Reactive oxygen species-mediated p38 MAPK regulates carbon nanotube-induced fibrogenic and angiogenic responses

Abstract Single-walled carbon nanotubes (SWCNTs) are fibrous nanoparticles that are being used widely for various applications including drug delivery. SWCNTs are currently under special attention for possible cytotoxicity. Recent reports suggest that exposure to nanoparticles leads to pulmonary fibrosis. We report that SWCNT-mediated interplay of fibrogenic and angiogenic regulators leads to increased angiogenesis, which is a novel finding that furthers the understanding of SWCNT-induced cytotoxicity. SWCNTs induce fibrogenesis through reactive oxygen species-regulated phosphorylation of p38 mitogen-activated protein kinase (MAPK). Activation of p38 MAPK by SWCNTs led to the induction of transforming growth factor (TGF)-β1 as well as vascular endothelial growth factor (VEGF). Both TGF-β1 and VEGF contributed significantly to the fibroproliferative and collagen-inducing effects of SWCNTs. Interestingly, a positive feedback loop was observed between TGF-β1 and VEGF. This interplay of fibrogenic and angiogenic mediators led to increased angiogenesis in response to SWCNTs. Overall this study reveals key signalling molecules involved in SWCNT-induced fibrogenesis and angiogenesis.

[1]  R. Homer,et al.  Semaphorin 7A plays a critical role in TGF-β1–induced pulmonary fibrosis , 2007, The Journal of experimental medicine.

[2]  R. Chapela,et al.  Concentration, biosynthesis and degradation of collagen in idiopathic pulmonary fibrosis. , 1986, Thorax.

[3]  P. Baron,et al.  Unusual inflammatory and fibrogenic pulmonary responses to single-walled carbon nanotubes in mice. , 2005, American journal of physiology. Lung cellular and molecular physiology.

[4]  A. Nicholson,et al.  Interstitial vascularity in fibrosing alveolitis. , 2003, American journal of respiratory and critical care medicine.

[5]  M. Turner-Warwick,et al.  Precapillary Systemic-pulmonary Anastomoses , 1963, Thorax.

[6]  L. Gold,et al.  Regulation of the effects of TGF-beta 1 by activation of latent TGF-beta 1 and differential expression of TGF-beta receptors (T beta R-I and T beta R-II) in idiopathic pulmonary fibrosis. , 2001, Thorax.

[7]  J. Kern,et al.  Role of Smad2/3 and p38 MAP kinase in TGF‐β1‐induced epithelial–mesenchymal transition of pulmonary epithelial cells , 2011, Journal of cellular physiology.

[8]  M. Burdick,et al.  ENA-78 is an important angiogenic factor in idiopathic pulmonary fibrosis. , 2001, American journal of respiratory and critical care medicine.

[9]  S. Spivack,et al.  Regulation of antioxidant enzymes in lung after oxidant injury. , 1994, Environmental health perspectives.

[10]  E. Crouch,et al.  Pathobiology of pulmonary fibrosis. , 1990, The American journal of physiology.

[11]  Malgorzata Zakrzewska,et al.  Phosphorylation of Fibroblast Growth Factor (FGF) Receptor 1 at Ser777 by p38 Mitogen-Activated Protein Kinase Regulates Translocation of Exogenous FGF1 to the Cytosol and Nucleus , 2008, Molecular and Cellular Biology.

[12]  Dihua Yu,et al.  Pathway Enhances Endothelial Cell Migration 1-activated p 38 Signaling β Cancer Cells by the Heregulin-Up-Regulation of Vascular Endothelial Growth Factor in Breast , 2001 .

[13]  P. Carmeliet.  Mechanisms of angiogenesis and arteriogenesis , 2000, Nature Medicine.

[14]  A. Mui,et al.  Release of biologically active TGF-beta1 by alveolar epithelial cells results in pulmonary fibrosis. , 2003, American journal of physiology. Lung cellular and molecular physiology.

[15]  G. Maurer,et al.  Oncostatin M-enhanced vascular endothelial growth factor expression in human vascular smooth muscle cells involves PI3K-, p38 MAPK-, Erk1/2- and STAT1/STAT3-dependent pathways and is attenuated by interferon-γ , 2011, Basic Research in Cardiology.

