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目的探讨长波紫外线(UVA)对人皮肤成纤维细胞增殖及诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)表达的影响,探讨白藜芦醇对此影响的防护作用及其机制。方法5J.cm-2UVA照射原代培养的成纤维细胞后予以0.01,0.1mmol.L-1浓度的白藜芦醇干预,四甲基偶氮唑盐(MTT)法检测细胞增殖活性,半定量逆转录聚合酶链反应(RT-PCR)和蛋白质印迹(Western-blot)方法检测细胞iNOS mRNA和蛋白的表达。结果UVA辐射后成纤维细胞的增殖活性(0.345±0.020)较正常对照组(0.581±0.038)降低,iNOS mRNA/蛋白的表达水平[(0.834±0.087)/(0.804±0.021)]较正常对照组(未见表达)显著增加。UVA辐射后立即予以0.01或0.1mmol.L-1浓度的白藜芦醇处理发现成纤维细胞的增殖活性上升而iNOS mRNA/蛋白的表达下调,与单独UVA辐射组相比,差异有统计学意义(P<0.05)。结论UVA可抑制体外培养的真皮成纤维细胞的增殖,诱导iNOS的表达,而白藜芦醇对UVA辐射损伤的成纤维细胞有保护作用。
Objective To investigate the effect of UVA on the proliferation of human skin fibroblasts and the expression of inducible nitric oxide synthase (iNOS), and to explore the protective effect of resveratrol on it and its mechanism. Methods The primary cultured fibroblasts were irradiated with 5J.cm-2UVA and then treated with 0.01, 0.1mmol.L-1 resveratrol. The cell proliferation activity was detected by MTT assay. Reverse transcription polymerase chain reaction (RT-PCR) and Western-blot (Western-blot) methods were used to detect the expression of iNOS mRNA and protein. Results The proliferation activity of fibroblasts after UVA irradiation (0.345±0.020) was lower than that of the normal control group (0.581±0.038), and the expression level of iNOS mRNA/protein [(0.834±0.087)/(0.804±0.021)] was higher than that of the normal control group. (No expression) Significantly increased. Treatment with resveratrol at a concentration of 0.01 or 0.1 mmol.L-1 immediately after UVA irradiation revealed that the proliferation activity of fibroblasts increased while the expression of iNOS mRNA/protein was down-regulated. Compared with the UVA irradiation alone group, the difference was statistically significant. (P<0.05). Conclusion UVA can inhibit the proliferation of cultured dermal fibroblasts and induce the expression of iNOS, while resveratrol can protect the fibroblasts damaged by UVA radiation.