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      Cultured retinal capillary pericytes die by apoptosis after an abrupt fluctuation from high to low glucose levels: a comparative study with retinal capillary endothelial cells.

      Diabetologia

      1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine, Animals, Apoptosis, Bucladesine, pharmacology, Calcimycin, Capillaries, cytology, drug effects, physiology, Cattle, Cell Survival, Cells, Cultured, Coculture Techniques, DNA, analysis, Endothelium, Vascular, Enzyme Inhibitors, Glucose, metabolism, Isoquinolines, Kinetics, Microscopy, Electron, Muscle, Smooth, Vascular, Niacinamide, Piperazines, Retinal Vessels, Signal Transduction, Sulfonamides, Tetradecanoylphorbol Acetate, Time Factors

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          Abstract

          A number of clinical observations concerning cases of glycemic fluctuation have prompted us to study whether or not a rapid change in blood glucose concentration can aggravate retinal microvascular pathology during the early stage of diabetic retinopathy. We conducted a comparative study of retinal capillary pericytes and endothelial cells in vitro. Both types of cells, either in single culture or in co-culture, were initially incubated in medium with high glucose (20-40 mmol/l), followed by a rapid reduction of glucose to 3.5, 1, or 0.5 mmol/1. This type of reduction of extracellular glucose resulted in depletion of intracellular glucose, occurring much faster in pericytes than in endothelial cells. The abrupt reduction in glucose caused pericyte cell shrinkage and nuclear condensation associated with DNA fragmentation, followed by loss of cell viability. All of these pericyte changes are apoptosis-like characteristics. This apoptotic process was prevented by the addition of cycloheximide, a protein synthesis inhibitor, or by platelet-derived growth factor BB, which is known competent factor for pericyte growth. In analysis of signalling pathways during the abrupt fluctuation of glucose, the occurrence of pericyte apoptosis was an intracellular calcium-dependent, protein kinase C and protein kinase A mediated, and poly (ADP-ribose) synthetase-dependent process. Interestingly, a larger degree of DNA fragmentation was observed with a higher magnitude and a longer duration of pre-existing hyperglycaemia. These results suggest that the magnitude and duration of pre-existing hyperglycaemia prime the apoptotic responsiveness of pericytes. Retinal capillary endothelial cells, after an identical glucose fluctuation treatment did not undergo an apoptotic process.

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          8739913

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