TY - JOUR
T1 - Modulation of equine articular chondrocyte messenger RNA levels following brief exposures to recombinant equine interleukin-1β
AU - Takafuji, V. A.
AU - Howard, R. D.
AU - Ward, D. L.
AU - Sharova, L. V.
AU - Crisman, M. V.
PY - 2005/6/15
Y1 - 2005/6/15
N2 - The effect of recombinant equine IL-1β (EqIL-1β) on steady-state mRNA levels of equine articular chondrocytes in high-density monolayer culture was investigated using a customized cDNA array analysis. Total RNA samples isolated from chondrocytes cultured in media alone or with the addition of 1 ng/ml EqIL-1β for 1-, 3-, and 6-h durations of exposure were reverse transcribed, radiolabeled, and hybridized to a customized 380-target cDNA array. Means of duplicate log base 2 transformed hybridization signals were normalized to equine glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mean signal intensities. Differentially expressed transcripts were identified using a two-stage mixed linear analysis of variance model (Statistical Analysis Software, Cary, NC). A time-dependent pattern was observed in the number of transcripts increased ≥two-fold in response to EqIL-1β after 1, 3 and 6 h (1, 2 and 109 transcripts, respectively). At 6 h of EqIL-1β stimulation, signal intensities for 88 cDNA targets with purported function in processes related to cell cycle, intracellular signaling, transcription, translation, extracellular matrix turnover, and inflammation, as well as a number of cDNAs lacking homology to previously reported cDNA sequences, were increased >two-fold and were associated with p < 0.05. Principal component analysis identified a vector component (∼10% of the total variation) corresponding to a potential EqIL-1β co-regulation of cell cycle associated gene transcription. These results support and expand our existing comprehension of the complex role of IL-1 in modulated chondrocyte gene expression and suggest the involvement of specific target gene up-regulation and activation of downstream inflammatory cascade mediators. This study adds to the current understanding of the molecular events associated with an IL-1 induced inflammation and pathobiologic processes that may be associated with the development of equine osteoarthritis.
AB - The effect of recombinant equine IL-1β (EqIL-1β) on steady-state mRNA levels of equine articular chondrocytes in high-density monolayer culture was investigated using a customized cDNA array analysis. Total RNA samples isolated from chondrocytes cultured in media alone or with the addition of 1 ng/ml EqIL-1β for 1-, 3-, and 6-h durations of exposure were reverse transcribed, radiolabeled, and hybridized to a customized 380-target cDNA array. Means of duplicate log base 2 transformed hybridization signals were normalized to equine glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mean signal intensities. Differentially expressed transcripts were identified using a two-stage mixed linear analysis of variance model (Statistical Analysis Software, Cary, NC). A time-dependent pattern was observed in the number of transcripts increased ≥two-fold in response to EqIL-1β after 1, 3 and 6 h (1, 2 and 109 transcripts, respectively). At 6 h of EqIL-1β stimulation, signal intensities for 88 cDNA targets with purported function in processes related to cell cycle, intracellular signaling, transcription, translation, extracellular matrix turnover, and inflammation, as well as a number of cDNAs lacking homology to previously reported cDNA sequences, were increased >two-fold and were associated with p < 0.05. Principal component analysis identified a vector component (∼10% of the total variation) corresponding to a potential EqIL-1β co-regulation of cell cycle associated gene transcription. These results support and expand our existing comprehension of the complex role of IL-1 in modulated chondrocyte gene expression and suggest the involvement of specific target gene up-regulation and activation of downstream inflammatory cascade mediators. This study adds to the current understanding of the molecular events associated with an IL-1 induced inflammation and pathobiologic processes that may be associated with the development of equine osteoarthritis.
KW - Chondrocytes
KW - Gene expression
KW - Interleukin-1β
KW - Osteoarthritis
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UR - http://www.scopus.com/inward/citedby.url?scp=19344366245&partnerID=8YFLogxK
U2 - 10.1016/j.vetimm.2005.01.003
DO - 10.1016/j.vetimm.2005.01.003
M3 - Article
C2 - 15910990
AN - SCOPUS:19344366245
VL - 106
SP - 23
EP - 38
JO - Veterinary Immunology and Immunopathology
JF - Veterinary Immunology and Immunopathology
SN - 0165-2427
IS - 1-2
ER -