High glucose level inhibits capacitative Ca2+ influx in cultured rat mesangial cells by a protein kinase C-dependent mechanism

Diabetologia
P MenèF Pugliese

Abstract

In cultured mesangial cells (MC), capacitative Ca2+ influx via store-operated channels (SOC) is potentiated by agents that release Ca2+ from intracellular stores, and inhibited by protein kinase C (PKC). Cells grown under high glucose conditions, as a model of the diabetic microenvironment, display reduced Ca2+ signalling in response to vasoconstrictors, probably due to downregulation by elevated PKC activity. Since SOC might be relevant to this phenomenon, we assessed Ca2+ influx by microfluorometry of fura-2-loaded rat MC cultured for 5 days in normal (5.5 mmol/l, NG) or high glucose (30 mmol/l, HG). The addition of 1-10 mmol/l Ca2+ to NG cells equilibrated in Ca(2+)-free media induced an immediate Ca2+ influx with a free cytosolic Ca2+ ([Ca2+]i) plateau of 155 +/- 50 and 318 +/- 114 nmol/l, respectively. Basal influx was reduced to 88 +/- 8 and 145 +/- 17 nmol/l [Ca2+]i (1-10 mmol/l Ca2+, p < 0.01) by 30 mmol/l D-glucose. This effect of HG was confirmed by Mn2+ quenching of fura-2, indicating reduced entry of divalent cations via the capacitative pathway. Equimolar L-glucose had no effect on Ca2+ influx, consistent with a non-osmotic mechanism. Arginine vasopressin (10 mumol/l) elicited weaker release of stored Ca2+ and subs...Continue Reading

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