非转移性黑色素瘤糖蛋白B对小鼠黑色素细胞色素生成的影响

摘要

旨在探索非转移性黑色素瘤糖蛋白B (GPNMB)在不同毛色小鼠皮肤是否存在差异性表达,确定其对黑色素生成的影响.随机选取C57BL/6J品系出生12d的黑、棕、灰小鼠各3只,背部剪毛后,每只小鼠各取3块皮肤组织,采用免疫组织化学检测,结果表明,GPNMB在不同毛色小鼠皮肤的毛基质、内外毛根鞘、毛乳头等区域均有表达.利用Western blot和qRT-PCR方法分析,结果显示,灰色小鼠皮肤中GPNMB表达量最低.过表达GPNMB后,与空载体组相比,MC1R、β-catenin和SLC45 A2相对表达量显著升高;用紫外线B(UVB)处理黑色素细胞,同时检测GPNMB和MC1R的表达量变化.结果表明,UVB对GPNMB和MC1R有影响,并且不同剂量UVB辐射对GPNMB影响不同.综上表明,GPNMB在不同毛色小鼠皮肤存在差异性表达,并且通过影响黑色素细胞色素的生成进而参与毛色的生成.%This study aims to investigate whether GPNMB differentially expresses in the skin of mice with various hair colors and to determine its effect on the melanin synthesis.Three approximately 12-day-old healthy C57BL/6 mice were randomly selected from strains with black,brown and gray coat colors,respectively.Three pieces of dorsal skin tissue were collected from each mouse using a skin collector after shaving.Immunohistochemistry assay revealed that GPNMB was mainly located in the hair follicle matrix,the inner and outer root sheath,and the dermal papilla in the skin with various hair colors.The Western blot analysis and qRT-PCR assay showed that the expression level of GPNMB was the lowest in the skin of gray mice.Compared with the control group,relative levels of MC1R,β-catenin and SLC45A2 significantly increased in mouse melanocytes overexpressing GPNMB.we detected the levels of GPNMB and MC1R in mouse melanocytes which had been radiated by UVB.The results showed that UVB had an impact on the GPNMB and MC1R expression,and the level of GPNMB was various with different UVB radiation doses.The above results suggested that GPNMB was differentially expressed in the skin of mice with different hair colors,and it was involved in the formation of the hair color by regulating the melanin synthesis.