Department of Animal Science, University of Manitoba, 201 Animal Science building, Winnipeg, MB R3T 2N2, Canada.
Department of Poultry Science, University of Georgia, 303 Poultry Science building, Athens, GA 30602-2772, USA.
Int J Mol Sci. 2020 Jan 9;21(2):412. doi: 10.3390/ijms21020412.
Understanding of adipogenesis is important to find remedies for obesity and related disorders. In addition, it is also critical in bone disorders because there is a reciprocal relationship between adipogenesis and osteogenesis in bone micro-environment. Oxysterols are pro-osteogenic and anti-adipogenic molecules via hedgehog activation in pluripotent bone marrow stomal cells. However, no study has evaluated the role of specific oxysterols in C3H10T1/2 cells, which are a good cell model for studying osteogenesis and adipogenesis in bone-marrows. Thus, we investigated the effects of specific oxysterols on adipogenesis and expression of adipogenic transcripts in C3H10T1/2 cells. Treatment of cells with DMITro significantly induced mRNA expression of Pparγ. This induction was significantly inhibited by 25-HC. The expression of , and was also inhibited by 25-HC. To determine the mechanism by which 25-HC inhibits adipogenesis, the effects of the hedgehog signalling pathway inhibitor, cyclopamine and CUR61414, were evaluated. Treatment of C3H10T1/2 cells with DMITro + cyclopamine or DMITro + CUR61414 for 96h did not modulate adipocyte differentiation; cyclopamine and CUR61414 did not reverse the inhibitory effects of 25-HC, suggesting that the canonical hedgehog signalling may not play a role in the anti-adipogenic effects of 25-HC in C3H10T1/2 cells. In addition, LXR agonist did not inhibit adipogenesis, but 25-HC strongly inhibits adipogenesis of C3H10T1/2 cells. Our observations showed that 25-HC was the most potent oxysterol in inhibiting adipogenesis and the expression of key adipogenic transcripts in C3H10T1/2 cells among the tested oxysterols, suggesting its potential application in providing an intervention in osteoporosis and obesity. We also report that the inhibitory effects of 25-HC on adipogenic differentiation in C3H10T1/2 cells are not mediated by hedgehog signaling and LXR.
了解脂肪生成对于寻找肥胖症和相关疾病的治疗方法非常重要。此外，在骨疾病中，脂肪生成和骨生成之间存在骨微环境中的相互关系，因此这一点也至关重要。在多能骨髓基质细胞中，通过 hedgehog 激活，氧化固醇是促成骨和抗脂肪生成的分子。然而，在 C3H10T1/2 细胞中，尚未有研究评估特定氧化固醇的作用，C3H10T1/2 细胞是研究骨髓中成骨和脂肪生成的良好细胞模型。因此，我们研究了特定氧化固醇对 C3H10T1/2 细胞脂肪生成和脂肪生成转录物表达的影响。细胞用 DMITro 处理可显著诱导 PparγmRNA 的表达。这种诱导作用被 25-HC 显著抑制。25-HC 还抑制了 、 和 的表达。为了确定 25-HC 抑制脂肪生成的机制，评估了 hedgehog 信号通路抑制剂环巴胺和 CUR61414 的作用。用 DMITro+环巴胺或 DMITro+CUR61414 处理 C3H10T1/2 细胞 96 小时不会调节脂肪细胞分化；环巴胺和 CUR61414 没有逆转 25-HC 的抑制作用，表明 canonical hedgehog 信号通路可能没有在 25-HC 对 C3H10T1/2 细胞的抗脂肪生成作用中发挥作用。此外，LXR 激动剂不会抑制脂肪生成，但 25-HC 强烈抑制 C3H10T1/2 细胞的脂肪生成。我们的观察结果表明，在测试的氧化固醇中，25-HC 是抑制 C3H10T1/2 细胞脂肪生成和关键脂肪生成转录物表达最有效的氧化固醇，这表明其在提供骨质疏松症和肥胖症干预方面具有潜在应用。我们还报告说，25-HC 对 C3H10T1/2 细胞脂肪生成分化的抑制作用不是通过 hedgehog 信号和 LXR 介导的。
