紫外线诱导HaCaT细胞IL10 mRNA及蛋白表达

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

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。

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/cm^2 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/cm^2 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/cm^2 UVA radiation. There was no significant difference（ P 〉 0.05） between control and irradiated cells with less than 2.4 J/cm^2 UVA radiation. Among control and irradiated cells over a 48 period, ILl0 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.