The phospholipase C inhibitor U73122 attenuates trans-10, cis-12 conjugated linoleic acid-mediated inflammatory signaling and insulin resistance in human adipocytes.
The phospholipase C inhibitor U73122 attenuates trans-10, cis-12 conjugated linoleic acid-mediated inflammatory signaling and insulin resistance in human adipocytes.
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DOI：
10.3945/jn.112.173161
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发表时间：
2013-05
期刊：
影响因子：
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通讯作者：
Wan Shen;K. Martinez;Chia‐Chi Chuang;M. McIntosh
中科院分区：
文献类型：
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作者：
Wan Shen;K. Martinez;Chia‐Chi Chuang;M. McIntosh
我们已经证明，反式-10，顺式-12共轭亚油酸（18：2 t10，c12）介导的人脂肪细胞的脱脂依赖于增加的细胞内钙和激活的炎症信号在人原代脂肪细胞。这些数据与磷脂酶C（PLC）依赖性细胞信号传导的甘油二酯和三磷酸肌醇的作用一致。为了检验PLC是这些细胞对18：2 t10，c12反应的上游激活剂的假设，用1-[6-（（17β-3-甲氧基雌甾-1，3，5（10）-三烯-17-基）氨基）己基]-1H-吡咯-2，5-二酮（U 73122）（一种通用PLC抑制剂）预处理人脂肪细胞的原代培养物，然后进行18：2 t10，c12处理。U 73122可在48 h内减轻18：2 t10，c12介导的胰岛素抵抗，并在18-24 h内抑制过氧化物酶体增殖物激活受体（PPAR）γ、胰岛素刺激的葡萄糖转运蛋白-4、乙酰辅酶A羧化酶-1和硬脂酰辅酶A去饱和酶-1的mRNA水平以及PPARγ的蛋白水平。U 73122在18-24小时内抑制18：2 t10、c12介导的炎症相关基因钙/钙调蛋白依赖性蛋白激酶-β、环加氧酶-2、单核细胞趋化蛋白-1、白细胞介素（IL）-6和IL-8的诱导，抑制IL-6和IL-8的分泌，以及细胞外信号相关激酶、c-Jun N末端激酶和c-Jun的激活。18：2 t10，c12在6-24 h内使热休克蛋白mRNA水平升高，在3 min内使细胞内钙离子浓度升高，而U 73122可抑制热休克蛋白mRNA水平的升高。最后，18：2 t10，c12在3分钟内增加了质膜上PLCγ1的丰度。综上所述，这些数据表明PLC在18：2 t10，c12介导的人原代脂肪细胞胞内钙积累、炎症信号、去脂和胰岛素抵抗的激活中起重要作用。
We have demonstrated that trans-10, cis-12 conjugated linoleic acid (18:2t10,c12)-mediated delipidation of human adipocytes was dependent on increased intracellular calcium and activation of inflammatory signaling in human primary adipocytes. These data are consistent with the actions of diacylglycerol and inositol triphosphate derived from phospholipase C (PLC)-dependent cell signaling. To test the hypothesis that PLC was an upstream activator of these cellular responses to 18:2t10,c12, primary cultures of human adipocytes were pretreated with 1-[6-((17β-3-methoxyestra-1,3,5 (10)-trien-17-yl)amino)hexyl]-1H-pyrrole-2,5-dione (U73122), a universal PLC inhibitor, followed by 18:2t10,c12 treatment. U73122 attenuated 18:2t10,c12-mediated insulin resistance within 48 h and suppression of the mRNA levels of peroxisome proliferator-activated receptor (PPAR)γ, insulin-stimulated glucose transporter-4, acetyl-CoA carboxylase-1, and stearoyl-CoA desaturase-1, and the protein levels of PPARγ within 18-24 h. U73122 inhibited 18:2t10,c12-mediated induction of the inflammatory-related genes calcium/calmodulin-dependent protein kinase-β, cyclooxygenase-2, monocyte chemoattractant protein-1, interleukin (IL)-6, and IL-8, secretion of IL-6 and IL-8, and the activation of extracellular signal-related kinase, c-Jun N-terminal kinase, and c-Jun within 18-24 h. Moreover, 18:2t10,c12 increased the mRNA levels of heat shock proteins within 6-24 h and intracellular calcium concentrations within 3 min, which were inhibited by U73122. Lastly, 18:2t10,c12 increased the abundance of PLCγ1 in the plasma membrane within 3 min. Taken together, these data suggest that PLC plays an important role in 18:2t10,c12-mediated activation of intracellular calcium accumulation, inflammatory signaling, delipidation, and insulin resistance in human primary adipocytes.