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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 investigate the antagonistic effect of tertiary butylhydroquinone (tBHQ) on the apoptosis of human keratinized HaCaT cells induced by sodium arsenite (NaAsO2). Methods HaCaT cells were treated with tBHQ (10, 25 and 50 μmol / L) for 12 h and HaCaT cells were treated with NaAsO2 (5, 10 and 30 μmol / L) for 24 h. The apoptosis rate was detected by flow cytometry with Annxin V / PI staining. The morphological changes of cells were observed by DAPI staining. The mitochondrial membrane potential was measured by JC-1 method. Results The experimental data were analyzed by ANOVA. The apoptotic rate of 5,10,30μmol / L NaAsO2 alone was significantly higher than that of the control group, while the mitochondrial membrane potential was significantly decreased (P <0.05 or P < 0.01). The morphological observation showed that the number of high-intensity DAPI-stained cells increased and apoptotic bodies appeared. After pretreatment with tBHQ (10,25,50μmol / L), the apoptotic rate of HaCaT cells induced by NaAsO2 and mitochondrial membrane potential (P <0.05). Morphological observation showed that the number of DAPI-stained cells and the number of nuclear fragments were significantly decreased. Conclusion The experimental results show that tBHQ may inhibit NaAsO2-induced apoptosis of HaCaT cells through the mitochondrial apoptotic pathway.