Shimizu Y, Kielar D, Masuno H, Minokoshi Y, Shimazu T
Department of Medical Biochemistry, Ehime University School of Medicine.
J Biochem. 1994 Jun;115(6):1069-74. doi: 10.1093/oxfordjournals.jbchem.a124459.
Glucose uptake into brown adipose tissue is enhanced directly by norepinephrine released from the sympathetic nerves. In the present study, we tried to establish culture conditions for brown adipocytes which are favorable for investigation of this unique glucose transport. Stromal-vascular cells isolated from the interscapular brown adipose tissue of newborn rats differentiated into brown adipocytes expressing the uncoupling protein, when the cells were maintained on collagen-coated dishes. These cells, however, did not show an increase in 2-[3H]deoxyglucose transport in response to insulin or norepinephrine, nor did they exhibit expression of the GLUT4 glucose transporter, whereas GLUT1 was present, as judged on Western blotting. Pre-treatment of confluent cells with dexamethasone induced a response of glucose transport to either insulin or norepinephrine, and the expression of GLUT4, together with notable accumulation of lipid droplets. The induction of GLUT4 expression by dexamethasone was dose-dependent and potentiated by insulin. These results indicate that treatment of cultured brown adipocytes with dexamethasone makes it feasible to analyze the mechanism underlying the enhancement of glucose transport induced by norepinephrine.
交感神经释放的去甲肾上腺素可直接增强棕色脂肪组织对葡萄糖的摄取。在本研究中,我们试图建立有利于研究这种独特葡萄糖转运的棕色脂肪细胞培养条件。当从新生大鼠肩胛间棕色脂肪组织分离的基质血管细胞维持在胶原包被的培养皿上时,它们会分化为表达解偶联蛋白的棕色脂肪细胞。然而,这些细胞对胰岛素或去甲肾上腺素刺激的2-[3H]脱氧葡萄糖转运未增加,也未表现出GLUT4葡萄糖转运蛋白的表达,而通过蛋白质免疫印迹法判断,GLUT1是存在的。用皮质酮预处理汇合细胞可诱导葡萄糖转运对胰岛素或去甲肾上腺素产生反应,并诱导GLUT4表达,同时伴有明显的脂滴积累。皮质酮诱导的GLUT4表达呈剂量依赖性,且胰岛素可增强该作用。这些结果表明,用皮质酮处理培养的棕色脂肪细胞使得分析去甲肾上腺素诱导葡萄糖转运增强的潜在机制成为可能。