基础医学与临床 ›› 2017, Vol. 37 ›› Issue (12): 1741-1745.

• 研究论文 • 上一篇    下一篇

二甲双胍抑制UVA诱导的人皮肤成纤维细胞的光老化效应

林意1,周晓丽2,陈海云2,张春丽2   

  1. 1. 杭州外国语学校
    2. 南京医科大学附属友谊整形外科医院
  • 收稿日期:2016-06-21 修回日期:2017-01-16 出版日期:2017-12-05 发布日期:2017-11-29
  • 通讯作者: 张春丽 E-mail:122758476@qq.com

Repression effect of metformin on photo-damage of human skin fibroblasts induced by UVA

  • Received:2016-06-21 Revised:2017-01-16 Online:2017-12-05 Published:2017-11-29

摘要: 目的 研究二甲双胍(MF)对长波紫外线(UVA)诱导的人皮肤成纤维细胞(HSF)光老化效应的抑制作用及相关机制。方法 体外培养人皮肤成纤维细胞,分为对照组(control)、UVA组、UVA+MF组。以CCK8法检测细胞增殖;对各组细胞进行细胞衰老β-半乳糖苷酶染色;以荧光探针DCF-DA染色流式细胞仪检测细胞内ROS水平;Real-time PCR检测衰老相关基因MMP1、MMP3的mRNA相对表达量;Westernblot检测蛋白MMP1、MMP3、SOD1和SOD2的表达。结果 0.01mmol/L二甲双胍对HSF增殖无显著影响,0.1和1mmol/L二甲双胍则显著抑制HSF的细胞增殖(P<0.05)。与control组相比,UVA组β-半乳糖苷酶染色阳性率显著增高(P<0.01);ROS水平显著升高(P<0.05);MMP1和MMP3 mRNA相对表达量显著增加(P<0.01); MMP1和MMP3蛋白表达量显著增多(P<0.01);SOD1、SOD2蛋白表达显著减少(P<0.01)。与UVA组相比,UVA+MF组β-半乳糖苷酶染色阳性率显著降低(P<0.05);ROS水平显著下降(P<0.05);MMP1和MMP3的mRNA相对表达量显著降低(P<0.05);蛋白MMP1和MMP3表达量显著降低(P<0.05);蛋白SOD1表达显著增加(P<0.01);SOD2表达量增加。结论 二甲双胍可通过抑制细胞内ROS水平和相关基质金属蛋白酶的表达,提高细胞抗氧化能力,从而抑制UVA诱导的皮肤成纤维细胞的光老化效应。

关键词: 二甲双胍, UVA, 人皮肤成纤维细胞, 光老化

Abstract: Objective To determine the repression effect and mechanism of metformin on photo-damage of human skin fibroblasts induced by UVA. Methods Human skin fibroblasts were randomly divided into Control group, UVA group and UVA+MF group. The proliferation of HSF was detected by CCK-8 assay kit. SA-β-gal staining was performed to evaluate the senescence state. The level of ROS was examined by fluorescence probe DCF-DA staining using flow cytometry. Real-time PCR was used to determine mRNA expression of senescence-associated signals of MMP1 and MMP3. The protein expression of MMP1, MMP3, SOD1 and SOD2 were measured by Western blot. Results To the proliferation of HSF, 0.01mmol/L Metformin had no significant effect, but 0.1 and 1mmol/L Metformin depressed significantly(P<0.05). Compared with the Control group, it showed that UVA irradiation increased the positive rate of SA-β-gal staining(P<0.01), the level of ROS(P<0.05), mRNA and protein expression of MMP1 and MMP3 significantly(P<0.01); Also decreased the expression of SOD1 and SOD2(P<0.01). Compared with the UVA group, it showed that metformin decreased the positive rate of SA-β-gal staining(P<0.05), the level of ROS(P<0.05), mRNA and protein expression of MMP1 and MMP3 significantly(P<0.05); Also increased the expression of SOD1(P<0.01)and SOD2. Conclusion: Metfomin can repress the photo-damage of human skin fibroblasts induced by UVA via the inhibiton of ROS and metal matrix protease generation, also the improvement of cellular antioxidant capacity.

Key words: Metformin, UVA, human skin fibroblasts, photo-damage