CAJ | 학술논문

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℃,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.
목적 탐토열처리여착보중파자외선(NB-UVB)연합작용대정상인흑소세포활성적영향.방법 분별이20、30、50、70、90、120、180 mJ/cm2 NB-UVB복사체외배양적정상인흑소세포,채용사갑기우담서람비색법(MTT)검측세포증식활성병선택최가조사제량；이42℃,1h위열처리간예제량,장흑소세포분위4조:정상대조조、UVB조、가열조、UVB+가열조,련속간예3d,이좌선다파위저물측정락안산매활성,NaOH법측정흑소함량,류식세포의검측세포주기변화.결과 NB-UVB조사정제량의뢰성감소흑소세포존활솔,선택50 mJ/cm2작위최가조사제량；흑소세포경불동간예후,UVB조、UVB+가열조적락안산매활성분별위0.244±0.018、0.310±0.015,교대조조(0.235±0.018)분별증장3.8％화31.9％(P＜0.05),량조흑소함량분별위0.201±0.016、0.286±0.019,교대조조(0.171±0.016)분별증장17.5％화67.3％(P＜ 0.05)；UVB조화UVB+가열조처우G1기적흑소세포교대조조분별감소23.94％화33.51％(P＜ 0.05),S기세포분별증가15.35％(P＜ 0.05)화17.76％(P＞0.05),G2기분별증가11.93％(P＜ 0.05)화16.08％(P＞ 0.05).결론 열처리여NB-UVB가이협동증가흑소세포적락안산매활성,촉진흑소합성급세포적증식분화.
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,cullured 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.