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目的探讨叔丁基对苯二酚(tBHQ)对亚砷酸钠(NaAsO2)致人皮肤角质细胞系HaCaT细胞损伤的保护作用。方法用Alamar Blue还原法检测细胞活力,用荧光探针2’,7’-二乙酰二氯荧光素(DCFH-DA)检测细胞内活性氧(ROS)的生成,用硫代巴比妥酸(TBA)法检测细胞内过氧化脂质降解产物丙二醛(MDA)的生成。结果NaAsO2(25、50μmol/L)单独处理后,各组细胞活力明显下降(P<0.01),分别为82%和59%;细胞经tBHQ(25、50μmol/L)预处理后,tBHQ 25μmol/L组细胞活力分别恢复至(101.44±1.63)%和(92.06±9.95)%,tBHQ 50μmol/L组细胞活力分别恢复至(98.88±2.03)%和(91.12±7.87)%;NaAsO2(25、50μmol/L)单独处理后,各组细胞内ROS水平明显上升(P<0.01),分别为对照组的1.64和3.86倍;细胞经tBHQ(25、50μmol/L)预处理后,tBHQ 25μmol/L组ROS水平分别下降为对照组的0.95和1.87倍,tBHQ 50μmol/L组ROS水平分别下降为对照组的0.79和1.69倍;NaAsO2(25、50μmol/L)单独处理后,细胞内MDA含量明显上升(P<0.01),分别为2.28、2.96 nmol/(mgμprot);细胞经tBHQ(25、50μmol/L)预处理后,tBHQ 25μmol/L组MDA含量分别下降为2.05、2.43 nmol/(mgμprot),tBHQ 50μmol/L组MDA含量分别下降为2.10、2.60 nmol/(mg.prot)。结论 tBHQ对无机砷致人皮肤角质细胞损伤具有保护作用,且具有一定的剂量-反应关系。
Objective To investigate the protective effect of t-butyl hydroquinone (tBHQ) on human skin keratinocyte cell line HaCaT induced by sodium arsenite (NaAsO2). Methods The cell viability was detected by Alamar Blue reduction assay. The production of reactive oxygen species (ROS) was detected by fluorescent probe 2 ’, 7’-diacetyl dichlorofluorescein (DCFH-DA). Thiobarbituric acid TBA method was used to detect the formation of malondialdehyde (MDA), a lipid peroxide degradation product in cells. Results After treated with NaAsO2 (25 and 50μmol / L), the viability of cells in each group was significantly decreased (82% and 59%, respectively) after pretreatment with tBHQ (25 and 50μmol / L) The cell viability in L group was restored to (101.44 ± 1.63)% and (92.06 ± 9.95)%, and the cell viability in tBHQ 50μmol / L group was restored to (98.88 ± 2.03)% and (91.12 ± 7.87)%, respectively (P <0.01), which were 1.64 and 3.86 times higher than that of the control group respectively. After pretreated with tBHQ (25 and 50μmol / L), tBHQ 25μmol / L ROS levels decreased to 0.95 and 1.87 folds in the control group respectively, while the levels of ROS in the 50 μg / L tBHQ group decreased to 0.79 and 1.69 folds of those in the control group respectively. The content of MDA in the cells treated with NaAsO2 alone (25 and 50 μmol / L) The content of MDA in tBHQ 25μmol / L group decreased to 2.05,2.43 nmol / (mgμprot), tBHQ The content of MDA in 50μmol / L group decreased to 2.10 and 2.60 nmol / (mg.prot) respectively. Conclusion tBHQ has a protective effect on human skin keratinocytes induced by inorganic arsenic, and has a dose-response relationship.