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Modulation of matrix metalloprotease 13 (collagenase 3) gene expression in equine chondrocytes by interleukin 1 and corticosteroids.
- Source :
-
American journal of veterinary research [Am J Vet Res] 1996 Nov; Vol. 57 (11), pp. 1631-4. - Publication Year :
- 1996
-
Abstract
- 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.<br />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.<br />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.<br />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.
- Subjects :
- Animals
Base Sequence
Blotting, Northern
Cartilage, Articular drug effects
Cartilage, Articular enzymology
Collagenases biosynthesis
Gene Expression Regulation, Enzymologic drug effects
Matrix Metalloproteinase 13
Methylprednisolone pharmacology
Methylprednisolone Acetate
Molecular Sequence Data
Cartilage, Articular cytology
Collagenases genetics
Dexamethasone pharmacology
Glucocorticoids pharmacology
Horses
Interleukin-1 pharmacology
Methylprednisolone analogs & derivatives
Subjects
Details
- Language :
- English
- ISSN :
- 0002-9645
- Volume :
- 57
- Issue :
- 11
- Database :
- MEDLINE
- Journal :
- American journal of veterinary research
- Publication Type :
- Academic Journal
- Accession number :
- 8915443