Nicotinamide adenine dinucleotide phosphate oxidase in experimental liver fibrosis: GKT137831 as a novel potential therapeutic agent

Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX) generates reactive oxygen species (ROS) in hepatic stellate cells (HSCs) during liver fibrosis. In response to fibrogenic agonists, such as angiotensin II (Ang II), the NOX1 components form an active complex, including Ras‐related botulinum toxin substrate 1 (Rac1). Superoxide dismutase 1 (SOD1) interacts with the NOX‐Rac1 complex to stimulate NOX activity. NOX4 is also induced in activated HSCs/myofibroblast by increased gene expression. Here, we investigate the role of an enhanced activity SOD1 G37R mutation (SODmu) and the effects of GKT137831, a dual NOX1/4 inhibitor, on HSCs and liver fibrosis. To induce liver fibrosis, wild‐type (WT) and SOD1mu mice were treated with CCl4 or bile duct ligation (BDL). Then, to address the role of NOX‐SOD1‐mediated ROS production in HSC activation and liver fibrosis, mice were treated with a NOX1/4 inhibitor. Fibrosis and ROS generation was assessed by histology and measurement of thiobarbituric acid reactive substances and NOX‐related genes. Primary cultured HSCs isolated from WT, SODmu, and NOX1 knockout (KO) mice were assessed for ROS production, Rac1 activity, and NOX gene expression. Liver fibrosis was increased in SOD1mu mice, and ROS production and Rac1 activity were increased in SOD1mu HSCs. The NOX1/4 inhibitor, GKT137831, attenuated liver fibrosis and ROS production in both SOD1mu and WT mice as well as messenger RNA expression of fibrotic and NOX genes. Treatment with GKT137831 suppressed ROS production and NOX and fibrotic gene expression, but not Rac1 activity, in SOD1mut and WT HSCs. Both Ang II and tumor growth factor beta up‐regulated NOX4, but Ang II required NOX1. Conclusions: SOD1mu induces excessive NOX1 activation through Rac1 in HSCs, causing enhanced NOX4 up‐regulation, ROS generation, and liver fibrosis. Treatment targeting NOX1/4 may be a new therapy for liver fibrosis. (HEPATOLOGY 2012)

[1]  R. Brandes,et al.  Liver fibrosis and hepatocyte apoptosis are attenuated by GKT137831, a novel NOX4/NOX1 inhibitor in vivo. , 2012, Free radical biology & medicine.

[2]  Natalie J Torok,et al.  NOX1/nicotinamide adenine dinucleotide phosphate, reduced form (NADPH) oxidase promotes proliferation of stellate cells and aggravates liver fibrosis induced by bile duct ligation , 2011, Hepatology.

[3]  Y. Gorin,et al.  Myofibroblast differentiation during fibrosis: role of NAD(P)H oxidases. , 2011, Kidney international.

[4]  K. Iwaisako,et al.  The nicotinamide adenine dinucleotide phosphate oxidase (NOX) homologues NOX1 and NOX2/gp91phox mediate hepatic fibrosis in mice , 2011, Hepatology.

[5]  S. Friedman,et al.  Scraping fibrosis: Expressway to the core of fibrosis , 2011, Nature Medicine.

[6]  I. Jacobson,et al.  Managing patients with hepatitis-B-related or hepatitis-C-related decompensated cirrhosis , 2011, Nature Reviews Gastroenterology &Hepatology.

[7]  L. Fuentes-Broto,et al.  Monitoring systemic oxidative stress in an animal model of amyotrophic lateral sclerosis , 2011, Journal of Neurology.

[8]  R. Touyz,et al.  Critical role of Nox4-based NADPH oxidase in glucose-induced oxidative stress in the kidney: implications in type 2 diabetic nephropathy. , 2010, American journal of physiology. Renal physiology.

[9]  Sophie Houngninou-Molango,et al.  First in class, potent, and orally bioavailable NADPH oxidase isoform 4 (Nox4) inhibitors for the treatment of idiopathic pulmonary fibrosis. , 2010, Journal of medicinal chemistry.

[10]  D. Brenner,et al.  CX3CL1‐CX3CR1 interaction prevents carbon tetrachloride‐induced liver inflammation and fibrosis in mice , 2010, Hepatology.

[11]  R. Schwabe,et al.  Role and cellular source of nicotinamide adenine dinucleotide phosphate oxidase in hepatic fibrosis , 2010, Hepatology.

[12]  B. Crestani,et al.  NOX4/NADPH oxidase expression is increased in pulmonary fibroblasts from patients with idiopathic pulmonary fibrosis and mediates TGFβ1-induced fibroblast differentiation into myofibroblasts , 2010, Thorax.

[13]  L. V. van Grunsven,et al.  Advanced glycation end products induce production of reactive oxygen species via the activation of NADPH oxidase in murine hepatic stellate cells. , 2010, Journal of hepatology.

[14]  H. Abboud,et al.  NAD(P)H oxidase mediates TGF-beta1-induced activation of kidney myofibroblasts. , 2010, Journal of the American Society of Nephrology : JASN.

[15]  K. Griendling,et al.  Nox proteins in signal transduction. , 2009, Free radical biology & medicine.

[16]  D. Schuppan,et al.  Targeting liver fibrosis: Strategies for development and validation of antifibrotic therapies , 2009, Hepatology.

[17]  D. Brenner,et al.  Effects of losartan on hepatic expression of nonphagocytic NADPH oxidase and fibrogenic genes in patients with chronic hepatitis C. , 2009, American journal of physiology. Gastrointestinal and liver physiology.

