Brown adipose tissue (BAT) is a specialized tissue in mammals related to thermogenesis. The Astragalus polysaccharide (APS) is the major natural active component of Astragalus membranaceus, which has been recognized as one of the most popular herbal medicines worldwide. The role and possible mechanisms of APS on brown adipocytes differentiation is not well defined. Here, we explored the effect of APS on the differentiation of brown adipocytes in C3H10T 1/2 cells. The results showed that APS promoted the differentiation of brown adipocytes and improved insulin sensitivity along with significant increases in the expression of brown adipogenic marker proteins (C/EBPα, C/EBPβ, and PPARγ), thermogenesis marker proteins (UCP1, PRDM16, and PGC-1α), and insulin sensitivity marker protein (GLUT4). Meanwhile, the results showed that the amount of the phosphorylation of insulin receptor substrate 1 (p-IRS1) and phospho-AKT (p-AKT) which are critical factors in the insulin signaling pathway was increased without changing the total amount of IRS and AKT. Furthermore, the results of RNA-seq showed that APS altered the expression profiles of various miRNAs, and among which the expression of miR-6911 as a universal regulatory factor was significantly decreased. Importantly, we found that miR-6911 regulated the differentiation of brown adipocytes by targeting PR domain-containing 16 (Prdm16). In addition, after transfection of miR-6911 mimics, compared with the control and inhibitor group, PRDM16 protein expression significantly decreased, which was accompanied by the decrease of PPARγ, UCP1, and PGC-1α. Collectively, our results indicated that APS regulated brown adipocytes differentiation in C3H10T 1/2 cells via miRNA-6911 targeting Prdm16.

中文翻译：

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黄芪多糖通过靶向Prdm16的miR-6911调节棕色脂肪细胞分化

棕色脂肪组织 (BAT) 是哺乳动物中与产热相关的一种特殊组织。黄芪多糖（APS）是黄芪的主要天然活性成分，它已被公认为世界上最受欢迎的草药之一。APS 对棕色脂肪细胞分化的作用和可能的机制尚不明确。在这里，我们探讨了 APS 对 C3H10T 1/2 细胞中棕色脂肪细胞分化的影响。结果表明，APS促进棕色脂肪细胞的分化并提高胰岛素敏感性，同时棕色脂肪形成标记蛋白（C/EBPα、C/EBPβ和PPARγ）、产热标记蛋白（UCP1、PRDM16和PGC）的表达显着增加。 -1α) 和胰岛素敏感性标记蛋白 (GLUT4)。同时，结果表明，在不改变 IRS 和 AKT 总量的情况下，胰岛素受体底物 1 (p-IRS1) 和磷酸化 AKT (p-AKT) 的磷酸化量增加，它们是胰岛素信号通路中的关键因素。此外，RNA-seq 的结果表明，APS 改变了各种 miRNA 的表达谱，其中作为通用调节因子的 miR-6911 的表达显着降低。重要的是，我们发现 miR-6911 通过靶向调控棕色脂肪细胞的分化PR 域包含 16 ( Prdm16 )。此外，转染 miR-6911 mimics 后，与对照组和抑制剂组相比，PRDM16 蛋白表达显着降低，并伴有 PPARγ、UCP1 和 PGC-1α 的降低。总的来说，我们的结果表明 APS 通过靶向Prdm16的 miRNA-6911 调节 C3H10T 1/2 细胞中棕色脂肪细胞的分化。