UVA对成纤维细胞和HaCaT细胞形态及诱导型一氧化氮合酶产生水平的影响

Influences of UVA irradiation on the cell morphology and iNOS expression of cultured human fibroblasts and HaCaT cells

目的比较5 J/cm^2长波紫外线(UVA)照射对人皮肤成纤维细胞和HaCaT细胞形态、数目和诱导型一氧化氮合酶(iNOS)产生的影响。方法5 J/cm^2 UVA分别照射人成纤维细胞和HaCaT细胞后24 h,48 h和72 h,倒置相差显微镜下观察细胞形态和数目变化,并用逆转录聚合酶链反应(RT-PCR)和细胞免疫组化方法检测两种细胞iNOS mRNA和蛋白表达的情况。结果人成纤维细胞在照射后3个时间点细胞均出现皱缩,24 h细胞数目即明显低于照射前(P<0.05),iNOS mRNA和蛋白在照射后24 h有高峰表达;HaCaT细胞在3个时间点细胞形态未见明显改变,细胞数目在照射后48 h才明显低于照射前(P<0.05),且iNOS mRNA和蛋白在照射后48 h有高峰表达。结论5 J/cm^2 UVA照射后人成纤维细胞比HaCaT细胞更易受到损伤,iNOS高峰表达出现更早,且持续时间更长。

Objective To compare the effects of UVA irradiation on cell morphology, quantity and expression of inducible nitric oxide synthase （iNOS） mRNA and protein in human fibroblasts versus a keratinocyte cell line HaCaT. Methods Human fibroblasts and HaCaT cells were cultured and irradiated by 5 J/cm^2 UVA. Then, at 24, 48 and 72 h after the stimulation, the cell morphology was observed under an inverted microscope, and iNOS mRNA and protein were measured by RT-PCR and immunohistochemistry method, respectively. Results After the irradiation, human fibroblasts showed shrinkage at the three time points, the quantities of the cells began to decrease significantly at 24 h （P 〈 0.05 ）, and iNOS mRNA and protein expression peaked at 24 h. However, HaCaT cells seemed not to shrink at any of the three time points. Moreover, the quantity of HaCaT cells was observed to decrease significantly at 48 h after the irradiation, and iNOS mRNA and protein expression also peaked at this time point. Conclusions As compared with HaCaT cells, human fibroblasts are more vulnerable to UVA, and their iNOS expression peaks earlier and lasts longer after UVA irradiation.