AstraZeneca Diabetes and Obesity Drug Discovery, CVGI iMED, 3S42C Mereside, Alderley Park, Macclesfield, Cheshire SK10 4TG, UK.
J Mol Endocrinol. 2011 Jul 18;47(1):59-67. doi: 10.1530/JME-10-0157. Print 2011 Aug.
Glucokinase (GK) plays a critical role in controlling blood glucose; GK activators have been shown to stimulate insulin secretion acutely both in vitro and in vivo. Sustained stimulation of insulin secretion could potentially lead to β-cell exhaustion; this study examines the effect of chronic GK activation on β-cells. Gene expression and insulin secretion were measured in rodent islets treated in vitro with GKA71 for 72 h. Key β-cell gene expression was measured in rat, mouse and global GK heterozygous knockout mouse islets (gk(del/wt)). Insulin secretion, after chronic exposure to GKA71, was measured in perifused rat islets. GKA71 acutely increased insulin secretion in rat islets in a glucose-dependent manner. Chronic culture of mouse islets with GKA71 in 5 mmol/l glucose significantly increased the expression of insulin, IAPP, GLUT2, PDX1 and PC1 and decreased the expression of C/EBPβ compared with 5 mmol/l glucose alone. Similar increases were shown for insulin, GLUT2, IAPP and PC1 in chronically treated rat islets. Insulin mRNA was also increased in GKA71-treated gk(del/wt) islets. No changes in GK mRNA were observed. Glucose-stimulated insulin secretion was improved in perifused rat islets following chronic treatment with GKA71. This was associated with a greater insulin content and GK protein level. Chronic treatment of rodent islets with GKA71 showed an upregulation of key β-cell genes including insulin and an increase in insulin content and GK protein compared with glucose alone.
葡萄糖激酶(GK)在控制血糖方面起着至关重要的作用;已证明 GK 激活剂在体外和体内均能急性刺激胰岛素分泌。持续刺激胰岛素分泌可能导致β细胞衰竭;本研究检查了慢性 GK 激活对β细胞的影响。在体外用 GKA71 处理啮齿动物胰岛 72 小时后,测量了基因表达和胰岛素分泌。在大鼠、小鼠和全球 GK 杂合子敲除小鼠胰岛(gk(del/wt))中测量了关键β细胞基因表达。在灌注的大鼠胰岛中测量了慢性暴露于 GKA71 后胰岛素的分泌。GKA71 以葡萄糖依赖性方式急性增加大鼠胰岛的胰岛素分泌。在 5 mmol/l 葡萄糖中,用 GKA71 慢性培养的小鼠胰岛显著增加了胰岛素、IAPP、GLUT2、PDX1 和 PC1 的表达,并降低了 C/EBPβ的表达与单独 5 mmol/l 葡萄糖相比。在慢性处理的大鼠胰岛中也显示出胰岛素、GLUT2、IAPP 和 PC1 的相似增加。在 GKA71 处理的 gk(del/wt)胰岛中,胰岛素 mRNA 也增加了。未观察到 GK mRNA 的变化。在慢性 GKA71 处理后,灌注的大鼠胰岛的葡萄糖刺激胰岛素分泌得到改善。这与胰岛素含量和 GK 蛋白水平的增加有关。与单独葡萄糖相比,用 GKA71 慢性处理的啮齿动物胰岛显示关键β细胞基因的上调,包括胰岛素,以及胰岛素含量和 GK 蛋白的增加。