RNAi对羊驼黑色素细胞中MC1R表达量及黑色素合成的影响

Effect of RNAi on MC1R Expression and Melanin Synthesis in the Melanocytes of Alpaca

旨在研究RNAi对羊驼黑色素细胞中MC1R表达量及黑色素合成的影响。合成3对MC1RsiRNA,将MC1RsiRNA瞬时转染羊驼黑色素细胞,qRT-PCR法检测MC1R mRNA表达量,细胞免疫组化技术对MC1R进行定位,利用吸光度值检测黑色素细胞中黑色素的合成量。结果显示,siRNA2组和siRNA3组的MC1R mRNA相对表达量极显著低于空白对照组(P<0.01),siRNA1组和阴性对照组与空白对照组相比差异不显著(P>0.05),siRNA2、siRNA3两组的黑色素合成量低于空白对照组,差异极显著(P<0.01)。将MC1RsiRNA成功转染到羊驼黑色素细胞中,筛选出有效MC1RsiRNA,MC1R的抑制情况与黑色素合成量一致,说明黑色素细胞中MC1R对黑色素的合成具有调控作用,其作用机制需进一步研究。

The objectives of this study were to evaluate the effects of RNAi on MC1 Rexpression and melanin synthesis in the melanocytes of alpaca.Three pairs of specific siRNA for MC1 Rgene were designed for RNA interference in the melanocytes of alpaca.The expression of MC1 RsiRNA was detected by using qRT-PCR technology.The location of MC1 Rwas detected by immunohistochemistry.The amount of melanin synthesis in melanocytes was detected by the absorbance value.The results showed that the relative expression of MC1 R mRNA in the siRNA2 and siRNA3groups was significantly lower(P<0.01)than that in the blank control group,but there were no significant differences at the relative expression of MC1 RmRNA in the siRNA1 and negative control group(P>0.05);the production of melanin synthesis of siRNA2 and siRNA3groups were significantly lower than that in the blank control group(P<0.01).The study was successfully transfected MC1 RsiRNA into alpaca melanocytes and the effective MC1 RsiRNA was screened from 3groups of specific siRNAs.The inhibition of MC1 Ris consistent with the amount of melanin synthesis.MC1 Rof melanocytes played a regulatory role in the synthesis of melanin,and the mechanism need further research.