HUMAN RETINAL-PIGMENT EPITHELIAL-CELLS CULTURED IN HYPERGLYCEMIC MEDIA ACCUMULATE INCREASED AMOUNTS OF GLYCOSAMINOGLYCAN PRECURSORS

被引:0
作者
MARANO, CW
SZWERGOLD, BS
KAPPLER, F
BROWN, TR
MATSCHINSKY, FM
机构
[1] UNIV PENN, DIABET RES CTR, PHILADELPHIA, PA 19104 USA
[2] UNIV PENN, DEPT BIOCHEM & BIOPHYS, PHILADELPHIA, PA 19104 USA
[3] FOX CHASE CANC INST, DEPT NMR & MED SPECT, PHILADELPHIA, PA 19111 USA
关键词
GLUCOSE; HUMAN RETINAL PIGMENT EPITHELIUM; GLYCOSAMINOGLYCANS; BASEMENT MEMBRANE; DIABETES;
D O I
暂无
中图分类号
R77 [眼科学];
学科分类号
100212 ;
摘要
Confluent human retinal pigmented epithelial cells were cultured on microcarrier beads in the presence of 5.6 or 26 mmol/l glucose with or without the aldose-reductase inhibitor Sorbinil (200-mu-M) for 2 wk. At the end of the incubation period, perchloric acid extracts were prepared and analyzed by P-31 nuclear magnetic resonance spectroscopy. As assessed by this method, the phosphorylated metabolites of cells incubated with 5.6 or 26 mmol/l glucose differed significantly in the concentrations of a number of uridine diphosphate (UDP)-conjugated monosaccharides, which were elevated two- to threefold in cells incubated in 26 mmol/l glucose over control samples. The affected metabolites were identified (through a series of spiking experiments) to be UDP-N-acetylglucosamine, UDP-N-acetylgalactosamine, and UDP-glucuronic acid. Coincubation of the cells with Sorbinil 200-mu-M in the presence of 26 mmol/l glucose had no effect on this accumulation. Under normal circumstances, these molecules selectively and sequentially are incorporated into the polysaccharide chains of glycosaminoglycans (GAGs), whose presence and distribution in the basement membranes is affected adversely by diabetes mellitus. These data suggest that the availability of the monosaccharide precursor is not the rate-limiting step for GAG synthesis in the presence of pathologic glucose concentrations. Thus, the lost GAG content in the basement membranes of diabetic patients may be caused by changes elsewhere in the biosynthesis and/or catabolism of the polysaccharide-linked protein molecules.
引用
收藏
页码:2619 / 2625
页数:7
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