组织工程与重建外科杂志 ›› 2017, Vol. 13 ›› Issue (2): 77-81.doi: 10.3969/j.issn.1673-0364.2017.02.005

• 论著 • 上一篇    下一篇

藏红花素对光老化皮肤成纤维细胞的影响

邓明武,张溢晨,李东,于子优,王祥盛,张文杰   

  1. 上海交通大学医学院附属第九人民医院整复外科;上海市组织工程重点实验室;上海星河湾双语学校
  • 发布日期:2020-07-23

Effects of Crocin on Photoaged Skin Fibroblasts

DENG Mingwu,ZHANF Yicheng,LI Dong,YU Ziyou,WANF Xiangsheng,ZHANG Wenjie   

  • Published:2020-07-23

摘要: 目的 探讨藏红花素对光老化皮肤成纤维细胞的影响,期望从细胞水平阐明藏红花素抗皮肤光老化的作用与机理,为临床应用提供依据。方法 (1)分离培养人皮肤成纤维细胞,以不同浓度藏红花素(0、50、100、200μM)处理皮肤成纤维细胞,培养72 h后,应用CCK-8试剂盒检测细胞增殖情况。(2)以不同剂量紫外线(25、50、100 m J/cm~2)照射皮肤成纤维细胞,72 h后应用CCK-8试剂盒检测细胞增殖情况。(3)以不同浓度藏红花素(0、12.5、25、50、100μM)处理皮肤成纤维细胞24 h,紫外线(100 m J/cm~2)照射后即刻,应用流式细胞仪检测细胞内活性氧簇含量;照射后48 h后,应用流式细胞仪检测细胞周期;照射后72 h后,应用CCK-8试剂盒检测细胞增殖情况,并提取细胞RNA,应用RT-PCR技术检测细胞Ⅰ型胶原的表达。结果 (1)低浓度藏红花素(50、100μM)对皮肤成纤维细胞增殖无显著影响,高浓度藏红花素(200μM)可促进皮肤成纤维细胞增殖。(2)紫外线照射可显著抑制皮肤成纤维细胞增殖,并呈量效依赖关系。(3)紫外线照射提高细胞活性氧簇产生、抑制细胞增殖、抑制Ⅰ型胶原的表达,藏红花素(12.5、25、50、100μM)能减少紫外线照射引起的细胞活性氧簇产生,恢复细胞周期和Ⅰ型胶原的合成能力。结论 藏红花素通过减少细胞活性氧簇,产生抵抗紫外线照射的作用,恢复皮肤成纤维细胞细胞增殖和胶原合成能力,起到抗光老化作用。

关键词: 皮肤光老化, 藏红花素, 抗氧化

Abstract: Objective To investigate the protective effects of crocin on photoaged skin fibroblasts, and elucidate the mechanism of crocin on anti-photoaging, which could provide new evidence for clinical application of crocin. Methods 1. Human skin fibroblasts were isolated and treated with different concentration of crocin (0, 50, 100, 100μM) for 72 hours. CCK-8 assay was performed to detect cell proliferation. 2. Fibroblasts were irradiated with different doses of ultraviolet light (25, 50, 100 mJ/cm2), cell proliferation was measured by CCK-8 kit after 72 hours. 3. Fibroblasts were treated with different concentration of crocin (0, 50, 100, 100μM) for 24 hours and irradiated by ultraviolet light (100 mJ/cm2). Intracellular reactive oxygen species (ROS) were analyzed immediately by flow cytometry. Cell cycles were analyzed 48 hours after irradiation by flow cytometry. Seventy-two hours after treatment, cell proliferation was measured by CCK-8 kit, total RNA was extracted, and the expression of type Ⅰ collagen were detected by RT-PCR. Results 1. Low concentration of crocin (50, 100 μM) had no significant effect on the proliferation of skin fibroblasts while high concentration of crocin (200μM) promoted cell proliferation. 2. Ultraviolet irradiation inhibited the proliferation of skin fibroblast in a dose dependent manner. 3. Ultraviolet irradiation enhanced cellular level of ROS, and inhibited cell proliferation and the expression of typeⅠcollagen. Crocin (12.5, 25, 50, 100μM) treatment reduced the production of ROS caused by ultraviolet irradiation, and restored the cell cycle, cell proliferation, and type I collagen expression. Conclusion Crocin prevent the photoaging of dermal fibroblasts against ultraviolet irradiation by reducing the cellular level of ROS and restoring the cell cycle and type I collagen expression.

Key words: Skin photoaging, Crocin, Antioxidation

中图分类号: