tBHQ对NaAsO_2诱导HaCaT细胞凋亡的拮抗作用

Antagonism of tBHQ on HaCaT cells apoptosis induced by NaAsO_2

目的探讨叔丁基对苯二酚(tertiary butylhydroquinone,tBHQ)对亚砷酸钠(NaAsO2,sodium arsenite)诱导人角质上皮HaCaT细胞凋亡的拮抗作用。方法 tBHQ(10、25、50μmol/L)预处理12 h,再与NaAsO2(5、10、30μmol/L)共同处理HaCaT细胞24 h。用Annxin V/PI染色流式细胞术法检测细胞凋亡率;DAPI染色观察细胞形态学改变;JC-1法测定线粒体膜电位。结果将实验数据进行ANOVA分析处理后表明,5,10、30μmol/L NaAsO2单独作用时,细胞凋亡率与对照组相比显著升高,而线粒体膜电位则显著下降(P<0.05或P<0.01),形态学观察可见高强度DAPI染色细胞数目增加和凋亡小体出现;当给予tBHQ(10、25、50μmol/L)预处理后,NaAsO2致HaCaT细胞凋亡率升高和线粒体膜电位下降均明显受到抑制(P<0.05),形态学观察可见高强度DAPI染色细胞数目和细胞核碎片明显减少。结论实验结果表明tBHQ可能通过线粒体凋亡途径抑制NaAsO2引起的HaCaT细胞凋亡。

Objective To discuss the antagonistic effect of tertiary butylhydroquinone（tBHQ） on human keratinocytes cells（HaCaT） apoptosis induced by sodium arsenite（NaAsO2）.Methods HaCaT cells was pretreated with tBHQ（10,25 and 50 μmol/L） for 12 hours,then was treated together with NaAsO2（5,10 and 30 μmol/L） for 24 hours.The rate of cell apoptosis was determined by Annexin V/PI staining flow cytometry;the morphology was evaluated by fluorescence microscopy following DAPI staining.Mitochondrial membrane potential was determined by JC-1 method.Result The results of ANOVA test indicated the rate of apoptosis significantly increased treated with NaAsO2（5,10 and 30 μmol/L） compared with control group and the depolarization of mitochondrial membrane potential gradually decreased compared with the control group（P0.05 or P0.01）.The morphology showed the number of staining cells was significantly increased and apoptotic body appeared in high strength DAPI staining.The increased rate of HaCat cell apoptosis and decreased mitochondrial membrane potential were inhibited in the condition of pretreating with tBHQ（10,25 and 50 μmol/L）（P0.05）.The morphology showed the number of staining cells and cell debris were significantly reduced.Conclusions The results of the study indicate that tBHQ inhibits HaCaT cells apoptosis induced by NaAsO2 by mitochondrial apoptotic pathway.