Factors affecting the induction of uncoupling protein 1 in C2C12 myogenic cells.
Factors affecting the induction of uncoupling protein 1 in C2C12 myogenic cells.
复制标题
DOI：
10.1016/j.cyto.2022.155936
复制
发表时间：
2022-06
期刊：
影响因子：
3.8
通讯作者：
Takehiro Yamamoto;Zhicheng Diao;Masaru Murakami;Fumie Shimokawa;T. Matsui;M. Funaba
中科院分区：
文献类型：
--
作者：
Takehiro Yamamoto;Zhicheng Diao;Masaru Murakami;Fumie Shimokawa;T. Matsui;M. Funaba
棕色/米色脂肪细胞来源于骨骼肌/平滑肌谱系细胞，通过表达线粒体解偶联蛋白1（UCP 1）消耗多余的能量作为热量。先前的研究表明，PR/SET结构域（PRDM）-16或早期B细胞因子（EBF）-2的强制表达诱导C2 C12肌源性细胞中UCP 1阳性脂肪细胞。在此，我们探索了在不引入外源基因的情况下诱导C2 C12细胞中Ucp 1表达的培养条件。罗格列酮（一种过氧化物酶体增殖物激活受体（PPAR）-γ激动剂）、GW 501516（一种PPARδ激动剂）和骨形态发生蛋白（BMP）-7治疗8天，可有效增加Ucp 1表达，以响应蛋白激酶A通路激活剂毛喉素治疗。在罗格列酮和GW 501516存在下，BMP 7剂量依赖性地增加forskolin诱导的Ucp 1表达;然而，Ucp 1诱导不需要GW 501516。此外，结构相关的蛋白质，BMP 6和BMP 9，有效地增加了罗格列酮处理的细胞中毛喉素诱导的Ucp 1表达。UCP 1蛋白定位于细胞与脂滴，但脂肪细胞并不总是阳性的UCP 1。需要持续用BMP 7处理才能有效诱导毛喉素处理的Ucp 1。Prdm 16的显著表达未被检测到，与治疗无关，并且用罗格列酮、GW 501516和BMP 7治疗不影响Ebf 2的表达水平。在罗格列酮处理的细胞中，BMP 9增加了成纤维细胞生长因子受体（Fgfr）-3的表达水平，上调Fgfr 3转录的分子与刺激Ucp 1转录的分子部分重叠。本研究结果为肌源性细胞向棕色脂肪细胞的实际分化提供了基础信息。
Brown/beige adipocytes, which are derived from skeletal muscle/smooth muscle-lineage cells, consume excess energy as heat through the expression of mitochondrial uncoupling protein 1 (UCP1). Previous studies have shown that forced expression of PR/SET domain (PRDM)-16 or early B-cell factor (EBF)-2 induced UCP1-positive adipocytes in C2C12 myogenic cells. Here, we explored the culture conditions to induceUcp1expression in C2C12 cells without introducing exogenous genes. Treatment with rosiglitazone (a peroxisome proliferator-activated receptor (PPAR)-γ agonist), GW501516 (a PPARδ agonist), and bone morphogenetic protein (BMP)-7 for 8 days efficiently increasedUcp1expression in response to treatment with forskolin, an activator of the protein kinase A pathway. BMP7 dose-dependently increased forskolin-inducedUcp1expression in the presence of rosiglitazone and GW501516; however, GW501516 was not required forUcp1induction. Additionally, the structurally related proteins, BMP6 and BMP9, efficiently increased forskolin-inducedUcp1expression in rosiglitazone-treated cells. UCP1 protein was localized in cells with lipid droplets, but adipocytes were not always positive for UCP1. Continuous treatment with BMP7 was needed for the efficient induction ofUcp1by forskolin treatment. Significant expression ofPrdm16was not detected, irrespective of the treatment, and treatment with rosiglitazone, GW501516, and BMP7 did not affect the expression levels ofEbf2. Fibroblast growth factor receptor (Fgfr)-3expression levels were increased by BMP9 in rosiglitazone-treated cells, and molecules that upregulateFgfr3transcription partly overlapped with those that stimulateUcp1transcription. The present results provide basic information on the practical differentiation of myogenic cells to brown adipocytes.