[16]  Bing-Hua Jiang,et al.  Phosphatidylinositol-3-kinase/akt regulates bleomycin-induced fibroblast proliferation and collagen production. , 2010, American journal of respiratory cell and molecular biology.

[17]  W. Fiers,et al.  The p38/RK mitogen‐activated protein kinase pathway regulates interleukin‐6 synthesis response to tumor necrosis factor. , 1996, The EMBO journal.

[18]  C. Daniels,et al.  Imatinib mesylate inhibits the profibrogenic activity of TGF-beta and prevents bleomycin-mediated lung fibrosis. , 2004, The Journal of clinical investigation.

[19]  Constance E. Brinckerhoff,et al.  Matrix metalloproteinases: a tail of a frog that became a prince , 2002, Nature Reviews Molecular Cell Biology.

[20]  M. Sporn,et al.  Transforming growth factor type beta: rapid induction of fibrosis and angiogenesis in vivo and stimulation of collagen formation in vitro. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[21]  M. Burdick,et al.  Neutralization of the CXC chemokine, macrophage inflammatory protein-2, attenuates bleomycin-induced pulmonary fibrosis. , 1999, Journal of immunology.

[22]  P. Vincent,et al.  p38 MAPK activation by TGF-beta1 increases MLC phosphorylation and endothelial monolayer permeability. , 2002, American journal of physiology. Lung cellular and molecular physiology.

[23]  V. Kagan,et al.  The role of nanotoxicology in realizing the ‘helping without harm’ paradigm of nanomedicine: lessons from studies of pulmonary effects of single‐walled carbon nanotubes , 2010, Journal of internal medicine.

[24]  R. Whyte,et al.  The CXC chemokines, IL-8 and IP-10, regulate angiogenic activity in idiopathic pulmonary fibrosis. , 1997, Journal of immunology.

[25]  R. Jain,et al.  C-X-C receptor type 4 promotes metastasis by activating p38 mitogen-activated protein kinase in myeloid differentiation antigen (Gr-1)-positive cells , 2010, Proceedings of the National Academy of Sciences.

[26]  M. Selman,et al.  MMP-1: the elder of the family. , 2005, The international journal of biochemistry & cell biology.

[27]  R. Ji,et al.  Activation of p38 Mitogen-activated Protein Kinase in Spinal Microglia Contributes to Incision-induced Mechanical Allodynia , 2009, Anesthesiology.

[28]  M. Burdick,et al.  IFN-gamma-inducible protein-10 attenuates bleomycin-induced pulmonary fibrosis via inhibition of angiogenesis. , 1999, Journal of immunology.

[29]  K. Meyer,et al.  Vascular endothelial growth factor in bronchoalveolar lavage from normal subjects and patients with diffuse parenchymal lung disease. , 2000, The Journal of laboratory and clinical medicine.

[30]  J. James,et al.  Pulmonary toxicity of single-wall carbon nanotubes in mice 7 and 90 days after intratracheal instillation. , 2003, Toxicological sciences : an official journal of the Society of Toxicology.

[31]  P. Baron,et al.  Inhalation vs. aspiration of single-walled carbon nanotubes in C57BL/6 mice: inflammation, fibrosis, oxidative stress, and mutagenesis. , 2008, American journal of physiology. Lung cellular and molecular physiology.

[32]  K. Irie,et al.  Purification and Identification of a Major Activator for p38 from Osmotically Shocked Cells , 1996, The Journal of Biological Chemistry.

[33]  A. Pardo,et al.  Idiopathic Pulmonary Fibrosis: Prevailing and Evolving Hypotheses about Its Pathogenesis and Implications for Therapy , 2001, Annals of Internal Medicine.

[34]  Jiahuai Han,et al.  Pro-inflammatory Cytokines and Environmental Stress Cause p38 Mitogen-activated Protein Kinase Activation by Dual Phosphorylation on Tyrosine and Threonine (*) , 1995, The Journal of Biological Chemistry.

[35]  S. Barni,et al.  In situ assessment of oxidant and nitrogenic stress in bleomycin pulmonary fibrosis , 2006, Histochemistry and Cell Biology.

