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Antiapoptotic effect of ouabain on human umbilical vein endothelial cells
Lucia Trevisi1, Barbara Visentin, Federico Cusinato
1Dipartimento di Farmacologia ed Anestesiologia, Università di Padova, Padua, Italy.
Abstract:
The present study investigates the effect of ouabain on caspase-3 activation in human umbilical vein endothelial cells (HUVEC). Ouabain (EC(50) 20 nM) reduced caspase-3 activity in HUVEC treated for 24h in a medium deprived of fibroblast growth factor (FGF). Incubation for 5h in the absence of both FGF and serum produced an increase in caspase-3 activity that was completely abolished by 100 nM ouabain. Pretreatment with the phosphatidylinositol 3 kinase (PI-3K) inhibitor, wortmannin, prevented the protective effect of ouabain against serum deprivation. Furthermore, Western blotting analysis revealed an increase in phosphorylation of extracellular signal-regulated kinases (ERK-1 and ERK-2) induced by 100nM ouabain in serum-deprived cells. In accord, pretreatment of HUVEC with PD98059, inhibitor of the ERK pathway, abrogated the effect of ouabain. Our results show that ouabain has an antiapoptotic effect on HUVEC through the activation of PI-3K and ERK dependent pathways.
Insights
Ouabain demonstrates an anti-apoptotic effect on human umbilical vein endothelial cells (HUVEC) by reducing caspase-3 activation. This protective action is mediated through the activation of phosphatidylinositol 3-kinase (PI-3K) and extracellular signal-regulated kinase (ERK) pathways.
Area of Science:
- Cardiovascular Biology
- Cell Signaling
- Apoptosis Research
Background:
- Caspase-3 activation is a key mediator of apoptosis.
- Endothelial cell apoptosis contributes to vascular dysfunction.
- Ouabain's role in endothelial cell apoptosis requires further elucidation.
Purpose of the Study:
- To investigate the effect of ouabain on caspase-3 activation in human umbilical vein endothelial cells (HUVEC).
- To elucidate the signaling pathways involved in ouabain's effect on HUVEC apoptosis.
Main Methods:
- Human umbilical vein endothelial cells (HUVEC) were treated with ouabain under various conditions (serum/FGF deprivation).
- Caspase-3 activity was measured.
- Western blotting was used to assess protein phosphorylation (ERK-1/2).
- Specific inhibitors (wortmannin for PI-3K, PD98059 for ERK) were employed.
Main Results:
- Ouabain (20 nM) reduced caspase-3 activity in HUVEC deprived of FGF.
- Ouabain (100 nM) abolished increased caspase-3 activity induced by serum/FGF deprivation.
- The protective effect of ouabain was prevented by the PI-3K inhibitor wortmannin.
- Ouabain increased ERK-1/2 phosphorylation in serum-deprived HUVEC.
- Inhibition of the ERK pathway abrogated ouabain's effect.
Conclusions:
- Ouabain exhibits an anti-apoptotic effect on HUVEC.
- This effect is mediated via the activation of phosphatidylinositol 3-kinase (PI-3K) and extracellular signal-regulated kinase (ERK) dependent pathways.
