紫外线诱导HaCaT细胞IL10 mRNA及蛋白表达

目的探讨紫外线致机体免疫抑制的作用机制。方法体外培养人角质形成细胞系（HaCaT）,以0.6,1.2,1.8,2.4,3.0,3.6 J/cm^2长波紫外线（UVA）照射,采用四甲基偶氮噻唑蓝（MTT）法检测细胞活力。以2.4 J/cm^2UVA照射HaCaT细胞,于0,2,4,12,24,48 h收集细胞及培养掖,分别采用RT-PCR和双抗夹心ELISA方法,检测白细胞介素10（IL10）mRNA及其蛋白表达。结果3.0,3.6 J/cm^2UVA照射使体外培养的HaCaT细胞活力明显降低（P〈0.05）;0.6～2.4 J/cm^2UVA照射对细胞活力无明显影响（P〉0.05）。HaCaT细胞经2.4J/cm^2UVA照射后12h,IL19 mRNA呈弱表达;照射后24h,培养液中IL10蛋白水平为（17.45±0.65）pg/ml;照射组其他各检测时点及对照组细胞未见IL10 mRNA及蛋白表达。结论正常情况下HaCaT细胞并不表达IL10,UVA照射可诱导其表达。

Gene and protein expression of interleukin 10 in UVA - irradiated HaCaT Cells

Objective To explore the mechanism of immunosuppression induced by ultraviolet.Methods Cultured HaCaT cells were sham irradiated(control) or irradiated with 0.6,1.2,1.8,2.4,3.0,3.6 J/cm2 UVA radiation,and respectively.The cell viability was determinated by MTT method.In addition,cultured HaCaT cells were either sham irradiated(control) or exposured to 2.4 J/cm2 UVA radiation.Cells and suspended medium were collected at 0,2,4,12,24,48h post-irradiation respectively.The levels of IL 10 mRNA and protein expression were detected by RT-PCR and ELISA respectively.Results Compared with control group,cell viability decreased significantly(P<0.05) in irradiated groups with 3.0 or 3.6 J/cm2 UVA radiation.There was no significant difference(P>0.05) between control and irradiated cells with less than 2.4 J/cm2 UVA radiation.Among control and irradiated cells over a 48 period,IL10 gene was expressed only at 12h and its protein was expressed only at 24h post-irradiation.Conclusion IL 10 may not be expressed in HaCaT cells,but UVA can induce the expression of IL 10 gene and protein.