Effect of matrix depleting agents on the expression of chondrocyte metabolism by equine chondrocytes.
This study was carried out to investigate the effect of two enzymes (collagenase and chondroitinase) and two cytokines/metabolites (interleukin-1beta and retinoic acid) of known catabolic activity on the expression of cartilage metabolism/phenotype in equine articular cartilage. Articular cartilage explants from 11 horses (5-13 years old) were treated for 48 h and assayed for total sulphated glycosaminoglycan (GAG), the incorporation of 35S-sulphate, collagen degradation and mRNA expression of the proteoglycans collagen II, collagen IIA, collagen III, collagen IX, collagen X, collagen XI and glyceraldehyde-3-phosphate (GAPDH). Purified collagenase and retinoic acid were responsible for increased GAG loss from the tissues. Chondroitinase, responsible for catalysing the elimination of glucuronate residues from chondroitin A, B and C (Chondroitinase ABC) and retinoic acid treatment induced an inhibition of proteoglycan synthesis, whereas collagenase treatment did not. Collagenase activity was correlated with increased appearance of the CB11B epitope and type II collagen denaturation. By RT-PCR there was evidence of expression of altered collagen type IIA in purified collagenase treated tissues.
Iqbal
J
J
Veterinary Basic Sciences Department, Royal Veterinary College, Hawkshead Lane, North Mymms, Hatfield, Hertfordshire AL9 7TA, UK.
Bird
J L
JL
Hollander
A P
AP
Bayliss
M T
MT
D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology; D019902:Chondrocytes / Q000187:drug effects / Q000378:metabolism*; D019765:Chondroitin ABC Lyase / Q000494:pharmacology*; D003094:Collagen / Q000235:genetics / Q000378:metabolism*; D017364:Collagenases / Q000494:pharmacology*; D005986:Glyceraldehyde 3-Phosphate / Q000235:genetics / Q000378:metabolism; D006025:Glycosaminoglycans / Q000378:metabolism*; D006736:Horses; D007375:Interleukin-1 / Q000494:pharmacology*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D014212:Tretinoin / Q000494:pharmacology*
D006025:Glycosaminoglycans; D007375:Interleukin-1; D012333:RNA, Messenger; D005986:Glyceraldehyde 3-Phosphate; D014212:Tretinoin; D003094:Collagen; D017364:Collagenases; D019765:Chondroitin ABC Lyase