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miR-10a对牦牛脂肪前体细胞增殖与分化的影响

Effect of miR-10a on the proliferation and differentiation of yak adipocyte precursors.

作者信息

Su Quyangangmao, Gao Zhanhong, Zhang Fengshuo, Wu Zhenling, Ji Qiurong, Zhu Kaina, Gui Linsheng

机构信息

College of Agriculture and Animal Husbandry, Qinghai University, Qinghai Province, Xining, 810016, People's Republic of China.

出版信息

J Appl Genet. 2025 May;66(2):435-447. doi: 10.1007/s13353-024-00932-6. Epub 2024 Dec 24.

Abstract

The fat content of yak meat is significantly correlated with the meat quality, and an appropriate fat content helps to improve the texture of the meat. The involvement of miR-10a in regulating the differentiation and proliferation of various cell types has been reported. Therefore, in this study, the effects of miR-10a on lipid droplet accumulation were investigated by transfection of yak adipocyte precursors with an miR-10a inhibitor, followed by Oil Red O, BODIPY, EdU staining, and cell cycle analysis of the transfected and control cells. The relative expression of lipogenic marker genes was determined by RT-qPCR to clarify the effect of miR-10a on the differentiation and proliferation of yak adipocyte precursors. Mature adipocytes were collected for transcriptome analysis to identify differentially expressed target genes and the association of these genes with adipogenic pathways was investigated by GO and KEGG enrichment analyses. In addition, the phylogeny and expression profiles of miR-10a were analyzed in various yak tissues. The results showed that miR-10a could inhibit the differentiation and promote the proliferation of yak adipocyte precursors. Analysis of the RNA-Seq results showed that miR-10a inhibitor and inhibitor NC had six differentially expressed genes: FABP4, AKR1B7, IGF2, ROCK1, IFNB1, and PLA2G3. These genes were found to be involved in the regulation of adipogenesis, with IGF2 and IFNB1 being upregulated in the PI3K-Akt signaling pathway, which is activated upon stimulation by IGF2 and IFNB1 and inhibits the differentiation and promotes the proliferation of yak adipocytes precursor, which in turn affected adipogenesis. Moreover, phylogenetic analysis indicated that miR-10a evolved relatively recently in yak and sheep, while tissue expression profiles showed that miR-10a was highly expressed in yak lung tissues.

摘要

牦牛肉的脂肪含量与肉质显著相关,适当的脂肪含量有助于改善肉质。已有报道称miR-10a参与调节多种细胞类型的分化和增殖。因此,在本研究中,通过用miR-10a抑制剂转染牦牛脂肪细胞前体,随后进行油红O、BODIPY、EdU染色以及对转染细胞和对照细胞进行细胞周期分析,研究了miR-10a对脂滴积累的影响。通过RT-qPCR测定脂肪生成标记基因的相对表达,以阐明miR-10a对牦牛脂肪细胞前体分化和增殖的影响。收集成熟脂肪细胞进行转录组分析,以鉴定差异表达的靶基因,并通过GO和KEGG富集分析研究这些基因与脂肪生成途径的关联。此外,还分析了miR-10a在牦牛各组织中的系统发育和表达谱。结果表明,miR-10a可抑制牦牛脂肪细胞前体的分化并促进其增殖。RNA-Seq结果分析显示,miR-10a抑制剂和抑制剂阴性对照有6个差异表达基因:FABP4、AKR1B7、IGF2、ROCK1、IFNB1和PLA2G3。发现这些基因参与脂肪生成的调控,其中IGF2和IFNB1在PI3K-Akt信号通路中上调,该信号通路在受到IGF2和IFNB1刺激后被激活,抑制牦牛脂肪细胞前体的分化并促进其增殖,进而影响脂肪生成。此外,系统发育分析表明,miR-10a在牦牛和绵羊中进化相对较晚,并通过组织表达谱分析发现miR-10a在牦牛肺组织中高表达。

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