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Updated: Oct 13, 2025
Detecting Migration and Infiltration of Neutrophils in Mice
Published on: February 6, 2020
Ouabain inhibits p38 activation in mice neutrophils
Luiz Henrique Agra Cavalcante-Silva1, Deyse Cristina Madruga Carvalho1, Éssia de Almeida Lima1
1Immunobiotechnology Laboratory, Biotechnology Center, Federal University of Paraíba, João Pessoa, 58051‑900, Brazil.
Abstract:
Ouabain is a cardiac steroid hormone with immunomodulatory effects. It inhibits neutrophils migration induced by different stimuli, but little is known about the mechanisms involved in this effect. Thus, the aim of this study was to evaluate the ouabain effect on chemotactic signaling pathways in neutrophils. For that, mice neutrophils were isolated from bone marrow, treated with ouabain (1, 10, and 100 nM) for 2 h, submitted to transwell chemotaxis assay and flow cytometry analysis of Akt, ERK, JNK, and p38 phosphorylation induced by zymosan. Ouabain treatment (1, 10 and, 100 nM) reduces neutrophil chemotaxis induced by chemotactic peptide fMLP, but this substance did not inhibit Akt, ERK, and JNK activation induced by zymosan. However, ouabain (1 and 10 nM) reduced p38 phosphorylation in zymosan-stimulated neutrophils. These results suggest that ouabain may interfere in neutrophil migration through p38 MAPK inhibition.
Insights
Ouabain, a cardiac steroid hormone, reduces neutrophil migration by inhibiting p38 mitogen-activated protein kinase (MAPK) signaling. This study explores the mechanisms behind ouabain's immunomodulatory effects on neutrophils.
Area of Science:
- Immunology
- Pharmacology
- Cell Biology
Background:
- Ouabain is a cardiac steroid hormone known for its immunomodulatory properties.
- Ouabain inhibits neutrophil migration, but the underlying mechanisms are not fully understood.
Purpose of the Study:
- To investigate the effect of ouabain on neutrophil chemotactic signaling pathways.
- To elucidate the role of specific signaling molecules in ouabain-mediated neutrophil migration inhibition.
Main Methods:
- Isolation of mouse neutrophils from bone marrow.
- Treatment with varying concentrations of ouabain (1, 10, 100 nM).
- Chemotaxis assays and flow cytometry analysis of Akt, ERK, JNK, and p38 phosphorylation.
Main Results:
- Ouabain (1-100 nM) reduced neutrophil chemotaxis induced by fMLP.
- Ouabain did not inhibit Akt, ERK, or JNK activation in zymosan-stimulated neutrophils.
- Ouabain (1-10 nM) decreased p38 phosphorylation in zymosan-stimulated neutrophils.
Conclusions:
- Ouabain interferes with neutrophil migration potentially through the inhibition of p38 MAPK signaling.
- These findings provide insights into the immunomodulatory mechanisms of ouabain in neutrophils.
