Comparative Proteomic Study of Fatty Acid-treated Myoblasts Reveals Role of Cox-2 in Palmitate-induced Insulin Resistance.
Comparative Proteomic Study of Fatty Acid-treated Myoblasts Reveals Role of Cox-2 in Palmitate-induced Insulin Resistance.
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脂肪酸处理的成肌细胞的比较蛋白质组学研究揭示了 Cox-2 在棕榈酸酯诱导的胰岛素抵抗中的作用
DOI：
10.1038/srep21454
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发表时间：
2016-02-22
影响因子：
4.6
通讯作者：
Liu P
中科院分区：
文献类型：
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作者：
Chen X;Xu S;Wei S;Deng Y;Li Y;Yang F;Liu P
大量研究表明，饱和脂肪酸(FA)如棕榈酸(PA)可抑制骨骼肌细胞中的胰岛素信号传导，而油酸(OA)等单不饱和脂肪酸可逆转PA对胰岛素信号传导的影响。这些相反作用的详细分子机制仍然难以捉摸。在这里，我们提供了骨骼肌成肌细胞系C2C12的比较蛋白质组学研究，这些细胞系未处理或经PA处理，以及PA+OA处理。在这项研究中，共有3437个蛋白质被SILAC定量，其中29个蛋白质属于OA逆转PA效应的模式。用qRT-PCR和Western印迹方法验证了部分蛋白的表达。最显著的变化是环氧合酶-2(COX-2)。除了全细胞比较蛋白质组学研究外，我们还比较了脂滴(LD)相关蛋白，并确定COX-2是FA处理下的三个主要变化蛋白之一。这一发现随后得到了免疫荧光的证实。最后，COX-2选择性抑制剂塞来昔布保护细胞免受PA降低的胰岛素信号Akt磷酸化的影响。综上所述，这些结果不仅提供了FA处理过程中蛋白质表达变化的数据集，而且表明COX-2和脂滴(LDS)在PA和OA介导的细胞过程中是潜在的参与者。
Accumulated studies demonstrate that saturated fatty acids (FAs) such as palmitic acid (PA) inhibit insulin signaling in skeletal muscle cells and monounsaturated fatty acids such as oleic acid (OA) reverse the effect of PA on insulin signaling. The detailed molecular mechanism of these opposite effects remains elusive. Here we provide a comparative proteomic study of skeletal myoblast cell line C2C12 that were untreated or treated with PA, and PA plus OA. A total of 3437 proteins were quantified using SILAC in this study and 29 proteins fall into the pattern that OA reverses PA effect. Expression of some these proteins were verified using qRT-PCR and Western blot. The most significant change was cyclooxygenase-2 (Cox-2). In addition to whole cell comparative proteomic study, we also compared lipid droplet (LD)-associated proteins and identified that Cox-2 was one of three major altered proteins under the FA treatment. This finding was then confirmed using immunofluorescence. Finally, Cox-2 selective inhibitor, celecoxib protected cells from PA-reduced insulin signaling Akt phosphorylation. Together, these results not only provide a dataset of protein expression change in FA treatment but also suggest that Cox-2 and lipid droplets (LDs) are potential players in PA- and OA-mediated cellular processes.