中波紫外线对HaCaT细胞凋亡、细胞周期变化及p16、c-myc蛋白表达的影响及阿魏酸干预作用研究

Effects of Ferulic on Ultraviolet-B Irradiation Induced Apoptosis,Cell Cycle Arrest and Protein Expression of p16 and c-myc in Human Keratinocytes

目的观察传统中药川芎的有效成份阿魏酸(ferulic)对中波紫外线(UVB)辐射引起的人表皮角质形成细胞系HaCaT细胞凋亡、细胞周期及对p16、c-myc蛋白水平变化的影响,以探讨其光保护作用与机制。方法培养HaCaT细胞,以不同剂量UVB和200mg/mL浓度的阿魏酸处理细胞,以流式细胞仪检测0、30、60和90mJ/cm^2的UVB照射后24h细胞周期和凋亡率变化;以Western blot方法检测30mJ/cm^2UVB照射后p16、c-myc蛋白的表达水平。结果UVB照射诱导HaCaT细胞产生凋亡,凋亡率呈UVB剂量增加而升高;30mJ/cm^2剂量下细胞可出现明显S期阻滞,但高剂量时S期细胞数量相对下降;照光后p16、c-myc蛋白水平均增高。加入阿魏酸处理可抑制UVB引起的上述改变。结论阿魏酸可明显抑制UVB引起的凋亡与细胞周期阻滞以及p16、c-myc蛋白表达。

Objective To evaluate the effects of ultraviolet-B（UVB）irradiation and intervention of ferulic on apoptosis, cell cycle,and also expression levels of p16 and c-myc protein in human eternal keratinocyte-HaCaT cells.Methods Subconfluent HaCaT cells were shammed or irradiated with different dosages of UVB irradiation and treated with ferulic.The cell cycle and apoptotic rate were determined with a DNA flow cytometer at 24h after 0,30,60 and 90mJ/cm2 UVB irradiation.Western blot assay was used to measure protein expression levels of p16 and c-myc after 30mJ/cm2 UVB irradiation.Results UVB-induced apoptosis of HaCaT cells ascended with a manner of dependence on the irradiated dosages（0,30,60,90mJ/cm2）;and S-phase arrest of cell cycle under 30mJ/cm2 of UVB irradiation were observed.But with UVB dosage increasing,the cell number in S-phase descended.The levels of P16 and c-myc protein expression in the UVB group was higher that those in the control group（P0.05）.Pre-treatment of ferulic could inhibited the above changes induced by UVB irradiation.Conclusion Ferulic can inhibit apoptosis,cell cycle arrest and protein expression of pl6 and c-myc is induced by UVB,which shows the photo-protective efficiency of ferulic.