内皮素3对小鼠黑色素沉着相关基因表达的影响

Effects of EDN3 on the Expression of Melanin-related Genes in Mouse Melanocytes

试验旨在探究内皮素3(endothelin 3,EDN3)在不同毛囊时期小鼠皮肤中是否存在差异表达及其对黑色素细胞黑色素沉着相关基因表达的影响。通过RT-PCR技术对EDN3、EDNRB在不同毛囊时期小鼠皮肤的表达情况进行定量分析发现,EDN3和EDNRB在小鼠不同毛囊时期皮肤样品中均有表达,在毛囊生长初期和中期小鼠皮肤中EDN3 mRNA表达量是末期的2.18倍(P<0.05)和1.15倍(P>0.05),毛囊生长初期和中期EDNRB mRNA表达量是末期的16.8倍(P<0.01)和9.9倍(P<0.01)。为了进一步揭示EDN3在黑色素细胞色素沉着中的重要作用,通过细胞转染技术使小鼠黑色素细胞过表达EDN3并测定其黑色素含量及色素沉着相关基因的表达水平发现,与空载组相比,体外转染EDN3的黑色素细胞黑色素含量明显增加。此外,MITF mRNA显著降低(P<0.05);TYR、TYRP2、EDNRB、c-Kit和EDN1 mRNA显著升高2.04倍(P<0.05)、1.44倍(P<0.05)、1.41倍(P<0.05)、5.21倍(P<0.01)和3.27倍(P<0.01)。MITF蛋白水平显著降低(P<0.05),TYR、TYRP2、EDNRB和c-Kit蛋白水平显著升高1.48倍(P<0.01)、4.61倍(P<0.01)、1.27倍(P<0.05)和2.64倍(P<0.01)。综上所述,EDN3在不同毛囊时期小鼠皮肤中均可有效表达,且表达量存在显著差异。黑色素细胞中过量表达EDN3,使色素沉着相关基因的表达量及黑色素含量增加。

The study was conducted to explore whether endothelin 3（EDN3）could expressed in mouse skin of different hair follicle period and its influence on the expression of melanin-related genes.The expression of EDN3 and EDNRB in mouse skin of different hair follicle period were analyzed by RT-PCR technology.The results showed that EDN3 and EDNRB were both expressed in mouse skin samples in different hair follicle period,and the relative expression of EDN3 mRNA in anagen and catagen were 2.18 times（P〈0.05）and 1.15 times（P〉0.05）as much as telogen,respectively,while the EDNRB mRNA in anagen and catagen were 16.8 times（P〈0.01）and 9.9 times（P〈0.01）,respectively.In order to further reveal the important role of EDN3 in pigmentation,the EDN3 were overexpressed in mice melanocytes by cell transfection technique,the melanin content of melanocytes and the relative expression levels of pigmentation-related genes were tested.The results showed that the melanin content was obviously increased after transfecting EDN3 into mice melanocytes in vitro.Furthermore,MITF mRNA was decreased（P〈0.05）,TYR,TYRP2,EDNRB,c-Kit and EDN1 mRNA were significantly increased 2.04 times（P〈0.05）,1.44 times（P〈0.05）,1.41 times（P〈0.05）,5.21 times（P〈0.01）and3.27 times（P〈0.01）.The expression of MITF protein was significantly decreased（P〈0.05）;TYR,TYRP2,c-Kit and EDNRB proteins were significantly increased to 1.48 times（P〈0.01）,4.61 times（P〈0.01）,1.27 times（P〈0.05）and 2.64 times（P〈0.01）,respectively.In conclusion,EDN3 was effectively expressed in mouse skin of different hair follicle period,and its expression levels had significant differences,and overexpressing EDN3 could increase the expression level of melanin-related genes and melanin content.