长波紫外线对人皮肤成纤维细胞自噬水平的影响  

Effects of ultraviolet A on autophagy in human skin fibroblasts

作　　者：郑云鹏[1] 陈旭[1] 黄丹[1] 徐松[1] 顾恒[1] 

机构地区：[1]中国医学科学院北京协和医学院皮肤病研究所理疗科,南京210042

出　　处：《中华皮肤科杂志》2015年第11期782-786,共5页Chinese Journal of Dermatology

基　　金：国家自然科学基金（81371755、81171513）;江苏省自然科学基金（BK20131064）;高等学校博士学科点专项科研基金（20111106110041、20131106120046）;2014年度北京协和医学院协和青年科研基金（3332014008）;北京协和医学院研究生创新基金（2013-1002-19）

摘　　要：目的 探讨长波紫外线（UVA）对人皮肤成纤维细胞（HSF）自噬水平的影响。 方法 HSF慢性UVA照射分组：未照射组、5 J/cm^2组、10 J/cm^2组和20 J/cm^2组，每天照射1次连续4 d；HSF急性照射分组：未照射组、5 J/cm^2组、10 J/cm^2组、30 J/cm^2组和60 J/cm^2组，单次照射。采用3-（4，5-二甲基噻唑-2）-2，5-二苯基四氮唑溴盐分析法（MTT）检测细胞增殖活力。单丹酰戊二胺染色法（MDC）检测细胞自噬体。Western印迹法检测细胞LC3蛋白LC3-I向LC3-II转化分析（LC3-II/LC3-I）。 结果 与未照射组HSF比较，5、10、20 J/cm^2慢性UVA照射组细胞增殖活力降低（F = 155.5，P 〈 0.05）。与未照射组HSF比较，5、10、30、60 J/cm^2急性UVA照射后1、6和12 h，细胞增殖活力均降低（F值分别为1 335、1 649、2 774、均P 〈 0.05）。MDC法标记细胞自噬囊泡，与未照射组比较，5、10、20 J/cm^2慢性照射均可上调细胞自噬水平（F = 748.62，P 〈 0.05），而5、10、30、60 J/cm^2急性照射后1、6和12 h对细胞自噬水平无明显影响（F值分别为0.014、0.004、0.002，均P ＞ 0.05）。5、10和20 J/cm^2慢性照射组LC3-I向LC3-II转化（LC3-II/LC3-I）均增加（t 值分别为9.002、21.772、18.33，均P 〈 0.05），细胞自噬水平上调。与未照射组比较，5、10、30、60 J/cm^2急性照射后1、6和12 h，细胞LC3-I向LC3-II转化均无明显变化（F值分别为0.13、0.27、0.06，均P ＞ 0.05），对细胞自噬水平无明显影响。 结论 慢性UVA照射可上调HSF自噬水平，急性UVA照射HSF自噬无明显变化。Objective To evaluate the effects of ultraviolet A （UVA） on autophagy in human skin fibroblasts （HSFs）. Methods Cultured HSFs were randomly divided into chronic and acute UVA radiation groups. HSFs in the chronic UVA radiation groups were irradiated with UVA at 5, 10 and 20 J/cm^2 separately once a day for 4 consecutive days, with HSFs receiving no radiation serving as the chronic radiation control group; HSFs in the acute UVA radiation groups received a single session of radiation with 5, 10, 30 and 60 J/cm^2 UVA separately, with HSFs receiving no radiation serving as the acute radiation control group. After additional culture for different durations, methyl thiazolyl tetrazolium （MTT） assay was performed to evaluate the proliferative activity of HSFs, monodansylcadaverin （MDC） staining to determine autophagy levels, and Western blot analysis to track the conversion of the microtubule-associated protein 1 light chain-3 （LC3） -Ⅰto LC3-Ⅱ. Statistical analysis was carried out by using one-way analysis of variance followed by Students-Newman-Keuls （SNK） test for multiple-group comparisons and by the independent sample t test for two-group comparisons. Results The cellular proliferative activity significantly decreased in the 3 chronic radiation groups at 1 hour after the final UVA radiation compared with the chronic radiation control group （F = 155.5, P 〈 0.05）, and in the 4 acute radiation groups at 1, 6 and 12 hours after UVA radiation compared with the acute radiation control group（F = 1 335, 1 649, 2 774, all P 〈 0.05）. MDC staining showed that the autophagy levels in HSFs significantly increased in the 3 chronic radiation groups after UVA radiation compared with the chronic radiation control group （F = 748.62, P 〉 0.05）, but showed no significant changes in any of the acute radiation groups at 1, 6 or 12 hours after UVA radiation compared with the acute radiation control group （F = 0.014, 0.004, 0.002, all P 〉 0.05）. The ratio of LC3-Ⅱ to LC3-Ⅰwas signific

关 键 词：紫外线 自噬 成纤维细胞 细胞增殖 

分 类 号：R758.1[医药卫生—皮肤病学与性病学]