长链非编码RNA在牛FAPs细胞成纤维/成脂诱导分化中的调控机制研究

It was conclusively established in rodents that intramuscular adipocytes and fibroblasts share immediate common ancestor cells, termed fibrogenic/adipogenic progenitor cells—FAPs . These cells are mainly located in the stromal-vascular fraction of skeletal muscle. The FAPs will develop into fibroblasts and adipocytes in fetal ,the numbern and ratio of FAPs commitment to fibroblasts and adipocytes are crucial to the marbling and tenderness of bovin meat.However, the mechnism and the lncRNA-dependent regulatory networks involved in the processes of fibrogenesis and adipogenesis in cattle remain unclear.We will investigate the lncRNA and their potential target genes involved in fibrogenesis and adipogenesis by lncRNA sequencing and bioinformatics analysis.Based on this,the biological function of lncRNA on FAPs differentation will be identified with the over-expression and LNA-expression methods.The potential target genes of lncRNA will be verified by bioinformatics analysis,construction of Sensor-report vector,and Western-blot,then the signaling pathway of the target genes will be elucidated. The results of this project will provide useful information for the interpretation of FAPs differentation mechanism in beef cattle.Also, the work will benefit to enhance the beef quality of Chinese cattle .

FAPs（fibrogenic/adipogenic progenitor cells,FAPs）细胞是位于骨骼肌间质血管附近的一组间充质干细胞，它具有双向分化潜能，既可以向成纤维细胞分化，也可以向脂肪细胞分化，而且向二者分化的比例和数量决定着牛肉的大理石花纹等级和嫩度，但其分化机制尚不明确。本研究拟通过IncRNA测序和生物信息学分析，筛选FAPs向成纤维和脂肪分化过程中差异表达lncRNA，然后通过超表达和抑制表达验证差异表达lncRNA的生物学功能；通过Western blot、Sensor-report载体构建验证差异表达lncRNA靶基因功能，并深入分析靶基因潜在的信号调控网络，初步探讨FAPs向成纤维和脂肪细胞分化的分子调控机制，为进一步提高我国黄牛牛肉品质奠定理论基础。

FAPs（fibrogenic/adipogenic progenitor cells,FAPs）细胞是位于骨骼肌间质血管附近的一组间充质干细胞，它具有双向分化潜能，既可以向成纤维细胞分化，也可以向脂肪细胞分化，而且向二者分化的比例和数量决定着牛肉的大理石花纹等级和嫩度，但其分化机制尚不明确。为了揭示 lncRNA 调控牛FAPs细胞分化的分子机理，本项目以南阳牛背最长肌为素材，分离FAPs并构建了FAPs细胞向脂肪和成纤维分化的cDNA文库，运用第二代高通量测序与生物信息学技术，筛选出了473 条差异 lncRNA （ 313个上调，160个下调 ），同时还发现了143个差异表达circRNA (102个上调，41个下调)。通过差异表达的lncRNA-miRNA-mRNA共表达网络分析发现，bta-miR-2881、bta-miR-177b、bta-miR-177a、bta-miR-1584-5p处于该调控网络的关键节点位置，提示这些miRNA可能与上游的lncRNA和下游mRNA组成调节通路，对FAPs细胞向脂肪和成纤维分化产生调控作用。