tBHQ对无机砷致人皮肤角质细胞损伤保护作用

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。

Protective effects of tert-butylhydroquinone on NaAsO2-induced cytotoxicity in HaCaT cells

Objective To study protective effect of tert-butylhydroquinone in HaCaT cells on the damage induced by NaAsO2 in vitro.Methods Alamar Blue reduction rate was used to evaluate cell vitality.The level of reactive oxygen species(ROS) was detected by staining cells with 2′,7′-dichlorofluorescin diacetate(DCFH-DA).The melondiadehyde(MDA) level was measured by thiobarbituric acid(TBA) method.Results The viability of HaCaT cells was decreased by 82% and 59% after exposed to NaAsO2 at the concentrations of 25 μmol/L and 50 μmol/L but the viability was 92% and 91% for the two exposure groups pretreated wtih tert-butylhydorquinone(tBHQ).Comparted to the control groups,the level of ROS increased by 1.64 and 3.86 times in the HaCaT cells exposed to 25 μmol/L and 50 μmol/L NaAsO2 but only by 0.95 and 1.87 times for the same exposure groups pretreated with 25 μmol/L tBHQ and by 0.79 and 1.69 times for the two groups pretreated with 50 μmol/L tBHQ.The MDA levels increased significantly in HaCaT cells exposed to NaAsO2(2.28 mol/L/mg·prot for 25 μmol/L NaAsO2 exposure,2.96 mol/L/mg·prot for 50 μmol/L NaAsO2 exposure;P<0.01) and the increased levels of MDA were 2.05 μmol/L/mg·prot and 2.43 mol/L/mg·prot for 25 μmol/L pretreatment of tBHQ and 2.10 nmol/mg·prot and 260 nmol/mg·prot for 50 μmol/L/L tBHQ pretreatment groups.Conclusion tBHQ could protect HaCaT cells from damage induced by NaAsO2 in a dose-reaction manner.