Tao Y, Cianflone K, Sniderman A D, Colby-Germinario S P, Germinario R J
McGill Unit for the Prevention of Cardiovascular Disease, Royal Victoria Hospital, Montreal, Que., Canada.
Biochim Biophys Acta. 1997 Feb 18;1344(3):221-9. doi: 10.1016/s0005-2760(96)00144-0.
Acylation-stimulating protein (ASP), a human plasma protein, is a potent stimulator of triglyceride synthesis and glucose transport in both human adipocytes and fibroblasts. The purpose of the present in vitro study was to examine the effect of ASP on glucose transport in muscle cells. ASP stimulated 2-deoxy-glucose transport (2-DG) in differentiated rat L6 myotubes in a time (30 min to 24 h) and concentration dependent manner (97% increase). The magnitude of the ASP effect on glucose transport was comparable to the time- and concentration-dependent effects seen with insulin (125% increase), but was additive to insulin, pointing to involvement of differential signalling pathways. ASP stimulation was dependent on cell differentiation in that glucose transport increased by only 12% in myoblasts, comparable to the effect of insulin in myoblasts (15% increase) demonstrating selective responsiveness of the differentiated myotubes to ASP and insulin. The mechanism for the ASP induced increase in glucose transport was also examined. ASP increased the Vmax for 2-DG transport by 183% (4.02 vs. 1.42 nmol/mg cell protein/30 s; ASP vs. Control, respectively). This could be explained by an increased translocation of glucose transporters (GLUT 1, GLUT 4 and GLUT 3) to the plasma membrane surface as demonstrated by Western analysis (+43% P < 0.05, +30% P < 0.05, and +49% P < 0.05, respectively). The effects of ASP were equal to those of insulin (+47%, +26% and +53% for GLUT 1, GLUT 4 and GLUT 3, respectively) and in all cases were paralleled by comparable glucose transport increases under the same incubation conditions. After long-term stimulation (24 h), Western analysis indicated that ASP had a permissive effect on insulin stimulated increases in total GLUT3 and GLUT4 cellular transporter content. These results suggest that muscle is also responsive to ASP and that ASP may play a role in glucose metabolism in both muscle and adipose tissue.
酰化刺激蛋白(ASP)是一种人体血浆蛋白,在人体脂肪细胞和成纤维细胞中,它是甘油三酯合成和葡萄糖转运的有效刺激物。本体外研究的目的是检测ASP对肌肉细胞中葡萄糖转运的影响。ASP以时间(30分钟至24小时)和浓度依赖方式(增加97%)刺激分化的大鼠L6肌管中的2-脱氧葡萄糖转运(2-DG)。ASP对葡萄糖转运的作用强度与胰岛素的时间和浓度依赖性作用相当(增加125%),但与胰岛素的作用具有相加性,表明涉及不同的信号通路。ASP刺激依赖于细胞分化,因为成肌细胞中葡萄糖转运仅增加12%,与胰岛素在成肌细胞中的作用相当(增加15%),这表明分化的肌管对ASP和胰岛素具有选择性反应。还研究了ASP诱导葡萄糖转运增加的机制。ASP使2-DG转运的Vmax增加了183%(分别为4.02对1.42 nmol/mg细胞蛋白/30秒;ASP对对照组)。这可以通过葡萄糖转运蛋白(GLUT 1、GLUT 4和GLUT 3)向质膜表面的转位增加来解释,如蛋白质免疫印迹分析所示(分别增加43%,P < 0.05;增加30%,P < 0.05;增加49%,P < 0.05)。ASP的作用与胰岛素的作用相当(GLUT 1、GLUT 4和GLUT 3分别增加47%、26%和53%),并且在所有情况下,在相同孵育条件下葡萄糖转运的增加与之平行。长期刺激(24小时)后,蛋白质免疫印迹分析表明,ASP对胰岛素刺激的总GLUT3和GLUT4细胞转运蛋白含量增加具有允许作用。这些结果表明,肌肉对ASP也有反应,并且ASP可能在肌肉和脂肪组织的葡萄糖代谢中发挥作用。