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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; p less than 0.001). There was no change in myoinositol levels over the 24 h period. Sorbitol accumulation occurred at glucose concentration greater than 18.1 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.

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