The effect of high glucose on polyol pathway activity and myoinositol metabolism in cultured human endothelial cells

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Abstract
Cultured human endothelial cells were incubated with high glucose (27.5 mmol/l) supplemented medium for 0, 2, 6, 12, 24 and 48 h. Lactate dehydrogenase was assayed as a measure of cell number. Sorbitol and myoinositol were determined as trimethyl silyl derivatives using gas chromatography mass spectrometry. Sorbitol, after an initial lag phase of 2 h, accumulated rapidly to plateau at 2 to 3 fold control value by 24 h, no further increase being seen over a further 24 h (Baseline, 1.31±0.36, 24 h 3.50±0.74 pmol sorbitol/ unit lactate dehydrogenase; p18.1 mmol/l and only returned to baseline levels after 24 h of incubation in normal medium (glucose levels 5.9 mmol/l). Sorbitol levels are increased by incubation of endothelial cells at high glucose levels but myoinositol levels remain unchanged.