Cytokine and chemokine gene expression of IL-1beta stimulated equine articular chondrocytes.
OBJECTIVE
To evaluate mRNA expression of several proinflammatory and anti-inflammatory cytokines and chemokines in equine unstimulated and interleukin-1beta (IL-1beta)-stimulated chondrocytes.


METHODS
In vitro experiment using equine chondrocyte cultures.


METHODS
Whole articular cartilage from metacarpophalangeal joints (n=5 horses; 10 fetlocks).


METHODS
Chondrocyte monolayer cultures were established from digested adult equine articular cartilage and stimulated with 5 ng/mL of recombinant human IL-1beta. RNA was extracted from the cells 24 hours after stimulation. IL-1beta, IL-4, IL-6, IL-8, tumor necrosis factor-alpha (TNF-alpha), and ubiquitin (house keeping gene) mRNA expression were investigated by real-time RT-PCR.


RESULTS
IL-1beta, IL-6, and IL-8 mRNA were expressed in unstimulated chondrocytes from macroscopically normal joints and were significantly up-regulated after stimulation (5/5 horses). IL-4 mRNA was not detected in any samples (0/5 horses). TNF-alpha mRNA, by comparison, was expressed in 2/5 unstimulated samples and in all stimulated samples but a considerable sample variation in response to IL-1beta stimulation was observed.


CONCLUSIONS
Equine chondrocytes express mRNA for several proinflammatory cytokines and chemokines and IL-1beta modulates their expression.


CONCLUSIONS
Chondrocytes express proinflammatory cytokines and chemokines capable of modulating a local inflammatory cascade in articular cartilage, which could potentially lead to focal degradation and osteoarthritis.
David
Florent
F
Département de Sciences Cliniques, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-Hyacinthe, Québec, Canada J2S 7C6.
Farley
Judith
J
Huang
Hong
H
Lavoie
Jean-Pierre
JP
Laverty
Sheila
S
D000818:Animals; D002358:Cartilage, Articular / Q000166:cytology*; D019902:Chondrocytes / Q000187:drug effects / Q000276:immunology*; D016207:Cytokines / Q000096:biosynthesis* / Q000235:genetics; D017931:DNA Primers; D005786:Gene Expression Regulation; D006736:Horses / Q000276:immunology*; D053583:Interleukin-1beta / Q000494:pharmacology; D015850:Interleukin-6 / Q000096:biosynthesis / Q000235:genetics; D016209:Interleukin-8 / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000235:genetics
D016207:Cytokines; D017931:DNA Primers; D053583:Interleukin-1beta; D015850:Interleukin-6; D016209:Interleukin-8; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha