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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。
Objective To investigate the antagonistic effect of tert-butylhydroquinone (tBHQ) on the cell cycle arrest of HaCaT cells and the abnormal changes of its regulatory proteins induced by sodium arsenite (NaAsO2). Methods The cell cycle was determined by flow cytometry. The protein expression of Cyclin D1 and CDK4 was detected by Western Blot. Results Compared with the control group, the percentage of cells in S phase and G2 / M phase was significantly increased (P <0.01), and the ratio of tBHQ (50μmol / (P <0.01). The proportion of S phase cells in NaAsO2 (50μmol / L) pretreatment group treated with tBHQ (50μmol / L) (P <0.01). The proportion of cells in G2 / M phase showed a decreasing trend as a whole. The protein expression of cyclin D1 and CDK4 in NaCaT cells treated with NaAsO2 decreased significantly with the increase of NaAsO2 concentration (P <0.01). After pretreatment with tBHQ, the protein expression of Cyclin D1 and CDK4 were significantly higher than that of NaAsO2 Concentration of NaAsO2 alone group (P <0.01). Conclusion tBHQ can antagonize the abnormal cell cycle of human skin keratinocytes induced by inorganic arsenic.