中华皮肤科杂志 ›› 2014, Vol. 47 ›› Issue (12): 873-876.

• 论著 • 上一篇    下一篇

长波紫外线对皮肤成纤维细胞表达和分泌组织蛋白酶G的影响

许庆芳1,侯巍2,赖维3,郑跃1,刘晨4,陆春5   

  1. 1. 中山大学附属第三医院皮肤科
    2. 乌鲁木齐市新疆医科大学第一附属医院皮肤科
    3. 广州中山大学附属第三医院皮肤科
    4. 深圳市人民医院
    5. 广州中山大学第三医院皮肤科
  • 收稿日期:2014-03-18 修回日期:2014-05-28 出版日期:2014-12-15 发布日期:2019-06-14
  • 通讯作者: 赖维 E-mail:drlaiwei@163.com
  • 基金资助:
    国家自然科学基金;广东省科技计划项目;广东省自然科学基金;CAD-资生堂DQ基金研究项目：CathepsinD对皮肤屏障的修复作用及机制研究

Effect of ultraviolet A radiation on the expression and secretion of cathepsin G by human dermal fibroblasts

  • Received:2014-03-18 Revised:2014-05-28 Online:2014-12-15 Published:2019-06-14
  • Contact: Wei Lai E-mail:drlaiwei@163.com
  • Supported by:
    ;Science and Technology Planning Project of Guangdong Province of China

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摘要： 目的 研究长波紫外线（UVA）照射对皮肤成纤维细胞组织蛋白酶G（CatG）表达和分泌的影响。 方法 原代培养的皮肤成纤维细胞来自儿童包皮，10代以内的细胞行后续实验。①以10 J/cm2 UVA照射皮肤成纤维细胞，24、48、72 h后提取照射组和对照组细胞蛋白和mRNA，并收集细胞上清液；②分别以10、20、30 J/cm2 UVA照射皮肤成纤维细胞，24 h后收集细胞和上清液。用RT-PCR 和Western印迹分别检测各组细胞CatG mRNA及蛋白的表达，ELISA检测细胞上清液CatG的含量。 结果 10 J/cm2 UVA照射后24、48、72 h，照射组细胞CatG mRNA表达分别为0.376 ± 0.014、0.308 ± 0.022和0.296 ± 0.032，对照组分别为0.183 ± 0.003、0.185 ± 0.005、0.182 ± 0.004；照射组细胞CatG蛋白灰度值分别为1.80 ± 0.12、1.41 ± 0.17和1.27 ± 0.09，对照组分别为0.96 ± 0.06、0.95 ± 0.22、1.00 ± 0.14；照射组细胞CatG mRNA、蛋白表达及细胞上清液CatG含量较相应的对照组升高（均P < 0.05），且以照射后24 h表达最高。10、20、30 J/cm2 UVA照射后24 h，皮肤成纤维细胞CatG mRNA表达分别为对照组的1.90、2.51、3.04倍，细胞CatG蛋白表达分别为对照组的1.88、3.97、4.72倍，细胞上清液CatG含量分别为对照组的1.36、1.50、1.66倍，均随UVA剂量的升高而增加，其差异有统计学意义（均P < 0.01）。 结论 急性UVA照射促进皮肤成纤维细胞表达和分泌CatG。

关键词: 成纤维细胞, 组织蛋白酶类, 紫外线, 细胞衰老

Abstract: Xu Qingfang*, Hou Wei, Lai Wei, Zheng Yue, Liu Chen, Lu Chun. *Department of Dermatology, Third Affiliated Hospital, Sun Yat-sen University, Guangzhou 510630, China Corresponding author: Lai Wei, Email: drlaiwei@163.com 【Abstract】 Objective To investigate the effect of ultraviolet A（UVA） radiation on the expression and secretion of cathepsin G （CatG） by human dermal fibroblasts. Methods Dermal fibroblasts were isolated from the foreskins of boys, and subjected to primary culture and subculture. After 10 or less passages, the fibroblasts were collected and divided into several groups to be irradiated with 10 J/cm2 UVA followed by 24, 48 and 72 hours of additional culture, or be irradiated with 10, 20 and 30 J/cm2 UVA followed by 24 hours of additional culture, with those receiving no treatment serving as the control group. Subsequently, cells and culture supernatant were collected, real time PCR and Western blot were performed to detect the expressions of CatG mRNA and protein respectively in these cells, and enzyme-linked immunosorbent assay （ELISA） was conducted to measure the expression of CatG protein in the culture supernatant of these cells. Results Compared with the control group, the fibroblasts irradiated with 10 J/cm2 UVA showed a significant increase at 24, 48 and 72 hours in the expressions of CatG mRNA （0.376 ± 0.014 vs. 0.183 ± 0.003, 0.308 ± 0.022 vs. 0.185 ± 0.005, 0.296 ± 0.032 vs. 0.182 ± 0.004, respectively, all P < 0.05） and protein （1.80 ± 0.12 vs. 0.96 ± 0.06, 1.41 ± 0.17 vs. 0.95 ± 0.22, 1.27 ± 0.09 vs. 1.00 ± 0.14, respectively, all P < 0.05）, as well as in the supernatant level of CatG protein （（161.35 ± 7.55） vs. （122.45 ± 6.46） ng/L, （141.76 ± 2.95） vs. （124.17 ± 6.15） ng/L, （139.63 ± 3.04） vs. （121.72 ± 3.17） ng/L, respectively, all P < 0.05）, with the strongest increase observed at 24 hours. At 24 hours after 10, 20 and 30 J/cm2 of UVA radiation, the expression of CatG mRNA in irradiated fibroblasts was 1.90, 2.51 and 3.04 times respectively （all P < 0.05）, the expression of CatG protein was 1.88, 3.97 and 4.72 times respectively （P < 0.05）, and the supernatant level of CatG protein was 1.36, 1.50 and 1.66 times respectively （P < 0.05）, that in the control group, and there was an increasing trend in all the above three parameters with increasing dose of UVA. Conclusion Acute UVA radiation can promote the expression and secretion of CatG by human dermal fibroblasts.

Key words: Fibroblasts, Cathepsins, Ultraviolet rays, Cell aging