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本实验旨在研究miR-18b-3p及其靶基因SCD调控绵羊前体脂肪细胞分化的作用机制。采集3日龄公羊背最长肌,培养绵羊前体脂肪细胞并进行诱导分化,采用油红O染色检测细胞分化情况;通过TargetScan预测miR-18b-3p的靶基因,通过qRT-PCR测定miR-18b-3p、靶基因SCD以及分化标志基因PPARγ、CEBPα在分化过程中的时序性表达;分别转染miR-18b-3p过表达载体(mimics)或抑制载体(inhibitor)及阴性对照至前体脂肪细胞后,通过qRT-PCR及Western blot检测miR-18b-3p对SCD以及分化标志基因表达量的作用;采用双荧光素酶证明miR-18b-3p是否靶向SCD基因。结果表明:miR-18b-3p在分化过程中的表达量先上升后下降,在第6天表达量最高;SCD表达量低于阴性对照组(P<0.01),PPARγ及CEBPα基因表达量均低于阴性对照组(P<0.05),SCD蛋白表达量低于阴性对照组(P<0.01),油红O染色面积减少(P<0.05);转染miR-18b-3p inhibitor后,SCD、CEBPα基因表达量均高于阴性对照组(P<0.01),PPARγ基因表达量高于阴性对照组(P<0.05),SCD蛋白表达量高于阴性对照组(P<0.01),油红O染色面积增加(P<0.05);miR-18b-3p对SCD基因具有直接靶向作用。本实验首次在绵羊上证明miR-18b-3p可以通过靶向SCD抑制前体脂肪细胞分化,为绵羊选育给予分子育种技术支撑。
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基本信息:
DOI：10.19556/j.0258-7033.20210306-05
中图分类号:S826
引用信息:
[1]王家麒,韩福慧,赵乐等.miR-18b-3p靶向SCD调控绵羊前体脂肪细胞分化的研究[J].中国畜牧杂志,2021,57(05):117-123.DOI:10.19556/j.0258-7033.20210306-05.
基金信息:
国家绒毛用羊产业技术体系专项资金（CARS-39-05）