中华皮肤科杂志 ›› 2012, Vol. 45 ›› Issue (5): 340-343.

• 论著 • 上一篇    下一篇

热处理对UVB辐射后体外培养人黑素细胞活性的影响

宋文婷1,赵广2,牛建荣3,孟如松2,张悦4,邵丽芳1,成玉5,闫文厅1   

  1. 1. 解放军空军总医院
    2. 中国人民解放军空军总医院皮肤科 100142
    3. 空军总医院皮肤科
    4. 空军总医院
    5.
  • 收稿日期:2011-08-25 修回日期:2011-10-24 出版日期:2012-05-15 发布日期:2012-05-03
  • 通讯作者: 赵广 E-mail:guangfen@yahoo.com

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

  • Received:2011-08-25 Revised:2011-10-24 Online:2012-05-15 Published:2012-05-03

PDF

21

赞

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、180 mJ/cm2 NB-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 ℃，1 h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3 d，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50 mJ/cm2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G1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。

关键词: 细胞周期

Abstract:

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/cm2）. 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/cm2 （UVB group）, or irradiated with UVB at 50 mJ/cm2 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 > 0.05）, respectively in the heat group. Conclusion Heat treatment and NB-UVB can synergistically enhance the tyrosinase activity and accelerate melanogenesis, proliferation and differentiation, of melanocytes.

Key words: Cell Cycle