[18]  F. Martinez,et al.  NADPH Oxidase-4 Mediates Myofibroblast Activation and Fibrogenic Responses to Lung Injury , 2009, Nature Medicine.

[19]  K. Ikejima,et al.  Pioglitazone promotes survival and prevents hepatic regeneration failure after partial hepatectomy in obese and diabetic KK‐Ay mice , 2009, Hepatology.

[20]  S. Reese,et al.  Nox‐2 Is a Modulator of Fibrogenesis in Kidney Allografts , 2008, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[21]  F. Gage,et al.  Non-cell-autonomous effect of human SOD1 G37R astrocytes on motor neurons derived from human embryonic stem cells. , 2008, Cell stem cell.

[22]  R. Schwabe,et al.  Reduced nicotinamide adenine dinucleotide phosphate oxidase mediates fibrotic and inflammatory effects of leptin on hepatic stellate cells , 2008, Hepatology.

[23]  B. He,et al.  RXR agonists inhibit high-glucose-induced oxidative stress by repressing PKC activity in human endothelial cells. , 2008, Free radical biology & medicine.

[24]  H. Paulson,et al.  SOD1 mutations disrupt redox-sensitive Rac regulation of NADPH oxidase in a familial ALS model. , 2008, The Journal of clinical investigation.

[25]  D. Cleveland,et al.  Revisiting oxidative damage in ALS: microglia, Nox, and mutant SOD1. , 2008, The Journal of clinical investigation.

[26]  S. Nakano,et al.  Phosphorylated Smad2/3 immunoreactivity in sporadic and familial amyotrophic lateral sclerosis and its mouse model , 2008, Acta Neuropathologica.

[27]  L. Terada,et al.  HIV-1 Tat Activates Dual Nox Pathways Leading to Independent Activation of ERK and JNK MAP Kinases* , 2007, Journal of Biological Chemistry.

[28]  K. Krause,et al.  NOX4 activity is determined by mRNA levels and reveals a unique pattern of ROS generation. , 2007, The Biochemical journal.

[29]  D. Brenner,et al.  NOX in liver fibrosis. , 2007, Archives of biochemistry and biophysics.

[30]  R. Schwabe,et al.  Gene expression profiles during hepatic stellate cell activation in culture and in vivo. , 2007, Gastroenterology.

[31]  D. Brenner,et al.  Gastric bypass surgery improves metabolic and hepatic abnormalities associated with nonalcoholic fatty liver disease. , 2006, Gastroenterology.

[32]  Natalie J Torok,et al.  Phagocytosis of apoptotic bodies by hepatic stellate cells induces NADPH oxidase and is associated with liver fibrosis in vivo , 2006, Hepatology.

[33]  K. Krause,et al.  Decreased blood pressure in NOX1‐deficient mice , 2006, FEBS letters.

[34]  D. Sorescu,et al.  NAD(P)H Oxidase 4 Mediates Transforming Growth Factor-β1–Induced Differentiation of Cardiac Fibroblasts Into Myofibroblasts , 2005, Circulation research.

[35]  Junitsu Ito,et al.  NAD(P)H oxidase plays a crucial role in PDGF‐induced proliferation of hepatic stellate cells , 2005, Hepatology.

[36]  D. Brenner,et al.  Erratum: Liver fibrosis (Journal of Clinical Investigation (2005) 115 (209-218) DOI:10.1172/JCI200524282) , 2005 .

[37]  D. Brenner,et al.  A dual reporter gene transgenic mouse demonstrates heterogeneity in hepatic fibrogenic cell populations , 2004, Hepatology.

[38]  R. Schwabe,et al.  NADPH oxidase signal transduces angiotensin II in hepatic stellate cells and is critical in hepatic fibrosis. , 2003, The Journal of clinical investigation.

[39]  H. Abboud,et al.  Nox4 mediates angiotensin II-induced activation of Akt/protein kinase B in mesangial cells. , 2003, American journal of physiology. Renal physiology.

[40]  P. Bioulac-Sage,et al.  [Mechanisms of hepatic fibrogenesis]. , 2002, Archives de pediatrie : organe officiel de la Societe francaise de pediatrie.

[41]  A. Jesaitis,et al.  Phosphatidic Acid and Diacylglycerol Directly Activate NADPH Oxidase by Interacting with Enzyme Components* , 2001, The Journal of Biological Chemistry.

[42]  R. Schwabe,et al.  Gene delivery of Cu/Zn‐superoxide dismutase improves graft function after transplantation of fatty livers in the rat , 2000, Hepatology.

[43]  D. Cleveland.  From Charcot to SOD1 Mechanisms of Selective Motor Neuron Death in ALS , 1999, Neuron.

[44]  D. Borchelt,et al.  An adverse property of a familial ALS-linked SOD1 mutation causes motor neuron disease characterized by vacuolar degeneration of mitochondria , 1995, Neuron.

[45]  D. C. Carter,et al.  Atomic structure and chemistry of human serum albumin , 1993, Nature.

[46]  J. Haines,et al.  Mutations in Cu/Zn superoxide dismutase gene are associated with familial amyotrophic lateral sclerosis , 1993, Nature.

[47]  S. Friedman,et al.  Activation of cultured rat hepatic lipocytes by Kupffer cell conditioned medium. Direct enhancement of matrix synthesis and stimulation of cell proliferation via induction of platelet-derived growth factor receptors. , 1989, The Journal of clinical investigation.