蓝光影响人皮肤角质形成细胞、成纤维细胞及黑素细胞生物活性的初步研究  

Effect of blue light on the biological activity of human skin keratinocytes,fibroblasts and melanocytes:a preliminary study

作　　者：杨雅琪 蒋鑫 常锦绣 涂颖[1] 马彦云[2] 何黎[1] 顾华[1] Yang Yaqi;Jiang Xin;Chang Jinxiu;Tu Ying;Ma Yanyun;He Li;Gu Hua(Department of Dermatology,the First Affiliated Hospital of Kunming Medical University,Kunming 650000,China;FuDan University,Shanghai 200433,China)

机构地区：[1]昆明医科大学第一附属医院皮肤科,昆明650000 [2]复旦大学,上海200433

出　　处：《中华皮肤科杂志》2023年第12期1115-1122,共8页Chinese Journal of Dermatology

基　　金：云南省卫生和计划生育委员会医学学科带头人培养计划项目(D-2017038);云南省科技计划项目[2019FE001(-047)]。

摘　　要：目的初步探讨蓝光对人皮肤角质形成细胞、成纤维细胞与黑素细胞生物活性的影响。方法收集2021年6月至2021年12月昆明医科大学第一附属医院10例3~12岁健康儿童包皮环切术后的废弃包皮,分离表真皮后采用选择培养基分离角质形成细胞、成纤维细胞、黑素细胞。根据预实验结果采用0、5、10、20、30、40 J/cm^(2)剂量440~450 nm蓝光分别照射上述3种细胞,继续培养0、6、24、48 h,在各时间点采用CCK8法检测细胞增殖活力;酶联免疫吸附试验(ELISA)检测角质形成细胞分泌白细胞介素18(IL-18)、IL-33、神经生长因子(NGF)、粒细胞-巨噬细胞集落刺激因子(GM-CSF)和成纤维细胞分泌IL-33、角质形成细胞生长因子(KGF)的浓度;NaOH裂解法检测黑素细胞黑素合成率;Western印迹检测黑素细胞酪氨酸酶(TYR)、酪氨酸相关蛋白酶1(TRP-1)及多巴色素异构酶(DCT)的相对表达。采用双因素方差分析进行组效应、时间效应和交互效应的分析。结果各剂量蓝光照射后不同时间,角质形成细胞组间(F时间=516.20、F剂量=421.20、F交互=25.05,均P<0.003)、成纤维细胞组间(F时间=129.30、F剂量=477.80、F交互=10.91,均P<0.003)、黑素细胞组间(F时间=77.61、F剂量=138.70、F交互=3.50,均P<0.003)增殖活力差异均有统计学意义;照射后即刻,20~40 J/cm^(2)组角质形成细胞活力、成纤维细胞活力均低于0 J/cm^(2)剂量组(均P<0.003),5 J/cm^(2)组黑素细胞活力高于0 J/cm^(2)剂量组(P<0.003);细胞活力随蓝光照射剂量及继续培养时间的增加有降低趋势。ELISA检测显示,角质形成细胞分泌的IL-18、IL-33、NGF、GM-CSF及成纤维细胞分泌的IL-33、KGF浓度随蓝光照射剂量及培养时间的增加有增高趋势。各剂量蓝光照射后不同时间黑素细胞黑素合成率组间差异有统计学意义(F时间=833.50、F剂量=249.40、F交互=81.38,均P<0.003),照射后0~24 h黑素合成率随蓝光照射剂量及时间的增加�Objective To preliminarily investigate the effect of blue light on the biological activity of human skin keratinocytes,fibroblasts and melanocytes.Methods Discarded foreskin tissues were collected from 10 healthy children aged from 3 to 12 years after circumcision surgery in the First Affiliated Hospital of Kunming Medical University from June 2021 to December 2021.After epidermis-dermis separation,selective culture was performed to isolate keratinocytes,fibroblasts,and melanocytes.According to the pre-experiment results,the above three types of cells were irradiated with 440-450 nm blue light at doses of 0,5,10,20,30,and 40 J/cm^(2),and then continued to be cultured for 0,6,24,and 48 hours.Cell counting kit 8(CCK8)assay was performed to evaluate cellular proliferative activity at each time point,enzyme-linked immunosorbent assay(ELISA)to detect levels of interleukin(IL)-18,IL-33,nerve growth factor(NGF),and granulocyte-macrophage colony-stimulating factor(GM-CSF)secreted by keratinocytes,as well as levels of IL-33 and keratinocyte growth factor(KGF)secreted by fibroblasts,NaOH lysis method to determine melanin synthesis rates in melanocytes,and Western blot analysis to determine the relative expression of tyrosinase(TYR),tyrosine-related protease 1(TRP-1)and dopachrome isomerase(DCT)in melanocytes.Two-way analysis of variance was used to analyze group effects,time effects and interaction effects.Results After irradiation with blue light,the cellular proliferative activity significantly differed among different doses of blue light irradiation groups and different time points in keratinocytes(Ftime=516.20,Fdose=421.20,Finteraction=25.05,all P<0.003),fibroblasts(Ftime=129.30,Fdose=477.80,Finteraction=10.91,all P<0.003),and melanocytes(Ftime=77.61,Fdose=138.70,Finteraction=3.50,all P<0.003);immediately after irradiation,the proliferative activity of keratinocytes and fibroblasts was significantly lower in the 20-40 J/cm^(2) blue light group than in the 0 J/cm^(2) blue light group(all P<0.003),and the proliferative a

关 键 词：黑素细胞 角蛋白细胞 成纤维细胞 细胞增殖 蓝光 黑素合成 

分 类 号：R751[医药卫生—皮肤病学与性病学]