白藜芦醇对长波紫外线照射的人成纤维细胞的保护作用

摘要/Abstract

摘要： 【摘要】目的探讨白藜芦醇对长波紫外线（UVA）照射的人成纤维细胞的保护作用及其机制。方法 0.01、0.1、0.5、1.0 mmol/L白藜芦醇分别处理正常人成纤维细胞6、24、48、72 h后，采用MTT法检测增殖活性。10 J/cm2 UVA照射细胞后立即加入0.01、0.1 mmol/L白藜芦醇，继续培养6、24、48、72 h，MTT法检测细胞的增殖活性，ELISA法检测上清液中白介素（IL）-1α、IL-1β、IL-6蛋白水平。结果 0.5、1.0 mmol/L白藜芦醇对正常人成纤维细胞增殖活性有明显抑制作用，72 h时抑制作用最为显著，其细胞存活率分别为31.99% ± 8.29%和21.15% ± 5.76%。单独UVA照射后6 h，细胞存活率为78.01% ± 12.74%，直至照射后72 h细胞存活率（80.64% ± 36.12%）仍低于空白对照组（99.95% ± 12.23%）；照射后6 h，培养上清液中IL-1α、IL-1β、IL-6蛋白水平分别升高至（58.39 ± 0.67） ng/L、（1294.37 ± 92.51） pg/L、（197.81 ± 6.37） ng/L，直至照射后72 h，IL-1β、IL-6蛋白分泌量仍处于高水平（1236.76 ± 56.49） pg/L、（215.65 ± 3.78） ng/L。成纤维细胞经UVA照射加白藜芦醇后，与单独UVA照射组比较，0.01 mmol/L白藜芦醇组在各观察点均使细胞存活率升高，作用72 h时细胞存活率升高至91.93% ± 12.90%，并持续抑制细胞分泌的IL-1α蛋白至48 h（43.89 ± 3.60 ng/L）、IL-1β蛋白至72 h（1110.12 ± 51.91 pg/L）、IL-6蛋白至72 h（201.94 ± 4.71 ng/L）；而0.1 mmol/L白藜芦醇组在各观察点对细胞存活率均无明显影响，仅持续抑制IL-6蛋白水平至24 h（182.90 ± 6.67 ng/L）。结论 0.01 mmol/L白藜芦醇对UVA照射损伤的成纤维细胞有保护作用，其机制可能与白藜芦醇抑制UVA诱导的细胞因子IL-1α、IL-1β、IL-6分泌有关。【关键词】紫外线；成纤维细胞；细胞因子类；白藜芦醇

关键词: 成纤维细胞, 紫外线, 白藜芦醇, 细胞因子类

Abstract: CHEN Huang, LI Yuan-hong, XU Xue-gang, WU Yan*. *Department of Dermatology, No.1 Hospital of China Medical University, Shenyang 110001, China Corresponding author: WU Yan, Email: jlwuyan@126.com 【Abstract】 Objective To investigate the protective effect of resveratrol on ultraviolet A （UVA）-irradiated human fibroblasts and its mechanism. Methods Fibroblasts were isolated from normal human foreskin and subjected to primary culture and four passages of subculture. Then, some fibroblasts were incubated with various concentrations （0.01, 0.1, 0.5 and 1.0 mmol/L） of resveratrol for 6, 24, 48 and 72 hours separately, followed by methyl thiazolyl tetrazolium （MTT） assay for the evaluation of cell proliferation. Some fibroblasts were classified into four groups: blank control group remaining untreated, UVA group irradiated with UVA only, 0.01 and 0.1 mmol/L resveratrol groups receiving UVA irradiation immediately followed by treatment with resveratrol of 0.01 and 0.1 mmol/L respectively. The dose of UVA irradiation was consistently 10 J/cm2 in these groups. After additional culture for 6, 24, 48 and 72 hours, MTT assay was conducted to evaluate cell proliferation, enzyme-linked immunosorbent assay （ELISA） to measure the levels of interleukin （IL）-1α, IL-1β, and IL-6 in the culture supernatant. Results Resveratrol at 0.5 and 1.0 mmol/L significantly inhibited the proliferation of fibroblasts, with the strongest inhibitory effect observed at 72 hours when the cell survival rate was 31.99% ± 8.29% and 21.15% ± 5.76%, respectively. After irradiation with UVA of 10 J/cm2, the survival rate of fibroblasts was 78.01% ± 12.74% at 6 hours and 80.64% ± 36.12% at 72 hours, compared to 100.04% ± 10.78% and 99.95% ± 12.23% in the blank control group respectively （both P < 0.05）; the supernatant levels of IL-1α, IL-1β and IL-6 were significantly increased compared with the blank control group at 6 hours （（58.39 ± 0.67） vs. （48.51 ± 6.20） ng/L, （1294.37 ± 92.51） vs. （1023.25 ± 86.40） pg/L, （197.81 ± 6.37） vs. （160.45 ± 7.19） ng/L, all P < 0.05）, and the increase still existed at 72 hours for IL-1β （（1236.76 ± 56.49） vs. （1045.55 ± 48.14） pg/L, P < 0.05） and IL-6 （（215.65 ± 3.78） vs. （195.09 ± 1.78） ng/L, P < 0.05）. Compared with the UVA group, the 0.01 mmol/L resveratrol group showed significantly higher survival rates at all the four time points （all P < 0.05）, but lower supernatant levels of IL-1α at 6, 24 and 48 （（43.89 ± 3.60） vs. （51.77 ± 1.77） ng/L, P < 0.05） hours as well as IL-1β and IL-6 at all the four time points （all P < 0.05）, while the 0.1 mmol/L resveratrol group experienced no significant changes in cell survival rate at any of the time points, with a significant decrease only in the supernatant level of IL-6 at 6 and 24 （（182.90 ± 6.67） vs. （240.62 ± 1.42） ng/L, P < 0.05） hours. In detail, the survival rate of fibroblasts was 91.93% ± 12.90%, with the supernatant level being （1110.12 ± 51.91） pg/L for IL-1β and （201.94 ± 4.71） ng/L for IL-6 at 72 hours in the 0.01 mmol/L resveratrol group, compared to 80.64% ± 36.12%, （1236.76 ± 56.49） pg/L and （215.65 ± 3.78） ng/L respectively in the UVA group （all P < 0.05）. Conclusion Resveratrol at 0.01 mmol/L has a protective effect on UVA-irradiated fibroblasts, likely by inhibiting the secretion of IL-1α, IL-1β and IL-6. 【Key words】 Ultraviolet rays; Fibroblasts; Cytokines; Resveratrol

Key words: fibrodblast, Cytokines

陈凰李远宏徐学刚吴严. 白藜芦醇对长波紫外线照射的人成纤维细胞的保护作用[J]. 中华皮肤科杂志, 2013, 46(11): 810-814.

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