During pathological conditions, oxidative burst generates hyaluronan (HA) fragmentation with consequent increase in small HA oligosaccharide levels. These fragments are able to stimulate inflammatory response, in different cell types, by activating the CD44 and both the toll-like receptors 4 (TLR-4) and 2 (TLR-2). CD44 and TLRs stimulation in turn activates the NF-kB that induces the production of several pro-inflammatory mediators that amplify and perpetuate inflammation. We aimed to study the effect of the antioxidant SOD mimic synthetic manganese porphyrin, Mn(III) 5,10,15,20-tetrakis(N-methylpyridinium-2-yl)porphyrin (MnTM-2-PyP5+) in preventing HA degradation in mouse articular chondrocytes stimulated with iron (II) plus ascorbate. Fe (II) plus ascorbate stimulation induced oxidative burst confirmed by high levels of hydroxyl radical and peroxynitrite production, and increase in both lipid peroxidation and HA degradation. HA fragments produced high mRNA expression and the related protein production of CD44, TLR-4 and TLR-2, NF-kB activation and a significant up-regulation of the inflammatory cytokines, tumor necrosis factor alpha (TNF-alpha), interleukin-1beta (IL-1beta), and other pro-inflammatory mediators, such as matrix metalloprotease 13 (MMP-13) and inducible nitric oxide synthase (iNOS). Treatment of cells with MnTM-2-PyP(5+) was able to attenuate the oxidative burst, HA degradation and NF-kB activation, markedly decreased CD44, and TLRs mRNA expression and the related protein synthesis, as well as the up-regulated inflammatory mediators production. Addition to cells of HA blocking peptide PEP-1 (HABP) significantly reduced all the inflammatory parameters up-regulated by Fe (II) plus ascorbate, and strengthens MnTM-2-PyP(5+) activity. These data suggest that HA degradation plays a key role in the earliest inflammatory response of cartilage and antioxidants could be a useful tool to prevent the amplification of the mechanism.

SOD MIMIC MnTM-2-PyP(5+) REDUCES INFLAMMATION INDUCED BY DEGRADED HYALURONAN IN CHONDROCYTES STIMULATED WITH Fe(II) PLUS ASCORBATE

D'ASCOLA, ANGELA;AVENOSO, Angela;Scuruchi M.;MICALI, Antonio Girolamo;PUZZOLO, Domenico;CALATRONI, Alberto;CAMPO, Salvatore Giuseppe;CAMPO, Giuseppe Maurizio
2012-01-01

Abstract

During pathological conditions, oxidative burst generates hyaluronan (HA) fragmentation with consequent increase in small HA oligosaccharide levels. These fragments are able to stimulate inflammatory response, in different cell types, by activating the CD44 and both the toll-like receptors 4 (TLR-4) and 2 (TLR-2). CD44 and TLRs stimulation in turn activates the NF-kB that induces the production of several pro-inflammatory mediators that amplify and perpetuate inflammation. We aimed to study the effect of the antioxidant SOD mimic synthetic manganese porphyrin, Mn(III) 5,10,15,20-tetrakis(N-methylpyridinium-2-yl)porphyrin (MnTM-2-PyP5+) in preventing HA degradation in mouse articular chondrocytes stimulated with iron (II) plus ascorbate. Fe (II) plus ascorbate stimulation induced oxidative burst confirmed by high levels of hydroxyl radical and peroxynitrite production, and increase in both lipid peroxidation and HA degradation. HA fragments produced high mRNA expression and the related protein production of CD44, TLR-4 and TLR-2, NF-kB activation and a significant up-regulation of the inflammatory cytokines, tumor necrosis factor alpha (TNF-alpha), interleukin-1beta (IL-1beta), and other pro-inflammatory mediators, such as matrix metalloprotease 13 (MMP-13) and inducible nitric oxide synthase (iNOS). Treatment of cells with MnTM-2-PyP(5+) was able to attenuate the oxidative burst, HA degradation and NF-kB activation, markedly decreased CD44, and TLRs mRNA expression and the related protein synthesis, as well as the up-regulated inflammatory mediators production. Addition to cells of HA blocking peptide PEP-1 (HABP) significantly reduced all the inflammatory parameters up-regulated by Fe (II) plus ascorbate, and strengthens MnTM-2-PyP(5+) activity. These data suggest that HA degradation plays a key role in the earliest inflammatory response of cartilage and antioxidants could be a useful tool to prevent the amplification of the mechanism.
2012
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11570/2469021
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