CAJ | 학술논문

目的探讨姜黄素对人表皮黑素细胞活性、黑素细胞迁移及c-kit mRNA相对表达量的影响.方法不同浓度(5、10、20、30 μmol/L)姜黄素对人表皮黑素细胞活性影响实验,分为阴性对照组(添加MelM-2完全培养基+细胞)、药物对照组(MelM-2完全培养基+姜黄素)、空白对照组(仅添加MelM-2完全培养基)及实验组(MelM-2完全培养基+姜黄素+细胞),用MTS法分别检测培养24 h、48 h后细胞活性.划痕实验检测黑素细胞迁移的情况,实时荧光定量PCR检测黑素细胞c-kit mRNA相对表达量.实验分为对照组(仅添加MelM-2完全培养基+细胞)及3个浓度(5、10、20 μmol/L)实验组.结果不同浓度(5、10、20、30 μmol/L)姜黄素培养基培养黑素细胞24 h、48 h时,与对照组相比,30 μmol/L组均可明显抑制黑素细胞的增殖(P＜0.05),其他浓度差异无统计学意义(均P＞ 0.05).划痕实验结果显示,与对照组相比,在培养48 h时,5、10、20 μmol/L姜黄素均能显著抑制黑素细胞迁移(均P＜0.05).实时荧光定量PCR检测结果显示,与对照组相比,经各浓度(5、10、20 μmol/L)姜黄素作用48 h后,均可明显抑制黑素细胞c-kit mRNA相对表达量(均P＜0.05).结论低浓度的姜黄素(≤20 μmol/L)对黑素细胞无明显细胞毒性,30μmol/L的姜黄素能够促进黑素细胞的凋亡.姜黄素(≤20 μmol/L)可抑制黑素细胞的迁移,并下调人表皮黑素细胞c-kit mRNA的相对表达量.
목적 탐토강황소대인표피흑소세포활성、흑소세포천이급c-kit mRNA상대표체량적영향.방법 불동농도(5、10、20、30 μmol/L)강황소대인표피흑소세포활성영향실험,분위음성대조조(첨가MelM-2완전배양기+세포)、약물대조조(MelM-2완전배양기+강황소)、공백대조조(부첨가MelM-2완전배양기)급실험조(MelM-2완전배양기+강황소+세포),용MTS법분별검측배양24 h、48 h후세포활성.화흔실험검측흑소세포천이적정황,실시형광정량PCR검측흑소세포c-kit mRNA상대표체량.실험분위대조조(부첨가MelM-2완전배양기+세포)급3개농도(5、10、20 μmol/L)실험조.결과 불동농도(5、10、20、30 μmol/L)강황소배양기배양흑소세포24 h、48 h시,여대조조상비,30 μmol/L조균가명현억제흑소세포적증식(P＜0.05),기타농도차이무통계학의의(균P＞ 0.05).화흔실험결과현시,여대조조상비,재배양48 h시,5、10、20 μmol/L강황소균능현저억제흑소세포천이(균P＜0.05).실시형광정량PCR검측결과현시,여대조조상비,경각농도(5、10、20 μmol/L)강황소작용48 h후,균가명현억제흑소세포c-kit mRNA상대표체량(균P＜0.05).결론 저농도적강황소(≤20 μmol/L)대흑소세포무명현세포독성,30μmol/L적강황소능구촉진흑소세포적조망.강황소(≤20 μmol/L)가억제흑소세포적천이,병하조인표피흑소세포c-kit mRNA적상대표체량.
Objective To explore the effect of curcumin on the activity and migration of as well as c-kit mRNA expression in melanocytes.Methods Human epidermal melanocytes were isolated from the prepuce in adolescents and subjected to primary culture.To estimate the effect of curcumin on the proliferative activity of melanocytes, some melanocytes were randomly divided into several groups to be cultured in the MelM-2 medium with or without the presence of 5, 10, 20 or 30 μmol/L curcumin, the MelM-2 medium containing curcumin of 5-30 μmol/L served as the drug control groups, and the MelM-2 medium without curcumin served as the blank control group.After 24 and 48 hours of culture, MTS assay was performed to evaluate the proliferative activity of melanocytes.Some cultured melanocytes were randomly divided into 4 groups to be cultured in the MelM-2 medium with 0, 5, 10 and 20 μmol/L curcumin respectively for 48hours.Then, wound scratch assay was conducted to estimate the migratory activity of melanocytes, and real-time fluorescence-based quantitative PCR to quantify the mRNA expression of c-kit in melanocytes.Statistical analysis was carried out by factorial design analysis of variance (ANOVA), one-way ANOVA and least significant difference (LSD)-t test.Results The proliferative activity of melanocytes was significantly decreased at 24 and 48 hours in the 30-μmol/L curcumin group compared with the negative control group (0.783 ± 0.053 vs.1.000 ± 0.018 at 24 hours, 0.637 ± 0.015 vs.0.993 ± 0.064 at 48 hours, both P ＜ 0.05), while no significant differences were observed between the other curcumin groups and the negative control group (all P ＞ 0.05).The 48-hour treatment with curcumin could significantly inhibit the migration of melanocytes in the 5-, 10-and 20-μmol/L curcumin groups compared with the control group (all P ＜ 0.05).The mRNA expression level of c-kit was also significantly reduced at 48 hours in the 5-, 10-and 20-μmol/L curcumin groups compared with the control group (1.799 ± 0.372, 1.539 ± 0.224 and 1.026 ± 0.038 vs.3.371 ± 0.352, all P ＜0.05).Conclusion Curcumin at low concentrations (≤ 20 μmol/L) has no obvious cytotoxicity against melanocytes, but can inhibit the migration of and c-kit mRNA expression in melanocytes, while curcumin at 30 μmol/L can promote the apoptosis of melanocytes.