CAJ | 학술논문

目的探讨卡泊三醇对黑素细胞黑素合成的影响及其作用的可能机制.方法用噻唑蓝比色法(MTT法)、酶学方法及NaOH法,观察卡泊三醇对黑素细胞增殖及色素合成的作用;RT-PCR和蛋白印迹分别检测卡泊三醇对黑素细胞酪氨酸酶基因转录和蛋白表达水平的影响.结果 10~(-9)～10~(-5) mol/L浓度范围的卡泊三醇对体外培养的黑素细胞增殖的影响与空白对照组比较差异无统计意义(P＞0.05).同样浓度范围下,10~(-9)、10~(-8)mol/L浓度的卡泊三醇能明显增强黑素细胞的酪氨酸酶活性和促进黑素细胞的黑素含量,与空白对照组比较,酪氨酸酶活性可分别升高137%、123%(P＜0.05),黑素含量分别增加40.63%、18.75%(P＜0.05).其中10~(-9) mol/L浓度的卡泊三醇增强黑素细胞的酪氨酸酶活性及促进黑素细胞的黑素含量最明显.与高浓度组、空白对照组比较,10~(-9) mol/L浓度的卡泊三醇使酪氨酸酶蛋白表达增高也最明显.较空白对照组增高270.4%(P＜0.05).结论卡泊三醇对黑素细胞增殖无影响,可增加黑素细胞酪氨酸酶蛋白表达水平,提高酪氨酸酶活性,从而促进黑素细胞的黑素合成.
목적 탐토잡박삼순대흑소세포흑소합성적영향급기작용적가능궤제.방법 용새서람비색법(MTT법)、매학방법급NaOH법,관찰잡박삼순대흑소세포증식급색소합성적작용;RT-PCR화단백인적분별검측잡박삼순대흑소세포락안산매기인전록화단백표체수평적영향.결과 10~(-9)～10~(-5) mol/L농도범위적잡박삼순대체외배양적흑소세포증식적영향여공백대조조비교차이무통계의의(P＞0.05).동양농도범위하,10~(-9)、10~(-8)mol/L농도적잡박삼순능명현증강흑소세포적락안산매활성화촉진흑소세포적흑소함량,여공백대조조비교,락안산매활성가분별승고137%、123%(P＜0.05),흑소함량분별증가40.63%、18.75%(P＜0.05).기중10~(-9) mol/L농도적잡박삼순증강흑소세포적락안산매활성급촉진흑소세포적흑소함량최명현.여고농도조、공백대조조비교,10~(-9) mol/L농도적잡박삼순사락안산매단백표체증고야최명현.교공백대조조증고270.4%(P＜0.05).결론 잡박삼순대흑소세포증식무영향,가증가흑소세포락안산매단백표체수평,제고락안산매활성,종이촉진흑소세포적흑소합성.
Objective To investigate the effect of calcipotriol on melanin synthesis by human melanocytes and its possible action mechanism.Methods Primary melanocytes were cultured with various concentrations(10~(-5),10~(-6),10~(-7),10~(-8),10~(-9),10~(-10) mol/L)of calcipotriol for 24 or 48 hours.Subsequently,MTT assay,NaoH assay.Dopa-oxidase assay,Western blot and semiquantitative RT-PCR were used to measure the cell proliferation of,melanin synthesis by.tyrosinase activity,protein and mRNA expression levels in the melanocytes.respectively.Those untreated melanocytes served as the control.Results The calcipotriol between 10~(-9) and 10~(-5) mol/L had no significant effect on the proliferation of cultured melanocytes(P＞0.05).while that of 10~(-9) and 10~(-8) mol/L increased tyrosinase activity by 137%and 123%,and enhanced melanin synthesis by 40.63%and 18.75%,respectively,compamd with untreated melanocytes(both P＜0.05).Moreover,the tyrosinase protein level increased by 270.4%(P＜0.05)in melanocytes treated with calcipotriol at 10~(-9) mol/L for 24 hours.The strongest tyrosinase activity and melanin synthesis was observed in melanocytes treated with calcipotriol of 10~(-9) moI/L.Conclusions The proliferation of melanocytes is unaffected by calcipotriol at 10~(-9) to 10~(-5) mol/L,but it can elevate the expression of tyrosinase protein,enhance tyrosinase activity,and promote melanin synthesis in melanocytes.