中华皮肤科杂志 ›› 2013, Vol. 46 ›› Issue (9): 652-655.

• 论著 • 上一篇    下一篇

UVA照射对皮肤成纤维细胞组织蛋白酶K表达的影响

许庆芳1,侯巍2,刘晨3,郑跃1,龚子鉴4,赖维5   

  1. 1. 中山大学附属第三医院皮肤科
    2. 新疆医科大学第一附属医院
    3. 深圳市人民医院
    4. 中山大学附属第三医院
    5. 广州中山大学附属第三医院皮肤科
  • 收稿日期:2012-11-07 修回日期:2013-01-15 出版日期:2013-09-15 发布日期:2013-09-01
  • 通讯作者: 赖维 E-mail:drlaiwei@163.com
  • 基金资助:
    ：Cathepsin K在光老化皮肤弹性纤维变性堆积机制中的作用;MAPK 信号通路在UVA致皮肤成纤维细胞表达组织蛋白酶K变化中的调控研究;广东省自然科学基金

Effects of ultraviolet A irradiation on the expression of cathepsin K in human dermal fibroblasts

qingfang XUWei HOU2,Chen LIU3,Zheng Yue4,Zi-Jian GONG5,Wei Lai   

  • Received:2012-11-07 Revised:2013-01-15 Online:2013-09-15 Published:2013-09-01
  • Contact: Wei Lai E-mail:drlaiwei@163.com

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摘要： 【摘要】 目的 探讨皮肤成纤维细胞中组织蛋白酶K（Cathepsin K，CatK）的表达随UVA照射后时间及其剂量的变化。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。先以10 J/cm2 UVA照射皮肤成纤维细胞，在照射后24、48、72 h提取照射组和对照组细胞蛋白和mRNA。再以10 J/cm2、20 J/cm2、30 J/cm2 UVA照射皮肤成纤维细胞，在照射后48 h收集细胞。用RT-PCR和Western印迹法分别检测各组间CatK mRNA、蛋白的表达的差异。结果 10 J/cm2 UVA照射后1、2、3 d，照射组CatK mRNA表达分别为0.351 ± 0.038（对照组0.177 ± 0.006）、0.510 ± 0.017（对照组0.176 ± 0.002）、0.313 ± 0.012（对照组0.173 ± 0.002）。照射组CatK蛋白灰度值分别为1.76 ± 0.27（对照组0.82 ± 0.45）、2.97 ± 0.36（对照组1.58 ± 0.15）、2.23 ± 0.14（对照组1.29 ± 0.32），照射组CatKmRNA、蛋白表达均较对照组升高（P < 0.05），且以照射后第2天表达最高。10、20、30 J/cm2 UVA照射后，48 h皮肤成纤维细胞中CatK mRNA表达分别为对照组的2.34、2.91、3.18倍，CatK蛋白表达分别为对照组的1.77、2.82、3.64倍，均随UVA剂量的升高而增加，其差异有统计学意义（P < 0.05）。结论 CatK在UVA照射后皮肤成纤维细胞中的表达升高。 【关键词】 成纤维细胞； 弹性组织； 组织蛋白酶K； 紫外线

关键词: 成纤维细胞, 组织蛋白酶K, 弹性组织, 紫外线

Abstract: XU Qing?鄄fang, HOU Wei, LIU Chen, ZHENG Yue, GONG Zi?鄄jian, LAI Wei. Department of Dermatology and Venereology, Third Affiliated Hospital, Sun Yat?鄄sen University, Guangzhou 510630, China Corresponding author: LAI Wei, Email: drlaiwei@163.com 【Abstract】 Objective To observe the expression changes of cathepsin K （CatK） in human dermal fibroblasts at different time points after different doses of ultraviolet A （UVA） irradiation. Methods Dermal fibroblasts were isolated from circumcised foreskins of children, and subjected to primary culture and subculture. Cells at third - tenth passage were used in the following experiment. Some fibroblasts were irradiated with UVA of 10 J/cm2 and collected at 24, 48 and 72 hours separately after the irradiation, and some fibroblasts were irradiated with UVA of 10, 20 and 30 J/cm2 separately and harvested 48 hours later. The fibroblasts receiving no irradiation served as the control group. Reverse transcription PCR and Western blot were carried out to detect the mRNA and protein expressions of CatK in fibroblasts, respectively. Results Compared with the control fibroblasts, those irradiated with UVA of 10 J/cm2 showed a significant elevation in the mRNA and protein expression levels of CatK on day 1 （0.351 ± 0.038 vs. 0.177 ± 0.006, 1.76 ± 0.27 vs. 0.82 ± 0.45, respectively, both P < 0.05）, day 2 （0.510 ± 0.017 vs. 0.176 ± 0.002, 2.97 ± 0.36 vs. 1.58 ± 0.15, respectively, both P < 0.05） and day 3 （0.313 ± 0.012 vs. 0.173 ± 0.002, 2.23 ± 0.14 vs. 1.29 ± 0.32, respectively, both P < 0.05）, with the highest expressions of CatK mRNA and protein observed on day 2. Within the range of 10 － 30 J/cm2, UVA enhanced the CatK mRNA and protein expression levels in a dose?鄄dependent manner. In detail, at 48 hours after the irradiation with UVA of 10, 20 and 30 J/cm2, the CatK mRNA expression level in the irradiated fibroblasts was 2.34, 2.91 and 3.18 times, and the CatK protein expression level 1.77, 2.82 and 3.64 times, respectively, that in the control fibroblasts （all P < 0.05）. Conclusion The expression of CatK is up?鄄regulated in human dermal fibroblasts after UVA irradiation. 【Key words】 Fibroblasts; Elastic tissue; Cathepsin K; Ultraviolet rays

Key words: Cathepsin K