell cycle;Western blot analysis detected the protein expression levels of Cyclin D1 and CDK4.Results HaCaT cells were exposed to NaAsO2,the proportion of cells in G0/G1 phase decreased significantly(P<0.01),the proportion of cells in S and G2/M phase rose clearly compared with the control group(P<0.01).The NaAsO2(25,50 μmol / L) group pretreated by tBHQ(50 μmol / L) in G0/G1 phase increased(P<0.01),NaAsO2(50 μmol / L) group pretreated by tBHQ(50 μmol / L) in S phase were decreased(P<0.01),cell proportion in G2 / M phase overall showed a downward trend.HaCaT cells were exposed to NaAsO2,cell cycle related proteins Cyclin D1 and CDK4 protein expression significantly decreased with the increasing dose of NaAsO2(P<0.01).Cyclin D1 and CDK4 proteins expression were significantly higher than the expression of the same concentration of NaAsO2 group(P<0.01) after being pretreated by tBHQ.Conclusion tBHQ has antagonistic effects on abnormal cell cycle changes in HaCaT cells induced by NaAsO2.