scholarly journals Palytoxin-induced cell death cascade in bovine aortic endothelial cells

2006 ◽  
Vol 291 (4) ◽  
pp. C657-C667 ◽  
Author(s):  
William P. Schilling ◽  
Deborah Snyder ◽  
William G. Sinkins ◽  
Mark Estacion
Keyword(s):  
Endothelial Cells ◽  
Cell Death ◽  
Fluorescent Protein ◽  
Time Lapse ◽  
Cell Lysis ◽  
Monovalent Cation ◽  
Aortic Endothelial Cells ◽  
Bovine Aortic Endothelial Cells ◽  
Cell Death Cascade ◽  
Aortic Endothelial

The plasmalemmal Na+-K+-ATPase (NKA) pump is the receptor for the potent marine toxin palytoxin (PTX). PTX binds to the NKA and converts the pump into a monovalent cation channel that exhibits a slight permeability to Ca2+. However, the ability of PTX to directly increase cytosolic free Ca2+ concentration ([Ca2+]i) via Na+ pump channels and to initiate Ca2+ overload-induced oncotic cell death has not been examined. Thus the purpose of this study was to determine the effect of PTX on [Ca2+]i and the downstream events associated with cell death in bovine aortic endothelial cells. PTX (3–100 nM) produced a graded increase in [Ca2+]i that was dependent on extracellular Ca2+. The increase in [Ca2+]i initiated by 100 nM PTX was blocked by pretreatment with ouabain with an IC50 < 1 μM. The elevation in [Ca2+]i could be reversed by addition of ouabain at various times after PTX, but this required much higher concentrations of ouabain (0.5 mM). These results suggest that the PTX-induced rise in [Ca2+]i occurs via the Na+ pump. Subsequent to the rise in [Ca2+]i, PTX also caused a concentration-dependent increase in uptake of the vital dye ethidium bromide (EB) but not YO-PRO-1. EB uptake was also blocked by ouabain added either before or after PTX. Time-lapse video microscopy showed that PTX ultimately caused cell lysis as indicated by release of transiently expressed green fluorescent protein (molecular mass 27 kDa) and rapid uptake of propidium iodide. Cell lysis was 1) greatly delayed by removing extracellular Ca2+ or by adding ouabain after PTX, 2) blocked by the cytoprotective amino acid glycine, and 3) accompanied by dramatic membrane blebbing. These results demonstrate that PTX initiates a cell death cascade characteristic of Ca2+ overload.

Maitotoxin-induced cell death cascade in bovine aortic endothelial cells: divalent cation specificity and selectivity

2004 ◽  
Vol 287 (2) ◽  
pp. C345-C356 ◽  
Author(s):  
Brian J. Wisnoskey ◽  
Mark Estacion ◽  
William P. Schilling
Keyword(s):  
Endothelial Cells ◽  
Cell Death ◽  
Single Cell ◽  
Binding Proteins ◽  
Single Cell Level ◽  
Cell Level ◽  
Aortic Endothelial Cells ◽  
Bovine Aortic Endothelial Cells ◽  
Membrane Blebs ◽  
Cell Death Cascade

The maitotoxin (MTX)-induced cell death cascade in bovine aortic endothelial cells (BAECs), a model for Ca2+ overload-induced toxicity, reflects three sequential changes in plasmalemmal permeability. MTX initially activates Ca2+-permeable, nonselective cation channels (CaNSC) and causes a massive increase in cytosolic free Ca2+ concentration ([Ca2+]i). This is followed by the opening of large endogenous cytolytic/oncotic pores (COP) that allow molecules <800 Da to enter the cell. The cells then lyse not by rupture of the plasmalemma but through the activation of a “death” channel that lets large proteins (e.g., 140–160 kDa) leave the cell. These changes in permeability are accompanied by the formation of membrane blebs. In this study, we took advantage of the well-known differences in affinity of various Ca2+-binding proteins for Ca2+ and Sr2+ vs. Ba2+ to probe their involvement in each phase of the cell death cascade. Using fluorescence techniques at the cell population level (cuvette-based) and at the single-cell level (time-lapse videomicroscopy), we found that the replacement of Ca2+ with either Sr2+ or Ba2+ delayed both MTX-induced activation of COP, as indicated by the uptake of ethidium bromide, and subsequent cell lysis, as indicated by the uptake of propidium iodide or the release of cell-associated green fluorescent protein. MTX-induced responses were mimicked by ionomycin and were significantly delayed in BAPTA-loaded cells. Experiments at the single-cell level revealed that Ba2+ not only delayed the time to cell lysis but also caused desynchronization of the lytic phase. Last, membrane blebs, which were numerous and spherical in Ca2+-containing solutions, were poorly defined and greatly reduced in number in the presence of Ba2+. Taken together, these results suggest that intracellular high-affinity Ca2+-binding proteins are involved in the MTX-induced changes in plasmalemmal permeability that are responsible for cell demise.

scholarly journals Autocrine Bradykinin Release Promotes Ischemic Preconditioning-Induced Cytoprotection in Bovine Aortic Endothelial Cells

