Inhibition of small HA fragment activity and stimulation of A2A adenosine receptor pathway limit apoptosis and reduce cartilage damage in experimental arthritis

Histochem Cell Biol. 2015 May;143(5):531-43. doi: 10.1007/s00418-014-1298-7. Epub 2014 Dec 16.

Abstract

Recent studies have found that the inactivation of small hyaluronan (HA) fragments originating from native HA during inflammation reduced the inflammatory response in models of experimental arthritis. The stimulation of adenosine receptors A2A reduced inflammation by inhibiting NF-kB activation. The combination of both treatments was significantly more effective than either of the individual treatments. The aim of this study was to further investigate the effects of a combined treatment using the HA inhibitor Pep-1 and a selective A2AR agonist (CV-1808) on the structure and ultrastructure of the articular cartilage and on apoptosis in a model of collagen-induced arthritis (CIA) in mice. Arthritic mice were treated with Pep-1 and/or CV-1808 intraperitoneally daily for 20 days. At day 35, the hind limbs were processed for light microscopy (hematoxylin/eosin and Safranin-O-Fast Green) and for transmission and scanning electron microscopy. CIA increased IL-6, caspase-3 and caspase-7 mRNA expression and the related protein levels in arthritic articular cartilage, and significantly increased concentrations of Bcl-2-associated X protein (Bax), while B cell-lymphoma-2 protein (Bcl-2) was markedly reduced. The combined Pep-1/CV-1808 treatment significantly reduced CIA injury, particularly at the highest doses, demonstrated by the presence of Safranin-O-positive cartilage, with a smooth surface and normal chondrocytes in the superficial, intermediate and deep zones. Morphological data and histological scoring were strongly supported by the reduction in inflammation and apoptotic markers. The results further support the role of HA degradation and A2A receptors in arthritis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine / analogs & derivatives*
  • Adenosine / pharmacology
  • Adenosine A2 Receptor Agonists / pharmacology*
  • Animals
  • Anti-Inflammatory Agents / pharmacology*
  • Apoptosis / drug effects*
  • Apoptosis Regulatory Proteins / genetics
  • Apoptosis Regulatory Proteins / metabolism
  • Arthritis, Experimental / chemically induced
  • Arthritis, Experimental / drug therapy*
  • Arthritis, Experimental / metabolism
  • Arthritis, Experimental / pathology
  • Cartilage, Articular / drug effects*
  • Cartilage, Articular / metabolism
  • Cartilage, Articular / ultrastructure
  • Chondrocytes / drug effects
  • Chondrocytes / metabolism
  • Chondrocytes / pathology
  • Collagen Type II
  • Drug Therapy, Combination
  • Freund's Adjuvant
  • Hyaluronic Acid / antagonists & inhibitors*
  • Hyaluronic Acid / metabolism
  • Inflammation Mediators / metabolism
  • Joints / drug effects*
  • Joints / metabolism
  • Joints / ultrastructure
  • Male
  • Mice, Inbred DBA
  • Microscopy, Electron, Scanning
  • Microscopy, Electron, Transmission
  • Peptides / pharmacology*
  • Receptor, Adenosine A2A / drug effects*
  • Receptor, Adenosine A2A / metabolism
  • Severity of Illness Index
  • Signal Transduction / drug effects
  • Time Factors

Substances

  • Adenosine A2 Receptor Agonists
  • Anti-Inflammatory Agents
  • Apoptosis Regulatory Proteins
  • Collagen Type II
  • Inflammation Mediators
  • Peptides
  • Receptor, Adenosine A2A
  • glycyl-alanyl-histidyl-tryptophyl-glutaminyl-phenylalanyl-asparagyl-alanyl-leucyl-threonyl-valyl-arginine
  • Hyaluronic Acid
  • Freund's Adjuvant
  • 2-phenylaminoadenosine
  • Adenosine