MAPK信号通路调控长波紫外线诱导的皮肤成纤维细胞组织蛋白酶K表达  

MAPK pathway regulates ultraviolet A-induced cathepsin K expression in human dermal fibroblasts

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

机构地区：[1]中山大学附属第三医院皮肤科,广州510630

出　　处：《中华皮肤科杂志》2014年第8期543-547,共5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基金（2012B031800057）;2010年中国医师协会-宝洁基金;2011年中国医师协会-资生堂DQ基金

摘　　要：目的 研究MAPK信号通路调控长波紫外线（UVA）诱导皮肤成纤维细胞组织蛋白酶K（CatK）的表达.方法 原代培养的皮肤成纤维细胞取自儿童包皮.Western印迹检测10 J/cm^2 UVA照射前及照射后0.75、1.5、3和6h皮肤成纤维细胞中磷酸化JNK（p-JNK）、JNK、磷酸化P38（p-P38）、P38蛋白的表达.800 nmol/L SP600125（SP）、10 μmol/L SB203580（SB）分别孵育皮肤成纤维细胞,实验分为无UVA照射的对照组、SP组、SB组及10 J/cm^2 UVA照射的对照（UVA）组、UVA-SP组、UVA-SB组.先以Western印迹检测各组UVA照射后1.5 h磷酸化c-Jun （p-c-Jun）和磷酸化MAPKAPK2 （p-MAPKAPK2）表达,再以RT-PCR和Western印迹检测各组照射后48 h CatK mRNA、蛋白的表达.结果 皮肤成纤维细胞在UVA照射后0.75、1.5 h,p-JNK表达的灰度值分别为4.77±0.19和4.68土0.09,p-P38分别为2.44±0.13、2.30±0.04,均较照射前（p-JNK为3.2±0.27,p-P38为1.61±0.08）显著升高（均P＜0.05）;而在照射后3、6h的表达与照射前相比差异无统计学意义（P＞0.05）.p-c-Jun在UVA-SP组表达（2.55±0.48）、p-MAPKAPK2在UVA-SB组表达（1.16±0.12）均显著低于UVA组（分别为4.85±0.96和2.46±0.09）,均P＜0.05.UVA-SP组、UVA-SB组CatK mRNA、蛋白的表达分别降为UVA组的38.9％、28.7％和55.7％、49.6％（P＜0.05）,UVA-SP组CatKmRNA、蛋白的表达也均显著低于UVA-SB组（P＜0.05）.结论 JNK和P38信号通路在调控UVA上调的皮肤成纤维细胞CatK表达中起重要作用.Objective To investigate whether ultraviolet A UVA）-induced CatK expression is regulated by the mitogen-activated protein kinases （MAPK） signaling pathway in human dermal fibroblasts in vitro.Methods Human dermal fibroblasts were obtained from circumcised foreskin of children,and subjected to primary culture.After several passages of subculture,some fibroblasts were irradiated with UVA at a dose of 10 J/cm^2.Western blot was performed to measure the expressions of total and phosphorylated JNK （t-and p-JNK） and P38 （t-and p-P38） at 0.75,1.5,3 and 6 hours after the irradiation.Some fibroblasts were divided into six groups：control group receiving no treatment,SP group treated with SP600125 of 800 nmol/L,SB group treated with SB203580 of 10 μmol/L,UVA group irradiated with UVA at a dose of 10 J/cm^2,UVA-SP group treated with SP600125 for 1 hour before and for 1.5 or 48 hours after UVA irradiation at 10 J/cm^2,UVA-SB group treated with SB203580 for 1 hour before and for 1.5 or 48 hours after UVA radiation at 10 J/cm^2.Subsequently,Western blot was performed to determine the expressions of p-c-Jun and p-MAPKAPK2 in these groups at 1.5 hours after the UVA irradiation,and reverse transcription （RT）-PCR and Western blot to detect the mRNA and protein expressions of CatK at 48 hours after the UVA irradiation,respectively.Statistical analysis was carried out by t test,one way analysis of variance and least significant difference （LSD）-t test.Results The expression levels （gray values） of p-JNK and p-P38 were significantly increased at 0.75 hour （4.77 ± 0.19 and 2.44 ± 0.13 respectively,both P ＜ 0.05） and 1.5 hours （4.68 ± 0.09 and 2.30 ± 0.04 respectively,both P ＜ 0.05）,but showed no significant changes at 3 hours （both P ＞ 0.05） and 6 hours （both P ＞ 0.05） after the UVA irradiation compared with those before the irradiation （3.2 ± 0.27 and 1.61 ± 0.08 respectively）.A significant decrease was observed in the expression of p-c-Jun in the UVA-SP group and p-MAPKAP

关 键 词：成纤维细胞 组织蛋白酶类 MAP激酶信号系统 紫外线 皮肤衰老 

分 类 号：R758.1[医药卫生—皮肤病学与性病学]