摘要
目的:研究补骨脂素对UVB诱导HaCaT细胞凋亡及相关细胞因子表达的影响。方法:体外培养人皮肤角质形成细胞(HaCaT细胞),取对数生长期的细胞,随机分为空白对照组、UVB组和补骨脂素组,并用30mJ·cm-2的中波紫外线(UVB)照射UVB组和补骨脂素+UVB组,制备光老化模型。MTT法分别测定补骨脂素对正常HaCaT细胞及光老化HaCaT细胞增殖率的影响;活性氧自由基(ROS)试剂盒测定各组细胞中ROS含量;Annexin V-FITC细胞凋亡检测试剂盒测定各组细胞凋亡率;蛋白免疫印迹(Western Blot)法测定各组细胞中p53、Bcl-2及Caspase-3蛋白的表达量。结果:与空白组比较,10-7、10-6、10-5 mol·L-1补骨脂素对HaCaT细胞增殖率无显著影响(P>0.05),可用于后续实验研究。与空白组比较,UVB组ROS含量和细胞凋亡率显著升高(P<0.01),Bcl-2蛋白表达水平显著降低(P<0.01),p53及Caspase-3蛋白表达水平显著升高(P<0.01)。与模型组比较,10-7、10-6、10-5 mol·L-1补骨脂素+UVB组ROS含量和细胞凋亡率显著降低(P<0.05,P<0.01),Bcl-2蛋白表达水平显著升高(P<0.05,P<0.01),p53及Caspase-3蛋白表达水平显著下降(P<0.05,P<0.01)。结论:补骨脂素可以通过降低细胞ROS含量和凋亡率,调控凋亡相关细胞因子p53、Bcl-2及Caspase-3的表达水平,抑制UVB诱导的细胞凋亡。
Objective:To study the effect of psoralen on ultraviolet B(UVB)induced apotosis and the expression of related cytokines of human keratinocytes(HaCaT cells).Methods:The HaCaT cells were cultured in vitro,took the logarithm of the cells,cells were randomly divided into blank controlled group,UVB group and psoralen+UVB group,the UVB group and the psoralen+UVB group were set up as the light aging model by the irradiationof ultraviolet radiation B(UVB)at irradiation dose of 30 m·cm-2.Used the method of methyl thiazolyl tetrazolium(MTT)to screen the effect of psoralen on the proliferation rate of light aging HaCaT cells,the content of reactive oxygen species(ROS)in each group was determined by reactive oxygen free radical(ROS)kit,the apoptosis of HaCaT cells were determined by the Annexin V-FITC apoptosis detection kit,Western Blot method was used for the detection of effect of psoralen on p53,B-cell lymphoma-2(Bcl-2),Caspase-3 protein expression.Result:Compared with the blank group,the concentration of psoralen at 1×10-7,1×10-6,1×10-5mol?L-1 had no significant effect on cell proliferation rate(P>0.05),these could used for the future research.Compared with the blank group,the content of ROS and the apoptosis rate of the UVB group were significantly increased(P<0.01).The expression of Bcl-2 protein was significantly decreased(P<0.01),the expression of p53 and Caspase-3 protein were significantly increased(P<0.01).Compared with the UVB group,the content of ROS and the apoptosis rate of the psoralen+UVB group was significantly decreased(P<0.05,P<0.01).The expression of Bcl-2 protein was significantly increased(P<0.05,P<0.01).The expression of p53 and Caspase-3 protein were significantly decreased(P<0.05,P<0.01).Conclusions:Psoralen can reduce the content of ROS and the apoptosis rate of HaCaT cells,regulate the expression of related apotosis Cytokines p53,Bcl-2 and Caspase-3,inhibits the apoptosis of human keratinocytes induced by UVB.
作者
傅云
廖建
王业秋
张丽宏
李建民
FU Yun;LIAO Jian;WANG Yeqiu(Sichuan Integrative Medicine Hospital,Sichuan Chengdu 610041,China;Heilongjiang University of Traditional Chinese Medicine,Heilongjiang Harbin 150040,China)
出处
《四川中医》
2019年第9期30-34,共5页
Journal of Sichuan of Traditional Chinese Medicine
基金
黑龙江省应用技术研究与开发计划项目(编号PC13S15)
