小分子化合物I942在UVB照射诱导人黑素细胞树突形成及黑色素合成的促进作用

The Small Molecule Compound I942 Promotes the Formation of Dendrites and Melanin Synthesis in Human Melanocytes Induced by UVB Irradiation

目的:探究小分子化合物I942在UVB照射诱导人黑素细胞树突形成及黑色素合成的促进作用。方法:使用浓度为0、3.125、6.25、12.5、25、50μmol/L的小分子化合物I942处理正常永生化人表皮黑素细胞系PIG1,采用CCK-8法检测各组24、48、72 h时的细胞活力。将PIG1细胞分为对照组、UVB组、UVB+2.5μmol/L I942组、UVB+5μmol/L I942组、UVB+10μmol/LI942组,按照分组名称进行相应处理。多巴染色观察各组细胞形态,免疫荧光染色观察黑素小体及骨架蛋白F-actin分布情况,氢氧化钠裂解法测定黑色素含量,多巴氧化反应法测定酪氨酸酶活性。qRT-PCR、WB检测树突形态相关、黑色素合成相关mRNA与蛋白表达水平。结果:浓度为0、3.125、6.25、12.5、25、50μmol/L的小分子化合物I942对PIG1细胞活力无明显影响(P>0.05)。与对照组相比,UVB组、UVB+2.5μmol/L I942组、UVB+5μmol/L I942组、UVB+10μmol/L I942组细胞树突数量、gp100、F-actin、黑色素含量、酪氨酸酶活性、MITF、TYR、TRP-1、TRP-2、Rac1mRNA与蛋白水平均升高(P<0.05),RhoAmRNA与蛋白水平降低(P<0.05)。结论:小分子化合物I942结合UVB照射诱导能够增加黑素细胞的树突数量,提高细胞黑色素含量,提升酪氨酸酶活性,促进黑色素合成。

Objective Explore the efiect of small molecule compound I942 on the promotion of human melanocyte dendritic formation and melanin synthesis induced by UVB irradiation.Methods Treatment of immortalized human epidermal melanocyte cell line PIG1 cells with the small molecule compound I942 at a concentration of 0,3.125,6.25,12.5,25,50μmol/L.CCK-8 method to detect cell viability of each group at 24,48,72 h.PIG1 cells are divided into control group,UVB group,UVB+2.5μmol/L I942 group,UVB+5μmol/L I942 group,UVB+10μmol/L I942 group.According to the group name for corresponding processing.Dopa staining to observe the morphology of each group of cells.Immunofluorescence staining to observe the distribution of melanosomes and skeleton protein F-actin.Determination of melanin content by sodium hydroxide pyrolysis method.Determination of tyrosinase activity by DOPA oxidation reaction method.qRT-PCR,WB detection of dendritic morphology-related,melanin synthesis-related mRNA and protein expression levels.Results The small molecule compound I942 at the concentration of 0,3.125,6.25,12.5,25,50μmol/L has no significant efiect on the viability of PIG1 cells(P>0.05).Compared with the control group,the number of cell dendrites,gp100,F-actin,melanin content,tyrosinase activity,MITF,TYR,TRP-1,TRP-2,Rac1 mRNA and protein levels increased in UVB group,UVB+2.5μmol/L I942 group,UVB+5μmol/L I942 group,UVB+10μmol/L I942 group(P<0.05),and RhoA mRNA and protein levels decreased(P<0.05).Conclusion The small molecule compound I942 combined with UVB irradiation can increase the number of dendrites of melanocytes,increase the melanin content of cells,increase tyrosinase activity,and promote melanin synthesis.