Chen L, Alam T, Johnson J H, Hughes S, Newgard C B, Unger R H
Center for Diabetes Research, Gifford Laboratories, University of Texas Southwestern Medical Center, Dallas 75235.
Proc Natl Acad Sci U S A. 1990 Jun;87(11):4088-92. doi: 10.1073/pnas.87.11.4088.
It has been postulated that a glucose transporter of beta cells (GLUT-2) may be important in glucose-stimulated insulin secretion. To determine whether this transporter is constitutively expressed or regulated, we subjected conscious unrestrained Wistar rats to perturbations in glucose homeostasis and quantitated beta-cell GLUT-2 mRNA by in situ hybridization. After 3 hr of hypoglycemia (glucose at 29 +/- 5 mg/dl), GLUT-2 and proinsulin mRNA signal densities were reduced by 25% of the level in control rats. After 4 days (blood glucose at 57 +/- 7 mg/dl vs. 120 +/- 10 mg/dl in saline-infused control rats), GLUT-2 and proinsulin mRNA densities were reduced by 85% and 65%, respectively (P = 0.001). After 12 days (glucose at 54 +/- 8 mg/dl), GLUT-2 mRNA signal density was undetectable whereas proinsulin mRNA was reduced by 51%. After 12 days of hypoglycemia, the Km for 3-O-methyl-D-glucose transport in isolated rat islets, normally 18-20 mM, was 2.5 mM. This provides functional evidence of a profound reduction of high Km glucose transporter in beta cells. In contrast, GLUT-2 was only slightly reduced by hypoglycemia in liver. To determine the effect of prolonged hyperglycemia, we also infused animals with 50% (wt/vol) glucose for 5 days (glucose at 200 +/- 50 mg/dl). Hyperglycemic clamping increased GLUT-2 mRNA by 46% (P = 0.001) whereas proinsulin mRNA doubled (P = 0.001). We conclude that GLUT-2 expression in beta cells, but not liver, is subject to regulation by certain perturbations in blood glucose homeostasis.
据推测,β细胞的葡萄糖转运蛋白(GLUT-2)可能在葡萄糖刺激的胰岛素分泌中起重要作用。为了确定这种转运蛋白是组成性表达还是受调控,我们对清醒不受约束的Wistar大鼠进行葡萄糖稳态干扰,并通过原位杂交对β细胞GLUT-2 mRNA进行定量。低血糖3小时(血糖为29±5mg/dl)后,GLUT-2和胰岛素原mRNA信号密度降至对照大鼠水平的25%。4天后(血糖为57±7mg/dl,而生理盐水灌注的对照大鼠为120±10mg/dl),GLUT-2和胰岛素原mRNA密度分别降低了85%和65%(P = 0.001)。12天后(血糖为54±8mg/dl),GLUT-2 mRNA信号密度无法检测到,而胰岛素原mRNA降低了51%。低血糖12天后,分离的大鼠胰岛中3-O-甲基-D-葡萄糖转运的Km值(正常为18 - 20mM)为2.5mM。这提供了β细胞中高Km葡萄糖转运蛋白显著减少的功能证据。相比之下,低血糖仅使肝脏中的GLUT-2略有减少。为了确定长期高血糖的影响,我们还给动物输注50%(重量/体积)葡萄糖5天(血糖为200±50mg/dl)。高血糖钳夹使GLUT-2 mRNA增加了46%(P = 0.001),而胰岛素原mRNA增加了一倍(P = 0.001)。我们得出结论,β细胞而非肝脏中的GLUT-2表达受血糖稳态某些干扰的调控。