CAJ | 학술논문

目的探讨红外线(infrared,JR)照射促进体外培养的正常人表皮黑素细胞的酪氨酸酶活性及黑素含量的作用,寻找IR照射的最佳剂量.方法将体外培养的正常人表皮黑素细胞分别给予不同的红外线剂量:0 J/cm2(对照组)、20 J/cm2、60 J/cm2、80 J/cm2、100 J/cm2、140 J/cm2、240 J/cm2、320 J/cm2,采用四甲基偶氮唑蓝(MTT)法选择最佳剂量并以此作为照射条件,连续3d,检测黑素细胞的酪氨酸酶活性、酪氨酸酶活性增长率、黑素含量、红外线照射对黑素细胞影响率以及细胞周期.结果红外线照射的最佳剂量为80 J/cm2,此照射下的酪氨酸酶活性0.3601±0.0301,高于对照组0.3114±0.0341 (P＜ 0.05),酪氨酸酶活性增长率为15.64％.照射组的黑素细胞含量为0.2748±0.0243,高于对照组0.2325±0.0254(P＜0.05),红外线照射对黑素细胞黑素含量影响率18.19％.细胞周期检测结果显示,G1细胞比例明显低于对照组(P＜0.01),G2期和S期细胞比例较对照组增加(P＜0.05).结论 80 J/cm2是IR照射的理想剂量,可以增加黑素细胞的黑素含量,提高酪氨酸酶活性,促进黑素细胞分化增殖.
목적 탐토홍외선(infrared,JR)조사촉진체외배양적정상인표피흑소세포적락안산매활성급흑소함량적작용,심조IR조사적최가제량.방법 장체외배양적정상인표피흑소세포분별급여불동적홍외선제량:0 J/cm2(대조조)、20 J/cm2、60 J/cm2、80 J/cm2、100 J/cm2、140 J/cm2、240 J/cm2、320 J/cm2,채용사갑기우담서람(MTT)법선택최가제량병이차작위조사조건,련속3d,검측흑소세포적락안산매활성、락안산매활성증장솔、흑소함량、홍외선조사대흑소세포영향솔이급세포주기.결과 홍외선조사적최가제량위80 J/cm2,차조사하적락안산매활성0.3601±0.0301,고우대조조0.3114±0.0341 (P＜ 0.05),락안산매활성증장솔위15.64％.조사조적흑소세포함량위0.2748±0.0243,고우대조조0.2325±0.0254(P＜0.05),홍외선조사대흑소세포흑소함량영향솔18.19％.세포주기검측결과현시,G1세포비례명현저우대조조(P＜0.01),G2기화S기세포비례교대조조증가(P＜0.05).결론 80 J/cm2시IR조사적이상제량,가이증가흑소세포적흑소함량,제고락안산매활성,촉진흑소세포분화증식.
Objective To observe the increasing effect of infrared irradiation on tyrosinase activity and melanin content in cultured normal human epidermal melanocytes in vitro and to explore the optimal dose of infrared irradiation.Methods Epidermal melanocytes were isolated from normal human foreskin tissue,and subjected to primary culture.Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the effect of different doses(0,20,60,80,100,140,240,320 J/cm2)of infrared light on the proliferation of melanocytes and to select the optimal irradiation dose.Then,melanocytes were irradiated with infrared light at the optimal dose for 3 consecutive days followed by the determination of tyrosinase activity,melanin content,and cell cycle via dopa oxidation assay,NaOH solubilization method and flow cytometry,respectively.Results The best intervention dose of infrared light was 80 J/cm2.The tyrosinase activity (A492 nm) and melanin content (A492 nm)were 0.3601 ± 0.0301 and 0.2748 ± 0.0243 respectively in melanocytes after irradiation with infrared light of 80 J/cm2 for 3 days,significantly higher than those in unirradiated melanocytes (0.3114 ± 0.0341,0.2325 ±0.0254,respectively,both P ＜ 0.05),with an increase rate of 15.64％ and 18.19％ respectively.Cell cycle analysis revealed a decline in cell percentage in G1 phase (P ＜ 0.01 ) but a concomitant increase in cell percentage in G2 and S phase (both P ＜ 0.05) in irradiated melanocytes compared with unirradiated melanocytes.Conclusions The optimal dose of infrared light is 80 J/cm2 for the irradiation of melanocytes,and this dose of infrared light can increase melanin content,tyrosinase activity,differentiation and proliferation of melanocytes.