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Poly (ADP) ribose synthetase inhibition in alveolar macrophages undergoing hypoxia and reoxygenation
Anton S McCourtie1, Alexander S Farivar, Steven M Woolley
1Department of Surgery, Division of Cardiothoracic Surgery, University of Washington Medical Center, Seattle, WA 98195, USA.
Background:
Inhibition of the nuclear enzyme poly ribose synthetase (PARS) protects against in vivo lung ischemia reperfusion injury (LIRI). The effectiveness of intratracheal treatment suggests that PARS inhibition may primarily modulate alveolar macrophage (AM) activation. These studies attempted to characterize the effects of PARS on AM activation in response to oxidative stress.
Methods:
Primary cultures of AM were rendered hypoxic for 2 h and reoxygenated for up to 4 h. Cells were preincubated with INO-1001, a specific PARS inhibitor 1 h prior to hypoxia. Gel shift assays characterized nuclear factor kappa B (NFkappaB), and enzyme linked immunosorbent assay quantitated chemokine/cytokine protein secretion.
Results:
Hypoxia and reoxygenation resulted in an increase in the early nuclear translocation of NFkappaB, and an increase in the secretion of the cytokine tumor necrosis factor-alpha (TNF-alpha), chemokines macrophage inflammatory protein (MIP-1alpha), monocyte chemoattractant protein one (MCP-1) and cytokine induced neutrophil chemoattractant (CINC). Pretreatment of AM with INO-1001 decreased both the early translocation of NFkappaB and the production of TNF-alpha (p<0.05) and MIP-1alpha p=0.02, but did not affect CINC or MCP-1 production.
Conclusions:
These findings indicate that PARS inhibition in the AM blunts their response to oxidative stress and may help explain the protective effects of intratracheal PARS inhibition in LIRI.
Insights
Poly ribose synthetase (PARS) inhibition protects against lung injury by reducing alveolar macrophage activation. PARS inhibition blunts the inflammatory response to oxidative stress in macrophages.
Area of Science:
- Immunology
- Cellular Biology
- Pulmonary Medicine
Background:
- Poly ribose synthetase (PARS) inhibition shows protective effects against lung ischemia reperfusion injury (LIRI).
- Intratracheal administration suggests PARS inhibition may target alveolar macrophage (AM) activation.
- Studies investigated PARS's role in AM activation during oxidative stress.
Purpose of the Study:
- To characterize the effects of PARS inhibition on AM activation.
- To investigate the impact of PARS on oxidative stress-induced inflammatory responses in AMs.
- To explore the mechanism underlying PARS inhibition's protective role in LIRI.
Main Methods:
- Primary AM cultures were subjected to hypoxia and reoxygenation.
- Cells were pretreated with INO-1001, a PARS inhibitor.
- NF-kappaB activation was assessed using gel shift assays, and cytokine/chemokine secretion was measured by ELISA.
Main Results:
- Hypoxia-reoxygenation increased NF-kappaB translocation and secretion of TNF-alpha, MIP-1alpha, MCP-1, and CINC.
- INO-1001 pretreatment significantly reduced NF-kappaB translocation and TNF-alpha and MIP-1alpha production.
- PARS inhibition did not affect CINC or MCP-1 production.
Conclusions:
- PARS inhibition in AMs attenuates their response to oxidative stress.
- This blunting of inflammatory response may explain the protective effects of PARS inhibition in LIRI.
- Targeting PARS in AMs presents a potential therapeutic strategy for LIRI.
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