Genetic deficiency of NADPH oxidase does not diminish, but rather enhances, LPS-induced acute inflammatory responses in vivo

Linus Pauling Institute, Oregon State University, Corvallis, OR 97331, USA
Free Radical Biology and Medicine (Impact Factor: 5.74). 03/2009; 46(6):791-798. DOI: 10.1016/j.freeradbiomed.2008.12.003


Reactive oxygen species (ROS) and oxidative stress are thought to play a central role in the etiology of cell dysfunction and tissue damage in sepsis. However, there is limited and controversial evidence from in vivo studies that ROS mediate cell signaling processes that elicit acute inflammatory responses during sepsis. Because NADPH oxidase is one of the main cellular sources of ROS, we investigated the role of this enzyme in lipopolysaccharide (LPS)-induced acute inflammation in vivo, utilizing mice deficient in the gp91phox or p47phox subunits of NADPH oxidase. Age-and body weight-matched C57BL/6J wild-type (WT) and gp91phox−/− and p47phox−/− mice were injected ip with 50 μg LPS or saline vehicle and sacrificed at various time points up to 24 h. We found that LPS-induced acute inflammatory responses in serum and tissues were not significantly diminished in gp91phox−/− and p47phox−/− mice compared to WT mice. Rather, genetic deficiency of NADPH oxidase was associated with enhanced gene expression of inflammatory mediators and increased neutrophil recruitment to lung and heart. Furthermore, no protection from LPS-induced septic death was observed in either knockout strain. Our findings suggest that NADPH oxidase-mediated ROS production and cellular redox signaling do not promote, but instead limit, LPS-induced acute inflammatory responses in vivo.


Available from: Hao Wei, Mar 20, 2015
  • Source
    • "Reactive oxygen species (ROS) play a vital role in LPStriggered macrophage activation by regulating intracellular reduction-oxidation (redox) sensitive signaling pathways and nuclear transcription factors, such as nuclear factor-í µí¼…B (NFí µí¼…B) and nuclear factor-erythroid 2 p45-related factor 2 (Nrf2)123. Overproduction of ROS in macrophages leads to excessive expression of cytokines and inflammatory factors, resulting in atheromatous plaques, acute inflammation, tissue injury, and sepsis [4]. However, most cell types have developed defensive mechanisms to counteract ROS genera- tion [5, 6] . "
    [Show abstract] [Hide abstract] ABSTRACT: Reactive oxygen species (ROS) and oxidative stress are thought to play a central role in potentiating macrophage activation, causing excessive inflammation, tissue damage, and sepsis. Recently, we have shown that punicalagin (PUN) exhibits anti-inflammatory activity in LPS-stimulated macrophages. However, the potential antioxidant effects of PUN in macrophages remain unclear. Revealing these effects will help understand the mechanism underlying its ability to inhibit excessive macrophage activation. Hemeoxygenase-1 (HO-1) exhibits antioxidant activity in macrophages. Therefore, we hypothesized that HO-1 is a potential target of PUN and tried to reveal its antioxidant mechanism. Here, PUN treatment increased HO-1 expression together with its upstream mediator nuclear factor-erythroid 2 p45-related factor 2 (Nrf2). However, specific inhibition of Nrf2 by brusatol (a specific Nrf2 inhibitor) dramatically blocked PUN-induced HO-1 expression. Previous research has demonstrated that the PI3K/Akt pathway plays a critical role in modulating Nrf2/HO-1 protein expression as an upstream signaling molecule. Here, LY294002, a specific PI3K/Akt inhibitor, suppressed PUN-induced HO-1 expression and led to ROS accumulation in macrophages. Furthermore, PUN inhibited LPS-induced oxidative stress in macrophages by reducing ROS and NO generation and increasing superoxide dismutase (SOD) 1 mRNA expression. These findings provide new perspectives for novel therapeutic approaches using antioxidant medicines and compounds against oxidative stress and excessive inflammatory diseases including tissue damage, sepsis, and endotoxemic shock.
    Full-text · Article · Apr 2015 · Mediators of Inflammation
  • Source
    • "In addition to oxidation, ROS have immunomodulatory activity that can improve the immune response to an infection. In fact, ROS are considered second messengers in immune system signaling and act upstream of diverse biological responses involving transcriptional regulation, cytokines expression, and inflammation [28]. In this study, LPS-induced NF-κB activation, as determined by Western blot analysis, and ROS production were significantly inhibited with pretreatment of clinically relevant concentrations of propofol. "
    [Show abstract] [Hide abstract] ABSTRACT: During an infection, lipopolysaccharide (LPS) stimulates the production of reactive oxygen species (ROS), which is mediated, in large part, by nicotinamide adenine dinucleotide phosphate (NADPH) oxidases (NOXs); NOX2 is the major NOX isoform found in the macrophage cell membrane. While the immunomodulatory activity of propofol is highly documented, its effect on the LPS-induced NOX2/ROS/NF-κB signaling pathway in macrophages has not been addressed. In present study, we used murine macrophage cell line RAW264.7 pretreated with propofol and stimulated with LPS. IL-6 and TNF-α expression, ROS production, and NOX activity were determined. Results showed that propofol attenuated LPS-induced TNF-α and IL-6 expression. Moreover, LPS-stimulated phosphorylation of NF-κB and generation of ROS were weakened in response to propofol. Propofol also reduced LPS-induced NOX activity and expression of gp91phox and p47phox. We conclude that propofol modulates LPS signaling in macrophages by reducing NOX-mediated production of TNF-α and IL-6.
    Full-text · Article · Nov 2013 · Clinical and Developmental Immunology
  • Source
    • "NOX is recognized as a critical mediator of inflammatory responses in nonimmune systems, such as neuroinflammation, gastric inflammation, colon inflammation, lung inflammation, and vascular inflammation. An anti-inflammatory activity of NOX enzymes, also mentioned in NOX2- and p47phox-dificient mice, found that LPS-induced systemic inflammatory responses were significantly enhanced by genetic deficiency of NOX [55]. However, the precise molecular mechanisms accountable for the installation of NADPH oxidase-mediated inflammation are still an unsettled issue. "
    [Show abstract] [Hide abstract] ABSTRACT: Proteinuria is an independent risk factor for end-stage renal disease (ESRD) (Shankland, 2006). Recent studies highlighted the mechanisms of podocyte injury and implications for potential treatment strategies in proteinuric kidney diseases (Zhang et al., 2012). Reactive oxygen species (ROS) are cellular signals which are closely associated with the development and progression of glomerular sclerosis. NADPH oxidase is a district enzymatic source of cellular ROS production and prominently expressed in podocytes (Zhang et al., 2010). In the last decade, it has become evident that NADPH oxidase-derived ROS overproduction is a key trigger of podocyte injury, such as renin-angiotensin-aldosterone system activation (Whaley-Connell et al., 2006), epithelial-to-mesenchymal transition (Zhang et al., 2011), and inflammatory priming (Abais et al., 2013). This review focuses on the mechanism of NADPH oxidase-mediated ROS in podocyte injury under different pathophysiological conditions. In addition, we also reviewed the therapeutic perspectives of NADPH oxidase in kidney diseases related to podocyte injury.
    Full-text · Article · Nov 2013
Show more