ABSTRACT
BACKGROUND: Persistence of myofibroblasts is believed to contribute to the development of fibrosis in idiopathic pulmonary fibrosis (IPF). Transforming growth factor beta1 (TGFbeta1) irreversibly converts fibroblasts into pathological myofibroblasts, which express smooth muscle alpha-actin (alpha-SMA) and produce extracellular matrix proteins, such as procollagen I (alpha1). Reactive oxygen species produced by NADPH oxidases (NOXs) have been shown to regulate cell differentiation. It was hypothesised that NOX could be expressed in parenchymal pulmonary fibroblasts and could mediate TGFbeta1-stimulated conversion of fibroblasts into myofibroblasts. METHODS: Fibroblasts were cultured from the lung of nine controls and eight patients with IPF. NOX4, alpha-SMA and procollagen I (alpha1) mRNA and protein expression, reactive oxygen species production and Smad2/3 phosphorylation were quantified, in the absence and in the presence of incubation with TGFbeta1. Migration of platelet-derived growth factor (PDGF)-induced fibroblasts was also assessed. RESULTS: It was found that (1) NOX4 mRNA and protein expression was upregulated in pulmonary fibroblasts from patients with IPF and correlated with mRNA expression of alpha-SMA and procollagen I (alpha1) mRNA; (2) TGFbeta1 upregulated NOX4, alpha-SMA and procollagen I (alpha1) expression in control and IPF fibroblasts; (3) the change in alpha-SMA and procollagen I (alpha1) expression in response to TGFbeta1 was inhibited by antioxidants and by a NOX4 small interfering RNA (siRNA); (4) NOX4 modulated alpha-SMA and procollagen I (alpha1) expression by controlling activation of Smad2/3; and (5) NOX4 modulated PDGF-induced fibroblast migration. CONCLUSION: NOX4 is critical for modulation of the pulmonary myofibroblast phenotype in IPF, probably by modulating the response to TGFbeta1 and PDGF.
Subject(s)
Fibroblasts/enzymology , Idiopathic Pulmonary Fibrosis/pathology , Lung/pathology , NADPH Oxidases/biosynthesis , Transforming Growth Factor beta1/pharmacology , Adult , Aged , Cell Differentiation/drug effects , Cell Differentiation/physiology , Cells, Cultured , Female , Fibroblasts/drug effects , Fibroblasts/pathology , Gene Expression Regulation, Enzymologic , Humans , Idiopathic Pulmonary Fibrosis/enzymology , Lung/enzymology , Male , Middle Aged , NADPH Oxidase 4 , NADPH Oxidases/genetics , NADPH Oxidases/physiology , Platelet-Derived Growth Factor/pharmacology , RNA, Messenger/genetics , Reactive Oxygen Species/metabolism , Reverse Transcriptase Polymerase Chain Reaction/methods , Smad2 Protein/metabolism , Smad3 Protein/metabolism , Up-RegulationABSTRACT
BACKGROUND: Viscum album (VA) extracts are widely used in cancer therapy and are known to be cytotoxic to tumors and endothelial cells. Angiogenesis plays an important role in the growth, sustenance and metastasis of tumors. Inhibition of angiogenesis is now being explored as a new therapeutic avenue for cancer. MATERIALS AND METHODS: The cytotoxicity of VA extracts was analyzed by Annexin V labeling and propidium iodide uptake in EA-hy926 endothelial cells. The antiangiogenic effect was studied in vitro by treating the EA-hy926 cells in matrigel and subsequent analysis of vascular formation. Computer-assisted image analysis of vascular formation was analyzed to quantify the in vitro data. In vivo studies were performed by implanting matrigel +/- VA extracts in Balb/C mice that had been subjected to IP treatment with VA extracts. RESULTS: The combination of systemic and intra- matrigel treatment with the VA Qu Spez extract caused significant inhibition of angiogenesis. The VA P extract treatment showed insignificant change in vessel formation. CONCLUSION: These results may provide novel guidelines towards improved strategies using VA extracts based on the inhibition of angiogenesis of tumors.
