热处理对UVB辐射后体外培养人黑素细胞活性的影响  

Effects of heat treatment on the activity of cultured human melanocytes irradiated by ultraviolet B in vitro

作　　者：宋文婷[1] 赵广[2] 牛建荣[2] 孟如松[2] 张悦[2] 邵丽芳[2] 成玉[2] 闫文厅[1] 

机构地区：[1]第四军医大学西京医院皮肤科,西安710032 [2]空军总医院皮肤科

出　　处：《中华皮肤科杂志》2012年第5期340-343,共4页Chinese Journal of Dermatology

摘　　要：目的探讨热处理与窄谱中波紫外线（NB—UVB）联合作用对正常人黑素细胞活性的影响。方法分别以20、30、50、70、90、120、180mJ／cm^2NB—UVB辐射体外培养的正常人黑素细胞，采用四甲基偶氮唑蓝比色法（MTT）检测细胞增殖活性并选择最佳照射剂量；以42℃，1h为热处理干预剂量，将黑素细胞分为4组：正常对照组、UVB组、加热组、UVB＋加热组，连续干预3d，以左旋多巴为底物测定酪氨酸酶活性，NaOH法测定黑素含量，流式细胞仪检测细胞周期变化。结果NB—UVB照射呈剂量依赖性减少黑素细胞存活率，选择50mJ／cm^2作为最佳照射剂量；黑素细胞经不同干预后，UVB组、UVB＋加热组的酪氨酸酶活性分别为0．244±0．018、0．310±0．015，较对照组（0．235±0．018）分别增长3．8％和31．9％（P〈0．05），两组黑素含量分别为0．201±0．016、0．286±0．019，较对照组（0．171±0．016）分别增长17．5％和67．3％（P〈0．05）；UVB组和uvB＋加热组处于G，期的黑素细胞较对照组分别减少23．94％和33．51％（P〈0．05），s期细胞分别增加15．35％（P〈0．05）和17．76％（P〉0．05），G2期分别增加11．93％（P〈0．05）和16．08％（P〉0．05）。结论热处理与NB—UVB可以协同增加黑素细胞的酪氨酸酶活性，促进黑素合成及细胞的增殖分化。Objective To investigate the effect of heat treatment combined with narrow band ultraviolet B （NB-UVB） on cultured normal human melanocytes in vitro. Methods Melanocytes were isolated from the foreskin of normal human, cultured in vitro, and irradiated with NB-UVB of different doses （20, 30, 50, 70, 90, 120 and 180 mJ/cm^2）. Then, MTT assay was performed to evaluate the proliferation and activity of melanocytes to determine the optimal dose of UVB for the next experiment. Melanocytes were classified into 3 groups to be treated with heat at 42 ℃ for 1 hour （heat group）, irradiated with UVB at 50 mJ/cm^-1 （UVB group）, or irradiated with UVB at 50 mJ/cm^2 followed by heat treatment at 42 ℃ for 1 hour （combination group）, daily for 3 successive days; those receiving no treatment served as the control. After 24-hour culture following the last treatment, tyrosinase activity was evaluated with L-dopa as the substrate, melanin content was detected by NaOH assay, and cell cycle stages were determined by flow cytometry. Results NB-UVB irradiation decreased the viability of melanocytes in a dose-dependent manner, and the optimum dose of UVB was 50 mJ/cm2. The tyrosinase activity of melanocytes was 0.244 ± 0.018 and 0.310 ± 0.015 respectively in the UVB group and combination group, and increased by 3.8% （P 〈 0.05） and 31.9% （P 〈 0.05） respectively compared with the control group （0.235 ± 0.018）; the melanin content was 0.201 ± 0.016 and 0.286± 0.019, respectively in the UVB group and combination group, and increased by 17.5% （P 〈 0.05） and 67.3% （P 〈 0.05） compared with the control group （0.171 ±0.016）. In comparison with the control group, the percentage of melanocytes in G1 phase was decreased by 23.94% in the UVB group （P 〈 0.05） and 33.51% in the combination group （P 〈 0.05）, while that in S phase and G2 phase increased by 15.35% （P 〈 0.05） and 11.93% （P 〈 0.05）, respectively in the UVB group, and 17.76% （P 〉 0.05） and 16.08% （P

关 键 词：热 紫外线 黑素细胞 细胞周期 

分 类 号：R758.41[医药卫生—皮肤病学与性病学]