Glucocorticoid Signaling Defines a Novel Commitment State during Adipogenesis In Vitro
Glucocorticoid Signaling Defines a Novel Commitment State during Adipogenesis In Vitro
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DOI：
10.1091/mbc.e08-04-0420
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发表时间：
2008-10-01
影响因子：
3.3
通讯作者：
Yamamoto, Keith R.
中科院分区：
文献类型：
--
作者：
Pantoja, Carlos;Huff, Jason T.;Yamamoto, Keith R.
3 T3-L1前脂肪细胞的分化可以通过用含有合成糖皮质激素地塞米松（dex）、胰岛素、磷酸二酯酶抑制剂甲基异丁基黄嘌呤（IBMX）和胎牛血清（FBS）的因子“鸡尾酒”（DIM）2-d处理来诱导。我们暂时解偶联的四个DIM组件的活动，并发现，与dex处理48小时，然后IBMX处理48小时是足够的脂肪形成，而与IBMX处理，然后dex未能诱导显着的分化。用C3 H10 T1/2和原代间充质干细胞获得了类似的结果。通过dex和IBMX顺序处理分化的3 T3-L1脂肪细胞显示与DIM脂肪细胞相当的胰岛素敏感性，但对ISO刺激的脂解和降低的甘油三酯含量具有较低的敏感性。非分化的IBMX-then-dex处理产生脂肪形成转录调节因子C/EBP β和C/EBP δ的瞬时表达，并且几乎不诱导终末分化因子C/EBP β和PPAR γ。此外，脂肪生成抑制剂前脂肪细胞因子-1（Pref-1）被DIM或dex-then-IBMX抑制，但不被IBMX-then-dex处理。我们的结论是，糖皮质激素驱动前脂肪细胞的一种新的中间细胞状态，地塞米松引发的前脂肪细胞，在细胞培养的脂肪形成过程中，Pref-1的抑制可能是前脂肪细胞的细胞命运决定因素。
Differentiation of 3T3-L1 preadipocytes can be induced by a 2-d treatment with a factor "cocktail" (DIM) containing the synthetic glucocorticoid dexamethasone (dex), insulin, the phosphodiesterase inhibitor methylisobutylxanthine (IBMX) and fetal bovine serum (FBS). We temporally uncoupled the activities of the four DIM components and found that treatment with dex for 48 h followed by IBMX treatment for 48 h was sufficient for adipogenesis, whereas treatment with IBMX followed by dex failed to induce significant differentiation. Similar results were obtained with C3H10T1/2 and primary mesenchymal stem cells. The 3T3-L1 adipocytes differentiated by sequential treatment with dex and IBMX displayed insulin sensitivity equivalent to DIM adipocytes, but had lower sensitivity to ISO-stimulated lipolysis and reduced triglyceride content. The nondifferentiating IBMX-then-dex treatment produced transient expression of adipogenic transcriptional regulatory factors C/EBP beta and C/EBP delta, and little induction of terminal differentiation factors C/EBP beta and PPAR gamma. Moreover, the adipogenesis inhibitor preadipocyte factor-1 (Pref-1) was repressed by DIM or by dex-then-IBMX, but not by IBMX-then-dex treatment. We conclude that glucocorticoids drive preadipocytes to a novel intermediate cellular state, the dex-primed preadipocyte, during adipogenesis in cell culture, and that Pref-1 repression may be a cell fate determinant in preadipocytes.