【摘要】 为深入探讨抑制miR-99b-3p对山羊骨骼肌细胞发育的影响机制,建立了地塞米松诱导的山羊骨骼肌卫星细胞(Skeletal muscle satellite cells,SMSCs)应激模型并转染miR-99b-3p,并通过转录组测序研究抑制miR-99b-3p对SMSCs的分子调控机制。结果表明:转染miR-99b-3p可促进SMSCs细胞增殖以及肌肉分化基因MyoG和MyoD的表达,抑制miR-99b-3p则会抑制细胞增殖和分化基因表达,并导致细胞中761个基因差异表达。KEGG分析显示,差异基因大多富集于与疾病相关的通路;GO富集分析表明,差异基因主要参与细胞增殖和DNA修复相关生物学过程。此外,鉴定到17个差异表达的miRNAs,包含9个上调和8个下调miRNAs。对差异miRNA靶基因进行预测分析,并与差异基因比对,筛选到18个重叠的差异基因,这些差异基因主要富集于与致病相关的通路。本研究可为阐明断奶应激对山羊生长发育的分子调控机制提供理论依据。更多还原

【Abstract】 To explore the mechanism of inhibiting miR-99b-3p on the development of goat skeletal muscle cells,a dexamethasone induced stress model of goat skeletal muscle satellite cells(SMSCs) was first established and miR-99b-3p was transfected.Then,the molecular regulatory mechanism of miR-99b-3p inhibition on SMSCs was studied by transcriptome sequencing.The results showed that the transfection of miR-99b-3p promoted the proliferation of SMSCs and the expression of muscle differentiation gyenes MyoG and MyoaD,while inhibition of miR-99b-3p inhibited cell proliferation and differentiation genes expression.Inhibition of miR-99b-3p resulted in differential expression of 761 genes in cells;KEGG analysis showed that most differentially expressed genes were enriched in pathways related to diseases;GO enrichment analysis showed that differentially expressed genes were mainly involved in biological processes related to cell proliferation and DNA repair.In addition,17 differentially expressed miRNAs were identified,including 9 upregulated and 8 downregulated miRNAs.By predicting and analyzing differentially expressed miRNA target genes,and comparing them with differentially expressed genes,a total of 18 overlapping differentially expressed genes were screened,which were mainly enriched in pathways related to disease.This study provides a theoretical basis for elucidating the molecular regulatory mechanism of weaning stress on goat growth and development.更多还原