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NAD(P)H oxidase mediates the endothelial barrier dysfunction induced by TNF-alpha
Nancy Gertzberg1, Paul Neumann, Victor Rizzo
1Center for Cardiovascular Science, Albany Medical College, NY 12208, USA.
Summary
Tumor necrosis factor-alpha (TNF) increases lung microvessel permeability via NAD(P)H oxidase activation and superoxide anion (O2-*) generation. Inhibiting these pathways restores normal barrier function.
Area of Science:
- Pulmonary vascular biology
- Cellular signaling
- Oxidative stress
Background:
- Tumor necrosis factor-alpha (TNF) is implicated in inflammatory lung diseases.
- Endothelial barrier dysfunction contributes to pulmonary edema.
Purpose of the Study:
- To investigate the role of NAD(P)H oxidase and superoxide anion (O2-*) in TNF-induced pulmonary microvessel endothelial monolayer (PMEM) permeability.
Main Methods:
- Assessed PMEM permeability using Evans blue-labeled albumin clearance.
- Quantified NAD(P)H oxidase components (p47phox, p22phox) via immunofluorescence and Western blot.
- Measured reactive oxygen species (O2-*) using fluorescent probes.
- Utilized antisense oligonucleotides and superoxide dismutase for intervention.
Main Results:
- TNF treatment increased p47phox translocation, p22phox levels, O2-* generation, and PMEM permeability.
- p22phox antisense oligonucleotide blocked TNF-induced changes.
- Superoxide dismutase inhibited TNF-induced O2-* generation and permeability.
Conclusions:
- NAD(P)H oxidase activation and subsequent O2-* generation mediate TNF-induced barrier dysfunction in PMEM.
- Targeting this pathway may offer therapeutic potential for inflammatory lung conditions.