阿魏酸对过氧化氢诱导A375细胞凋亡的保护作用研究

摘要

目的 观察阿魏酸对氧化应激诱导的黑素细胞凋亡的影响.方法 采用随机数字表法将A375细胞分为五组:空白对照组,模型组,阿魏酸高、中、低浓度组.空白对照组常规培养,模型组和阿魏酸各浓度组培养基中加入过氧化氢,使其终浓度为0.1 mmol/L,处理4h.采用荧光染料DAPI染色细胞,荧光显微镜观察各组细胞凋亡情况,Caspase活性试剂盒测定各组细胞Caspase活性.结果 与空白对照组相比,模型组Caspase-3、8、9活性水平显著升高,差异有统计学意义(P＜0.01).模型组细胞核、细胞质内可见致密浓染的颗粒,松散淡染的新月体或环状荧光,提示过氧化氢可诱导A375细胞凋亡.与模型组比较,阿魏酸高浓度组可显著下调Caspase-3和Caspase-8的表达,差异有统计学意义(P＜ 0.05或P＜0.01),但对Caspase-9作用不明显.阿魏酸高剂量组细胞核呈均匀亮蓝色,无致密凝集现象,阿魏酸低、中浓度组大多数细胞细胞核出现染色质凝集和边缘化.结论 阿魏酸通过下调过氧化氢诱导的Caspase活性而起到保护黑素细胞凋亡作用.%Objective To observe the effect of ferulic acid on the apoptosis of melanocyte induced by oxidative stress.Methods According to random number table,A375 cells were divided into five groups:control group,model group,and high,middle,low concentrations of ferulic acid groups.Cells in the control group were cultured normally,while peroxide hydrogen was added into the cultured A375 cells of model group and high,middle,low concentrations of ferulic acid groups at the final concentration of 0.1 mmol/L,duration for 4 hours.The cells were stained with fluorescent dye DAPI.The apoptosis of each group was observed by fluorescence microscope.Caspase active kit was used to measure the activity of Caspase in each group.Results Compared with the control group,the activities of Caspase-3,8 and 9 in the model group were significantly increased,with statistically significant differences (P ＜ 0.01).In the model group,dense and concentrated particles and the loose light colored crescent or annular fluorescence could be seen in the nucleus and cytoplasm,which implied that hydrogen peroxide can induce apoptosis of A375 cells.Compared with the model group,the expression of Caspase-3 and Caspase-8 were significantly reduced in the high does concentration of ferulic acid group,with statistically significant differences (P ＜ 0.05 or P ＜ 0.01),but the effect on Caspase-9 was not obvious.The nucleus of cells in the high does concentration of ferulic acid group were even bright blue,without the phenomenon of dense condensation.There were chromatin agglutination and marginalization of most cell nucleus in the low and medium does concentrations offerulic acid groups.Conclusion Ferulic acid can protect the apoptosis of melanocyte through lower the activity of Caspase induced by hydrogen peroxide.