重组人表皮生长因子对紫外线A照射诱导的人成纤维细胞损伤的潜在保护作用。
Potential protective effects of rhEGF against ultraviolet A irradiation-induced damages on human fibroblasts.
作者信息
Kim Ji Min, Choo Jung Eun, Kim Ki Nam, Kim Yang Seok
机构信息
Life Science Research Institute, Daewoong Pharmaceutical Co.,Ltd., Yongin, Korea.
Department of Science in Korean Medicine, Kyng Hee University, Seoul, Korea,
出版信息
Clin Cosmet Investig Dermatol. 2018 Oct 16;11:505-513. doi: 10.2147/CCID.S170697. eCollection 2018.
BACKGROUND
Ultraviolet A (UVA) rays reach the dermal skin layer and generate oxidative stress, DNA damage, and cell inflammation, which in turn lead to photo-aging and photo-carcinogenesis. While there have been many studies about the beneficial effects of topical epidermal growth factor (EGF) treatment in the healing of wounds, the effect of EGF on UVA-induced skin irritation remains unknown. To clarify the effects of EGF on UVA-induced skin damage, it was investigated whether EGF signaling can affect intracellular reactive oxygen species (ROS) and DNA damages in UVA-irradiated human dermal fibroblasts.
MATERIALS AND METHODS
Fibroblasts cultured with or without rhEGF were UVA-irradiated at 40 mJ/cm twice per day for 5 days. After the irradiation, the intracellular ROS levels and expression of catalase and superoxide dismutase-1 (SOD-1) in the fibroblasts were ascertained. Further investigation to determine the effects of EGF on UVA-induced DNA damage, including a single cell gel electrophoresis assay and an enzyme-linked immunosorbent assay (ELISA), was carried out. Moreover, the NF-κB activity was ascertained in order to investigate the effects of EGF on UVA-irradiated fibroblasts.
RESULTS
As a result, it was revealed that recombinant human EGF (rhEGF) inhibited UVA- increased intracellular ROS in the fibroblasts and increased the expression of catalase and SOD-1. Moreover, in UVA-irradiated fibroblasts, the longest DNA-damaged tails were observed, but this phenomenon was not detected in cells cotreated with both UVA and rhEGF. Also, it was observed that DNA damage induction, including that of cyclobutene pyrimidine dimers, pyrimidine (6-4) pyrimidone photoproducts, and 8-hydroxy-2-deoxyguanosine, was caused by UVA irradiation. Similar to previous results, it was downregulated by rhEGF. Furthermore, rhEGF also inhibited NF-κB gene expression and the NF-κB p65 protein level in the nucleus induced by UVA irradiation.
CONCLUSION
These results suggest that EGF might be a useful material for preventing or improving photo-aging.
背景
紫外线A（UVA）可穿透至真皮层，产生氧化应激、DNA损伤及细胞炎症，进而导致光老化和光致癌作用。虽然已有许多关于局部应用表皮生长因子（EGF）治疗伤口愈合有益效果的研究，但EGF对UVA诱导的皮肤刺激的影响尚不清楚。为阐明EGF对UVA诱导的皮肤损伤的影响，研究了EGF信号传导是否能影响UVA照射的人真皮成纤维细胞内的活性氧（ROS）和DNA损伤。
材料与方法
将培养有或没有重组人表皮生长因子（rhEGF）的成纤维细胞每天以40 mJ/cm²的剂量进行两次UVA照射，持续5天。照射后，测定成纤维细胞内的ROS水平以及过氧化氢酶和超氧化物歧化酶-1（SOD-1）的表达。进一步进行研究以确定EGF对UVA诱导的DNA损伤的影响，包括单细胞凝胶电泳分析和酶联免疫吸附测定（ELISA）。此外，测定核因子κB（NF-κB）活性以研究EGF对UVA照射的成纤维细胞的影响。
结果
结果显示，重组人表皮生长因子（rhEGF）可抑制UVA诱导的成纤维细胞内ROS增加，并增加过氧化氢酶和SOD-1的表达。此外，在UVA照射的成纤维细胞中观察到最长的DNA损伤尾，但在用UVA和rhEGF共同处理的细胞中未检测到这种现象。还观察到UVA照射会导致DNA损伤诱导，包括环丁烷嘧啶二聚体、嘧啶（6-4）嘧啶酮光产物和8-羟基-2-脱氧鸟苷。与先前结果相似，rhEGF可使其下调。此外，rhEGF还抑制UVA照射诱导的细胞核中NF-κB基因表达和NF-κB p65蛋白水平。
结论
这些结果表明，EGF可能是预防或改善光老化的有用物质。
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