Modulation of matrix metalloprotease 13 (collagenase 3) gene expression in equine chondrocytes by interleukin 1 and corticosteroids

Am J Vet Res. 1996 Nov;57(11):1631-4.


Objective: To determine whether matrix metalloprotease 13 (MMP-13; collagenase 3) is produced by equine chondrocytes and to investigate modulation of its expression by recombinant human interleukin 1 beta (rhIL-1 beta) and corticosteroids.

Procedure: Equine chondrocytes in monolayer culture were stimulated with rhIL-1 beta. Total RNA was extracted, purified, and reverse transcribed into DNA. Using appropriate primers, a putative MMP-13 fragment was amplified by polymerase chain reaction, and cloned into a bacterial vector. The resultant fragment was purified and sequenced, then was used to prepare a digoxigenin-labeled cRNA probe. Monolayer cultures of first-passage chondrocytes were treated with rhIL-1 beta in the presence or absence of dexamethasone (10(-6)M) or methylprednisolone acetate (10(-9)M to 10(-5)M), in addition to positive and negative controls. Cellular RNA was extracted and resolved on agarose gels and subjected to northern blot analysis, using the equine MMP-13 probe.

Results: Reverse transcriptase-polymerase chain reaction enabled isolation of a 0.6-kb fragment of equine MMP-13 cDNA that had 93% homology with the human MMP-13 cDNA sequence. rhIL-1 significantly stimulated MMP-13 expression in the chondrocytes. Methylprednisolone acetate inhibited the stimulatory effects of rhIL-1 in dose-dependent manner that was statistically significant at 10(-5)M.

Conclusions: Novel information was gained on the existence of MMP-13 and its expression in equine chondrocytes, which suggests a possible role for this enzyme in matrix degradation in horses with arthritis.

MeSH terms

  • Animals
  • Base Sequence
  • Blotting, Northern
  • Cartilage, Articular / cytology*
  • Cartilage, Articular / drug effects
  • Cartilage, Articular / enzymology
  • Collagenases / biosynthesis
  • Collagenases / genetics*
  • Dexamethasone / pharmacology*
  • Gene Expression Regulation, Enzymologic / drug effects
  • Glucocorticoids / pharmacology*
  • Horses*
  • Interleukin-1 / pharmacology*
  • Matrix Metalloproteinase 13
  • Methylprednisolone / analogs & derivatives*
  • Methylprednisolone / pharmacology
  • Methylprednisolone Acetate
  • Molecular Sequence Data


  • Glucocorticoids
  • Interleukin-1
  • Methylprednisolone Acetate
  • Dexamethasone
  • Collagenases
  • MMP13 protein, human
  • Matrix Metalloproteinase 13
  • Methylprednisolone