摘要
目的:观察不同剂量长波紫外线(UVA)对人皮肤成纤维细胞增殖及诱导型一氧化氮合酶(iNOS)/一氧化氮(NO)系统的影响,探讨皮肤光老化机制。方法:用1,5,10 J/cm2剂量UVA照射培养的人原代皮肤成纤维细胞。分别采用四唑盐比色实验(MTT法)、逆转录-聚合酶链反应(RT-PCR)、Western印迹以及Griess反应等技术检测UVA照射后培养不同时间(24,48和72 h)人皮肤成纤维细胞增殖情况、iNOS mRNA和蛋白表达水平及NO生成量。结果:正常人成纤维细胞在72 h之内随培养时间延长细胞存活率增加,仅检测到低水平iNOS表达和NO生成,但在5和10 J/cm2剂量UVA照射后各时间点成纤维细胞存活数下降,iNOS mRNA/蛋白表达水平和NO生成量随着UVA剂量的增加升高,与同一时间点对照组和1 J/cm2UVA照射组相比,差异有统计学意义(P<0.01或P<0.05),而且以各剂量UVA照射后24 h时间点细胞存活数下降、iNOS mRNA/蛋白表达水平和NO生成量增高最为明显,与同剂量UVA照射后48 h和72 h组相比,差异有统计学意义(P<0.01)。结论:UVA抑制人皮肤成纤维细胞的增殖可能与其诱导iNOS基因的表达和NO的分泌有关。
Objective To investigate the effect of different dosages of ultraviolet A（UVA） on the proliferation and inducible nitric oxide synthase（iNOS）/nitric oxide（NO） system of human skin fibroblasts and to study the mechanism of skin photoaging.Methods Fibroblasts from normal skin cultured in vitro were irradiated with 1,5,and 10 J/cm2 of UVA,respectively.The proliferation activity,expression of iNOS mRNA/protein and NO production of human skin fibroblasts at different time points after irradiation of different dosages of UVA were measured by MTT assay,RT-PCR,Western blotting,and Griess reaction,respectively.Results The survival rate of normal fibroblasts with time increase during 72 h.Low level of iNOS mRNA/protein expression and NO production was detected in normal human skin fibroblasts.But at each time point after 5 and 10 J/cm2 dosage of UVA,the decrease of cell survival rate and the increase of iNOS mRNA/protein expression and NO production became more significant than those in the control group at the same time and 1 J/cm2 UVA-irradiated group.There were significant differences（P0.01 or P0.05）,which were most significant at 24 h after UVA irradiation.There was significant difference compared with that at 48 h and 72 h after the same UVA dosage（P0.01）.Conclusion UVA can inhibit the proliferation activity of human skin fibroblasts.It might be related to the up-regulation of iNOS gene expression and the over-secretion of NO induced by UVA.
出处
《中南大学学报（医学版）》
CAS
CSCD
北大核心
2009年第8期705-711,共7页
Journal of Central South University ：Medical Science
关键词
长波紫外线
成纤维细胞
存活率
诱导型一氧化氮合酶
一氧化氮
ultraviolet A（UVA）
fibroblasts
survival rate
inducible nitric oxide synthase（iNOS）
nitric oxide（NO）
