Moutzouris Dimitrios-Anestis D, Kitsiou Paraskevi V, Talamagas Argiris A, Drossopoulou Garyfallia I, Kassimatis Theodore I, Katsilambros Nikolaos K
First Department of Propaedeutic Medicine, University of Athens Medical School, Laiko University Hospital, Athens, Greece.
Ren Fail. 2007;29(3):353-8. doi: 10.1080/08860220601184126.
GLUTs are specific membrane proteins that transport glucose down a concentration gradient. There have been few studies on their expression in the kidney. The aim of this study was to identify the expression of GLUTs 1, 3, and 4 in HGEC and their regulation under diabetic milieu.
An immortalized cell line of HGEC was used. Cells were cultured in medium containing 5 or 25 mM D-glucose. Western blotting and flow cytometry were used to examine the presence of GLUTs (1, 3, 4) and alterations in expression.
Western blotting analysis revealed that GLUT-1 levels were increased by 53% in HGEC cultured under experimental diabetes compared to cells grown in 5mM glucose. GLUT-3 levels were also increased by 15% under diabetic conditions. GLUT-4 levels were decreased by 20% in diabetes. Fluorescence Activated Cell Sorting (FACS) analysis demonstrated that cell surface expression of GLUT-1 was increased by 28% in cells grown in 25mM glucose. High glucose concentration did not affect cell surface expression of GLUT-3 and GLUT-4.
These findings suggest that depressed GLUT4 expression in glomerulus and overexpression of GLUT-1 and in a lesser extent of GLUT-3 may alter the glucose uptake in these cells. It has been suggested that the overexpression of GLUT-1 in glomerulus, being the major isoform, may lead to the initial pathologic hallmarks of diabetic nephropathy.
葡萄糖转运蛋白(GLUTs)是一类特定的膜蛋白,可顺着浓度梯度转运葡萄糖。关于它们在肾脏中的表达研究较少。本研究旨在确定人肾小球内皮细胞(HGEC)中GLUT1、GLUT3和GLUT4的表达情况以及它们在糖尿病环境下的调控机制。
使用HGEC的永生化细胞系。细胞在含有5或25 mM D-葡萄糖的培养基中培养。采用蛋白质免疫印迹法和流式细胞术检测GLUTs(1、3、4)的存在情况及表达变化。
蛋白质免疫印迹分析显示,与在5 mM葡萄糖中生长的细胞相比,实验性糖尿病条件下培养的HGEC中GLUT-1水平增加了53%。糖尿病条件下GLUT-3水平也增加了15%。糖尿病状态下GLUT-4水平降低了20%。荧光激活细胞分选(FACS)分析表明,在25 mM葡萄糖中生长的细胞中GLUT-1的细胞表面表达增加了28%。高糖浓度不影响GLUT-3和GLUT-4的细胞表面表达。
这些发现表明,肾小球中GLUT4表达降低以及GLUT-1的过表达和程度较轻的GLUT-3过表达可能会改变这些细胞对葡萄糖的摄取。有人提出,作为主要异构体的肾小球中GLUT-1过表达可能导致糖尿病肾病的初始病理特征。