【摘要】 目的研究紫外线Al(UVAl,340 nm-400 nm)对人真皮成纤维细胞氧化性损伤、对基质金属蛋白酶(MMP)和胶原蛋白表达的影响,观察氮氧化物Tempol对这些改变的保护作用。方法体外原代培养的人真皮成纤维细胞接受UVAl照射,同时加入或不加Tempol,以生化方法检测细胞超氧化物歧化酶(SOD)活性和脂质过氧化物丙二醛(MDA)含量,以半定量逆转录-聚合酶链反应及酶连免疫吸附试验测定MMP-1,MMP-3,Ⅰ型及Ⅲ型胶原mRNA或蛋白表达水平。结果15 J/cm2 UVAl照射可显著抑制细胞SOD活性,提高MDA含量,促进MMP-1,MMP-3 mRNA表达,同时显著抑制Ⅰ型和Ⅲ型胶原蛋白表达(P<0.05)。照射同时加入Tempol,在一定浓度范围,可显著抑制UVAl引起的上述改变(P<0.05)。结论Tempol在体外对UVAl引起的氧化性损伤以及真皮细胞外基质成分代谢紊乱有保护作用,因而有可能成为防治光老化的一种成分。更多还原

【Abstract】 Objective To determine the effects of Tempol, one of the nitroxides, in the presence of ultraviolet-AI (UVA1, 340 nm -400 nm) on superoxide enzyme (SOD) activity, lipid peroxidation, and expression of collagen Ⅰ, collagen Ⅲ and matrix metalloproteinases (MMP)-1, MMP-3 in human dermal fi broblasts in vitro. Methods Fibroblasts were irradiated by a single exposure to UVA1 and at the same time incubated with or without Tempol, and detected 24 h later. SOD activity and lipid peroxidation, as shown by accumulation of malondialdehyde (MDA), were detected by biochemical assay. Expression of collagen Ⅰ, collagen Ⅲ (protein levels) and MMP-1, MMP-3 (mRNA level) was examined by ELISA and semi-quantitative RT-PCR, respectively. Results A dose of 15 J/cm2 UVA1 significantly inhibited SOD activity and collagen Ⅰ , collagen Ⅲ protein levels, increased MDA level and stimulated MMP-1, MMP-3 mRNA expression (P < 0.05). Tempol could reverse these effects caused by UVA1 to some degree or completely, and with proper concentration, the results were statistically significant (P < 0.05). Conclusions Tempol exerts photoprotective activities against UVA1 irradiation in vitro. With antioxidant ability, Tempol may normalize extracellular matrix metabolism in dermis and could be used as an anti-photoageing agent.更多还原