Modulation of equine articular chondrocyte messenger RNA levels following brief exposures to recombinant equine interleukin-1β

V. A. Takafuji, R. D. Howard, D. L. Ward, L. V. Sharova, M. V. Crisman

Research output: Contribution to journalArticlepeer-review

6 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)23-38
Number of pages16
JournalVeterinary Immunology and Immunopathology
Volume106
Issue number1-2
DOIs
StatePublished - Jun 15 2005
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Immunology
  • veterinary(all)

Keywords

  • Chondrocytes
  • Gene expression
  • Interleukin-1β
  • Osteoarthritis

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