High glucose-induced apoptosis through store-operated calcium entry and calcineurin in human umbilical vein endothelial cells.
Diabetes mellitus causes multiple cardiovascular complications. Previous studies have shown that prolonged exposure (96 h) of human umbilical vein endothelial cells (HUVECs) to hyperglycemia causes a significant increase in apoptosis. We report here that this increase in apoptosis is associated with an increase in Ca(2+) current (whole cell patch-clamp recorded) resulting from Ca(2+) entry mediated by store-operated channels (SOCs). The number of apoptotic cells after prolonged high glucose (HG, 30 mmol/L) exposure was significantly reduced in the presence of the SOC inhibitor 2-APB or of La(3+). A marked increase (approximately 80%) in Ca(2+)-dependent calcineurin (CN-A) phosphatase activity also occurred after prolonged HG exposure. Prolonged HG exposure-induced increase in CN-A activity was prevented by 2-APB, and selective CN-A phosphatase inhibition by FK506 or calmodulin inhibition by calmidazolium decreased HG-induced apoptosis. Blocking hydrogen peroxide production using catalase or inhibiting the tyrosine kinase pp60(src) during prolonged exposure to HG, resulted in a marked decrease in apoptosis and was further associated with a significant reduction in CN-A phosphatase activity. The results demonstrate a significant role for Ca(2+) entry in HG-induced apoptosis in HUVECs, and suggest that this role is mediated via H(2)O(2) generation and the action of the Ca(2+)-activated protein phosphatase calcineurin.
Tamareille
Sophie
S
Centre National de la Recherche Scientifique, Unité Mixte de Recherche 8078, Université Paris XI, Hôpital Marie Lannelongue, 92350 Le Plessis Robinson, France.
Mignen
Olivier
O
Capiod
Thierry
T
Rücker-Martin
Catherine
C
Feuvray
Danielle
D
D017209:Apoptosis; D001896:Boron Compounds / Q000494:pharmacology; D019703:Calcineurin / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000187:drug effects / Q000378:metabolism; D002147:Calmodulin / Q000037:antagonists & inhibitors; D002374:Catalase / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000037:antagonists & inhibitors / Q000378:metabolism; D006943:Hyperglycemia / Q000378:metabolism / Q000503:physiopathology; D007093:Imidazoles / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000037:antagonists & inhibitors; D016559:Tacrolimus / Q000494:pharmacology; D014471:Umbilical Veins / Q000166:cytology
D001896:Boron Compounds; D015220:Calcium Channels; D002147:Calmodulin; D007093:Imidazoles; C031938:calmidazolium; D006861:Hydrogen Peroxide; C109986:2-aminoethoxydiphenyl borate; D002374:Catalase; D016392:Proto-Oncogene Proteins pp60(c-src); C120817:calcineurin phosphatase; D019703:Calcineurin; D010744:Phosphoric Monoester Hydrolases; D005947:Glucose; D002118:Calcium; D016559:Tacrolimus