Subject(s)
Angiogenesis Inhibitors/pharmacology , Apoptosis/drug effects , Endothelium, Vascular/drug effects , Neovascularization, Physiologic/drug effects , Plant Extracts/pharmacology , Viscum album/chemistry , Animals , Collagen/metabolism , Drug Combinations , Endothelium, Vascular/pathology , Female , Image Processing, Computer-Assisted , Laminin/metabolism , Mice , Mice, Inbred BALB C , Proteoglycans/metabolismABSTRACT
Intravenous immunoglobulin (IVIG) is increasingly used in the treatment of diverse immune-mediated disorders. Since several preparations of IVIG are available for therapy, it is possible that different manufacturing processes might influence clinical efficacy of IVIG. An insight into the mechanisms of action of such different IVIG preparations is therefore necessary that will provide further guidelines for the utility of IVIG preparations in autoimmune and inflammatory diseases. Since endothelial cells (EC) influence the inflammatory process via production of cytokines, chemokines and expression of adhesive molecules, we analyzed the anti-inflammatory effect on EC of two IVIG preparations: caprylated IVIG (IVIG-C) versus solvent/detergent-treated IVIG (IVIG-SD) preparation. We found that both IVIG preparations inhibit in an equivalent manner, the expression of different pro-inflammatory factors such as IL-6, IL-8, GM-CSF, IL-1beta and TNF-alpha and the adhesion molecules ICAM-1 and VCAM-1. Our results thus suggest that the caprylate while inactivating the virus and enhancing the yield of IgG during IVIG formulation, does not modulate the immunomodulatory properties of IVIG at EC level and that the two preparations show similar anti-inflammatory effects.
Subject(s)
Immunoglobulins, Intravenous/chemistry , Immunoglobulins, Intravenous/pharmacology , Immunologic Factors/pharmacology , Caprylates , Cell Culture Techniques/methods , Cytokines/metabolism , Endothelial Cells/immunology , Endothelial Cells/metabolism , Humans , Inflammation/immunology , Inflammation/prevention & control , Intercellular Adhesion Molecule-1/metabolism , Umbilical Veins/immunology , Vascular Cell Adhesion Molecule-1/metabolismABSTRACT
Viscum album (VA) preparations (Iscador) consist of aqueous extracts from different types of European mistletoe. Biologically active components of VA extracts include mistletoe lectins (ML) and viscotoxins. The treatment with VA extracts or with purified ML has been shown to be associated with tumor regression in several in vivo experimental models of tumoral implantation. The mechanisms underlying the anti-tumoral activity of VA or ML are complex and involve apoptosis, angiogenesis and immunomodulation. This review provides an account of the current status of the understanding of the VA-associated immunomodulation in various cell types including lymphoblastoid, monocytic or endothelial cell lines.
Subject(s)
Immunologic Factors , Plant Extracts/pharmacology , Plant Proteins/pharmacology , Angiogenesis Inhibitors/pharmacology , Apoptosis/drug effects , B-Lymphocytes/drug effects , Humans , Monocytes/drug effects , T-Lymphocytes/drug effectsABSTRACT
Intravenous immunoglobulin (IVIg) has been used in the treatment of primary and secondary antibody deficiencies for over 25 years. It is a safe preparation with no long-term side effects. IVIg was first demonstrated to be effective in autoimmune disorders, two decades ago, in the treatment of acute immune thrombocytopenia. Since then, the therapeutic efficacy of IVIg has been established in Guillain Barré syndrome (GBS), chronic inflammatory demyelinating polyneuropathy (CIDP), myasthenia gravis (MG), dermatomyositis (DM), Kawasaki syndrome and the prevention of graft-versus-host disease in recipients of allogeneic bone marrow transplants and reported in a large number of other autoimmune and systemic inflammatory conditions.
