中波紫外线对人角质形成细胞iNOS的作用

Level of iNOS Synthase in Cultured Human Keratinocytes effected by UVB Irradiation

目的观察中波紫外线(UVB)照射对体外培养正常人角质形成细胞(HKC)中诱导型一氧化氮合成酶(iNOS)的作用,探讨紫外线引起皮肤细胞急性损伤的发病机制。方法体外培养HKC,用免疫荧光染色和蛋白印迹技术检测照射后不同时间细胞内iNOS的水平。结果HKC经UVB照射后,免疫荧光法检测到细胞膜胞浆侧有iNOS,蛋白印迹法检测到细胞提取总蛋白中分子量为130kDa的蛋白条带可以被抗iNOS多克隆抗体所识别,二者结果相符合。结论UVB照射可诱导体外培养的HKC在胞浆内产生iNOS,iNOS蛋白的表达可能与紫外线引起的皮肤急性炎症反应有关。

Objective To observe the level of inducible nitric oxide (iNOS) synthase in cultured normal human keratinocytes after UVB irradiation and study the pathogenesis of UV induced acute damage of the skin.Methods Normal human keratinocytes were cultured and the cells were irradiated with UVB at different dosage.We detected the level of iNOS by immunofluorescent staining and Western-blotting after UVB irradiation.Results The level of iNOS protein in UVB irradiated cultured keratinocytes was identified by immunofluorescent staining,which was more prominent at the inner side of cell membrane. Western-blot also revealed a 130 kDa protein reacted with polyclonal anti-iNOS antibodies.Conclusion UVB irradiation induced iNOS synthase which may take part in the pathogenesis of UV related acute inflammation of the skin.