Regulation of Ca2+ movements by cyclovirobuxine D in ECV304 endothelial cells.
In Fura-2/loaded ECV304 endothelial cells cyclovirobuxine D promoted a transient increase in cytosolic free Ca2+ originating from both an intracellular pool sensitive to the endoplasmic reticulum Ca2+-ATPase inhibitor thapsigargin and the extracellular space. The intracellular pool was apparently different from that mobilized by other agents acting through IP3 generation. The integrity of the plasma membrane was an absolute requirement. In cells treated with digitonin, cyclovirobuxine D did not promote any Ca2+ release from the intracellular stores even at high concentrations and in the absence or presence of thapsigargin or sodium azide, the inhibitors of the endoplasmic reticular or mitochondrial Ca2+ uptake. Furthermore, cyclovirobuxine D was effective in halting the persistent increase in cytosolic Ca2+ caused by thapsigargin, inhibiting the operation of the "capacitative" Ca2+ membrane channels as demonstrated by the decrease in the extent of both Ca2+-overshoot and Mn2+ influx. Additional effects of cyclovirobuxine D included a depolarization of plasma membrane apparently related to an enhanced influx of Na+ from the extracellular space. The results obtained indicate that cyclovirobuxine D markedly affects intracellular Ca2+ homeostasis in ECV304 endothelial cells by both promoting a discharge of intracellular pools and by interfering with the operation of store-dependent channels via plasma membrane depolarization.
Grossini
Elena
E
Dipartimento di Scienze Mediche, Facoltà di Medicina e Chirurgia, Università del Piemonte Orientale A. Avogadro, via Solaroli 17, I-28100 Novara, Italy. grossini@med.unipmn.it
Avanzi
Giancarlo
G
Gallicchio
Margherita
M
Molinari
Claudio
C
Vacca
Giovanni
G
Bellomo
Giorgio
G
D002118:Calcium / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism / Q000502:physiology; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects; D019284:Thapsigargin / Q000494:pharmacology
D004365:Drugs, Chinese Herbal; D004791:Enzyme Inhibitors; C032939:cyclovirobuxine D; D019284:Thapsigargin; D000252:Calcium-Transporting ATPases; D002118:Calcium