title: Stress-induced signaling pathways in hyalin chondrocytes: inhibition by Avocado-Soybean Unsaponifiables (ASU).
abstract: OBJECTIVE
  Avocado-Soybean Unsaponifiables (ASU) represent one of the most commonly used drugs for symptomatic osteoarthritis (OA). The mechanisms of its activities are still poorly understood. We investigate here the effects of ASU on signaling pathways in mouse or human chondrocytes.
  
  
  METHODS
  Mouse or human chondrocytes stimulated with interleukin-1beta (IL1beta, 10 ng/ml) and cartilage submitted to a compressive mechanical stress (MS) were studied in the presence or absence of ASU (10 microg/ml). Nuclear factor kappaB (NF-kappaB) activation was assessed by immunoblot, using an I-kappa B alpha antibody, nuclear translocation of NF-kappaB using p65 antibody, and extra-cellular signal-regulated kinase (ERK)1/2 activation using phospho and ERK1/2 antibodies. The binding of the p50/p65 complex on DNA was studied by electrophoretic mobility shift assay.
  
  
  RESULTS
  ASU decrease matrix metalloproteinases-3 and -13 expressions and Prostaglandin E(2) (PGE(2)) release in our model. The degradation of I-kappa B alpha is prevented in the presence of ASU as shown by the persistent expression of I-kappa B alpha protein in the cytosol when chondrocytes are stimulated by IL1beta or MS. Nuclear translocation of the NF-kappaB complex is shown by the decrease of the p65 protein from the cytosol, whereas p65 appears in the nucleus under IL1beta stimulation. This translocation is abolished in the presence of ASU. Moreover, bandshift experiments show an inhibition of the IL1beta-induced binding of p50/p65 complexes to NF-kappaB responsive elements in response to ASU. Finally, among the different mitogen-activated protein kinases known to be induced by IL1beta, ERK1/2 was the sole kinase inhibited by ASU.
  
  
  CONCLUSIONS
  These results demonstrate that ASU express a unique range of activities, which could counteract deleterious processes involved in OA, such as inflammation.
authors: UMR 7079 University Pierre and Marie Curie Paris VI/CNRS, Physiology and Pathophysiology Laboratory, 7 Quai St Bernard, Paris, 75252 Cedex 5, France.
authors: O
authors: O
authors: Gabay
authors: M
authors: M
authors: Gosset
authors: A
authors: A
authors: Levy
authors: C
authors: C
authors: Salvat
authors: C
authors: C
authors: Sanchez
authors: A
authors: A
authors: Pigenet
authors: A
authors: A
authors: Sautet
authors: C
authors: C
authors: Jacques
authors: F
authors: F
authors: Berenbaum
mesh_terms: D053674:Aggrecans / Q000037:antagonists & inhibitors; D000818:Animals; D015153:Blotting, Western / Q000379:methods; D002358:Cartilage, Articular / Q000473:pathology; D018929:Cell Culture Techniques; D019902:Chondrocytes; D024043:Collagen Type II / Q000037:antagonists & inhibitors; D015232:Dinoprostone / Q000378:metabolism; D006801:Humans; D051457:Hyaline Cartilage; D007122:Immunoelectrophoresis / Q000379:methods; D053583:Interleukin-1beta / Q000502:physiology; D020935:MAP Kinase Signaling System / Q000187:drug effects; D051379:Mice; D016328:NF-kappa B / Q000502:physiology*; D010003:Osteoarthritis / Q000188:drug therapy* / Q000473:pathology; D027421:Persea; D010936:Plant Extracts / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects*; D013025:Glycine max; D013314:Stress, Mechanical
chemical_list: D053674:Aggrecans; D024043:Collagen Type II; D053583:Interleukin-1beta; D016328:NF-kappa B; D010936:Plant Extracts; D015232:Dinoprostone