c-jun氨基末端激酶在UVA诱导人角质形成细胞IL-10表达中的作用

Effects of c-Jun N-terminal Kinase on Expression of Interleukin-10 in Keratinocytes Induced by Ultraviolet A

目的探讨c-jun氨基末端激酶(JNK)是否介导UVA诱导的人角质形成细胞IL-10的表达。方法以2.4J/cm2UVA照射体外培养的2×104个/ml人角质形成细胞系(HaCaT)细胞,于照射后不同时点(0～48h)收集细胞并应用免疫荧光法检测磷酸化、非磷酸化JNK的水平。以JNK抑制剂SP600125(终浓度为10μmol/L)预处理人角质形成细胞,收集UVA照射后各时点(0～48h)的细胞及培养上清液,采用RT-PCR和双抗夹心ELISA方法检测IL-10mRNA及其蛋白表达。结果照射组角质形成细胞磷酸化JNK水平在各采样时点均明显高于对照组,差异有统计学意义(P<0.05);照射组角质形成细胞非磷酸化JNK水平于0、2、12、24、48h均高于对照组,差异有统计学意义(P<0.05)。SP600125预处理的细胞于照射后的各时点均未检测到IL-10mRNA及其蛋白的表达。结论 JNK可能介导UVA诱导的人角质形成细胞IL-10的表达。

Objective To explore the effect of c-Jun N-terminal kinase （JNK）on the expression of interleukin-10 （IL-10） in keratinocytes induced by ultraviolet A （UVA）. Methods The HaCaT cells in cultured were either sham irradiated （control） or exposured to 2.4 J/cm^2 UVA radiation. The cells were collected at 0-48 h after irradiation, and JNK levels in cells were detected with the immunofluorescence. HaCaT cells were treated with SP600125 （a JNK inhibitor） before irradiation,then cells and suspended medium were collected at each time-point after irradiation, and the expression of IL-10 mRNA and protein were determined by RT-PCR and ELISA. Results Compared with control cells,irradiated cells had increased levels of phospho-JNK throughout the entire 48 h following irradiation （P〈0.05）. The level of non-phospho-JNK was also significantly increased （P〈0.05） from 0 to 48 h,except at 4 h after irradiation. The expression of IL-10 mRNA and protein were not detected in irradiated cells treated with SP600125. Conclusion JNK may mediate the expression of IL-10 in keratinocytes induced by UVA.