CAJ | 학술논문

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、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、30 J/cm2UVA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.
목적 탐토피부성섬유세포중조직단백매K(Cathepsin K,CatK)적표체수UVA조사후시간급기제량적변화.방법 원대배양적피부성섬유세포래자인동포피,10대이내적세포행후속실험.선이10 J/cm2 UVA조사피부성섬유세포,재조사후24、48、72 h제취조사조화대조조세포단백화mRNA.재이10 J/cm2、20 J/cm2、30 J/cm2 UVA조사피부성섬유세포,재조사후48 h수집세포.용RT-PCR화Western인적법분별검측각조간CatK mRNA、단백적표체적차이.결과 10 J/cm2 UVA조사후1、2、3d,조사조CatKmRNA표체분별위0.351±0.038(대조조0.177±0.006)、0.510±0.017(대조조0.176±0.002)、0.313±0.012(대조조0.173±0.002).조사조CatK단백회도치분별위1.76±0.27(대조조0.82±0.45)、2.97±0.36(대조조1.58±0.15)、2.23±0.14(대조조1.29±0.32),조사조CatKmRNA、단백표체균교대조조승고(P＜0.05),차이조사후제2천표체최고.10、20、30 J/cm2UVA조사후,48 h피부성섬유세포중CatK mRNA표체분별위대조조적2.34、2.91、3.18배,CatK단백표체분별위대조조적1.77、2.82、3.64배,균수UVA제량적승고이증가,기차이유통계학의의(P＜0.05).결론 CatK재UVA조사후피부성섬유세포중적표체승고.
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-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.