[36]  F. Martinez,et al.  Mechanisms of pulmonary fibrosis. , 2004, Annual review of medicine.

[37]  M. Gaestel,et al.  CREB is activated by UVC through a p38/HOG‐1‐dependent protein kinase , 1997, The EMBO journal.

[38]  A. Vollmar,et al.  Caffeic acid phenethyl ester inhibits PDGF-induced proliferation of vascular smooth muscle cells via activation of p38 MAPK, HIF-1α, and heme oxygenase-1. , 2011, Journal of natural products.

[39]  R. Weichselbaum,et al.  Activation of p38 Mitogen-activated Protein Kinase by c-Abl-dependent and -independent Mechanisms* , 1996, The Journal of Biological Chemistry.

[40]  M. Müller,et al.  Differential immunolocalization of VEGF in rat and human adult lung, and in experimental rat lung fibrosis: Light, fluorescence, and electron microscopy , 1999, The Anatomical record.

[41]  L Bibbs,et al.  A MAP kinase targeted by endotoxin and hyperosmolarity in mammalian cells. , 1994, Science.

[42]  T. Webb,et al.  Comparative pulmonary toxicity assessment of single-wall carbon nanotubes in rats. , 2003, Toxicological sciences : an official journal of the Society of Toxicology.

[43]  H. Moses,et al.  Transforming growth factor beta 1-induced changes in cell migration, proliferation, and angiogenesis in the chicken chorioallantoic membrane , 1990, The Journal of cell biology.

[44]  R. Clark,et al.  Fibronectin mediates adherence of rat alveolar type II epithelial cells via the fibroblastic cell-attachment domain. , 1986, The Journal of clinical investigation.

[45]  G. Oberdörster,et al.  Nanotoxicology: An Emerging Discipline Evolving from Studies of Ultrafine Particles , 2005, Environmental health perspectives.

[46]  K. Strasser-Weippl,et al.  Increased angiogenesis in chronic idiopathic myelofibrosis: vascular endothelial growth factor as a prominent angiogenic factor. , 2007, Human pathology.

[47]  K. Brown,et al.  Pigment epithelium-derived factor in idiopathic pulmonary fibrosis: a role in aberrant angiogenesis. , 2004, American journal of respiratory and critical care medicine.

[48]  M. Peão,et al.  Neoformation of blood vessels in association with rat lung fibrosis induced by bleomycin , 1994, The Anatomical record.

[49]  Vincent Castranova,et al.  Dispersion of single-walled carbon nanotubes by a natural lung surfactant for pulmonary in vitro and in vivo toxicity studies , 2010, Particle and Fibre Toxicology.

[50]  B. Hogan,et al.  Expression of transforming growth factor beta 2 RNA during murine embryogenesis. , 1989, Development.

[51]  E. Panzhinskiy,et al.  Mitogen-activated protein kinase phosphatase-1 is a key regulator of hypoxia-induced vascular endothelial growth factor expression and vessel density in lung. , 2011, The American journal of pathology.

[52]  L. Gold,et al.  Regulation of the effects of TGF-β1 by activation of latent TGF-β1 and differential expression of TGF-β receptors (TβR-I and TβR-II) in idiopathic pulmonary fibrosis , 2001 .

[53]  G. Karakiulakis,et al.  Hypoxia modulates the effects of transforming growth factor-beta isoforms on matrix-formation by primary human lung fibroblasts. , 2003, Cytokine.

[54]  J. Huh,et al.  Formononetin accelerates wound repair by the regulation of early growth response factor-1 transcription factor through the phosphorylation of the ERK and p38 MAPK pathways. , 2011, International immunopharmacology.

[55]  R. McAnulty,et al.  Pathogenesis of lung fibrosis and potential new therapeutic strategies. , 1995, Experimental nephrology.

[56]  Liying Wang,et al.  Direct Fibrogenic Effects of Dispersed Single-Walled Carbon Nanotubes on Human Lung Fibroblasts , 2010, Journal of toxicology and environmental health. Part A.

[57]  R. D. du Bois,et al.  Transforming growth factors-beta 1, -beta 2, and -beta 3 stimulate fibroblast procollagen production in vitro but are differentially expressed during bleomycin-induced lung fibrosis. , 1997, The American journal of pathology.