2020 ◽  
Vol 21 (8) ◽  
pp. 2965 ◽  
Author(s):  
Alessandro Bellis ◽  
Daniela Sorriento ◽  
Antonella Fiordelisi ◽  
Raffaele Izzo ◽  
Junichi Sadoshima ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Death ◽  
Ischemic Preconditioning ◽  
Molecular Mechanisms ◽  
Tissue Kallikrein ◽  
Aortic Endothelial Cells ◽  
Akt Activation ◽  
Bovine Aortic Endothelial Cells ◽  
Hoe 140 ◽  
Aortic Endothelial

The aims of this study were to assess whether ischemic preconditioning (PC) induces bradykinin (Bk) synthesis in bovine aortic endothelial cells (bAECs) and, if so, to explore the molecular mechanisms by which this peptide provides cytoprotection against hypoxia. PC was induced by exposing bAECs to three cycles of 15 min of hypoxia followed by 15 min of reoxygenation. Bk synthesis peaked in correspondence to the early and late phases of PC (10−12 M and 10−11 M, respectively) and was abolished by a selective tissue kallikrein inhibitor, aprotinin. Stimulation with exogenous Bk at concentrations of 10−12 M and 10−11 M reduced the cell death induced by 12 h of hypoxia by 50%. Pretreatment with HOE−140, a Bk receptor 2 (BKR2) inhibitor, in bAECs exposed to 12 h of hypoxia, abrogated the cytoprotective effect of early and late PC, whereas des-Arg-HOE-140, a Bk receptor 1 (BKR1) inhibitor, affected only the late PC. In addition, we found that PC evoked endocytosis and the recycling of BKR2 during both the early and late phases, and that inhibition of these pathways affected PC-mediated cytoprotection. Finally, we evaluated the activation of PKA and Akt in the presence or absence of BKR2 inhibitor. HOE-140 abrogated PKA and Akt activation during both early and late PC. Consistently, BKR2 inhibition abolished cross-talk between PKA and Akt in PC. In bAECs, Bk-synthesis evoked by PC mediates the protection against both apoptotic and necrotic hypoxia-induced cell death in an autocrine manner, by both BKR2- and BKR1-dependent mechanisms.

Effects of Dipyrone on Prostaglandin Production by Human Platelets and Cultured Bovine Aortic Endothelial Cells

1983 ◽  
Vol 49 (02) ◽  
pp. 132-137 ◽  
Author(s):  
A Eldor ◽  
G Polliack ◽  
I Vlodavsky ◽  
M Levy
Keyword(s):  
Endothelial Cells ◽  
Arachidonic Acid ◽  
Competitive Inhibition ◽  
Inhibitory Effect ◽  
Human Platelets ◽  
Ionophore A23187 ◽  
Aortic Endothelial Cells ◽  
Bovine Aortic Endothelial Cells ◽  
Aortic Endothelial

SummaryDipyrone and its metabolites 4-methylaminoantipyrine, 4-aminoantipyrine, 4-acetylaminoantipyrine and 4-formylaminoan- tipyrine inhibited the formation of thromboxane A2 (TXA2) during in vitro platelet aggregation induced by ADP, epinephrine, collagen, ionophore A23187 and arachidonic acid. Inhibition occurred after a short incubation (30–40 sec) and depended on the concentration of the drug or its metabolites and the aggregating agents. The minimal inhibitory concentration of dipyrone needed to completely block aggregation varied between individual donors, and related directly to the inherent capacity of their platelets to synthesize TXA2.Incubation of dipyrone with cultured bovine aortic endothelial cells resulted in a time and dose dependent inhibition of the release of prostacyclin (PGI2) into the culture medium. However, inhibition was abolished when the drug was removed from the culture, or when the cells were stimulated to produce PGI2 with either arachidonic acid or ionophore A23187.These results indicate that dipyrone exerts its inhibitory effect on prostaglandins synthesis by platelets or endothelial cells through a competitive inhibition of the cyclooxygenase system.

scholarly journals Ethanolamine metabolism in cultured bovine aortic endothelial cells.

1990 ◽  
Vol 265 (13) ◽  
pp. 7195-7201
Author(s):  
B A Lipton ◽  
E P Davidson ◽  
B H Ginsberg ◽  
M A Yorek
Keyword(s):  
Endothelial Cells ◽  
Aortic Endothelial Cells ◽  
Bovine Aortic Endothelial Cells ◽  
Aortic Endothelial

Microcarrier cultivation of bovine aortic endothelial cells in spinner vessels and a membrane stirred bioreactor

1996 ◽  
Vol 18 (3) ◽  
pp. 193-206 ◽  
Author(s):  
Johannes M�thing ◽  
Sevim Duvar ◽  
Susann Nerger ◽  
Heino B�ntemeyer ◽  
J�rgen Lehmann
Keyword(s):  
Endothelial Cells ◽  
Aortic Endothelial Cells ◽  
Stirred Bioreactor ◽  
Bovine Aortic Endothelial Cells ◽  
Aortic Endothelial
Sign in / Sign up

Export Citation Format

Share Document