UVA照射对皮肤成纤维细胞组织蛋白酶K表达的影响  

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

作　　者：许庆芳[1] 侯巍[1] 刘晨[1] 郑跃[1] 龚子鉴[1] 赖维[1] 

机构地区：[1]中山大学附属第三医院皮肤性病科,广州510630

出　　处：《中华皮肤科杂志》2013年第9期652-655,共4页Chinese Journal of Dermatology

基　　金：国家自然科学基金（81171523）;广东省自然科学基金（10151008901000117）;广东省科技计划项目（20128031800057）

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10J／em2UVA照射皮肤成纤维细胞，在照射后24、48、72h提取照射组和对照组细胞蛋白和mRNA。再以10J／cm2、20J／cm2、30J／cm2UVA照射皮肤成纤维细胞，在照射后48h收集细胞。用RT—PCR和Western印迹法分别检测各组间CatKmRNA、蛋白的表达的差异。结果10J／cm2UVA照射后1、2、3d，照射组CatKmRNA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、30J／cmzUVA照射后，48h皮肤成纤维细胞中CatKmRNA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。Objective To observe the expression changes of cathepsin K （CatK） in human dermal fibroblasts at different time points after different doses of uhraviolet 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/em2 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 clay 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-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-regulated in human dermal fibroblasts after UVA irradiation.

关 键 词：成纤维细胞 弹性组织 组织蛋白酶K 紫外线 

分 类 号：R758.1[医药卫生—皮肤病学与性病学]