Lund S, Vestergaard H, Andersen P H, Schmitz O, Gøtzsche L B, Pedersen O
Division of Endocrinology and Metabolism, University Clinic of Internal Medicine, Aarhus Amtssygehus, Denmark.
Am J Physiol. 1993 Dec;265(6 Pt 1):E889-97. doi: 10.1152/ajpendo.1993.265.6.E889.
The abundance of GLUT-4 protein in both total crude membrane and plasma membrane fractions of vastus lateralis muscle from 13 obese non-insulin-dependent diabetes mellitus (NIDDM) patients and 14 healthy subjects were examined in the fasting state and after supraphysiological hyperinsulinemia. In the basal state the immunoreactive mass of GLUT-4 protein both in the crude membrane preparation and in the plasma membrane fraction was similar in NIDDM patients and control subjects. Moreover, in vivo insulin exposure neither for 30 min nor for 4 h had any impact on the content of GLUT-4 protein in plasma membranes. With the use of the same methodology, antibody, and achieving the same degree of plasma membrane purification and recovery, we found, however, that intraperitoneal administration of insulin to 7-wk-old rats within 30 min increased the content of GLUT-4 protein more than twofold (P < 0.01) in the plasma membrane from red gastrocnemius and soleus muscle. In conclusion, when the subcellular fractionation method was applied to human muscle biopsies taken in the basal state, no difference could be found in the plasma membrane content of immunoreactive GLUT-4 protein between NIDDM patients and normal subjects. With this technique, we were unable to show evidence for a regulatory effect of insulin on the plasma membrane level of GLUT-4 protein in human muscle.
对13名肥胖型非胰岛素依赖型糖尿病(NIDDM)患者和14名健康受试者股外侧肌的总粗膜和质膜部分中GLUT-4蛋白的丰度进行了空腹状态及超生理高胰岛素血症后的检测。在基础状态下,NIDDM患者和对照受试者粗膜制剂和质膜部分中GLUT-4蛋白的免疫反应性质量相似。此外,体内胰岛素暴露30分钟或4小时对质膜中GLUT-4蛋白的含量均无影响。然而,使用相同的方法、抗体并达到相同程度的质膜纯化和回收率,我们发现,在30分钟内向7周龄大鼠腹腔注射胰岛素,可使红腓肠肌和比目鱼肌质膜中GLUT-4蛋白的含量增加两倍多(P<0.01)。总之,当将亚细胞分级分离方法应用于基础状态下采集的人体肌肉活检样本时,NIDDM患者和正常受试者之间质膜中免疫反应性GLUT-4蛋白的含量未发现差异。采用该技术,我们无法证明胰岛素对人体肌肉中GLUT-4蛋白质膜水平有调节作用。