Modulation of equine articular chondrocyte messenger RNA levels following brief exposures to recombinant equine interleukin-1beta.
Abstract: The effect of recombinant equine IL-1beta (EqIL-1beta) 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-1beta 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 > or =two-fold in response to EqIL-1beta after 1, 3 and 6h (1, 2 and 109 transcripts, respectively). At 6 h of EqIL-1beta 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 ( approximately 10% of the total variation) corresponding to a potential EqIL-1beta 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.
Publication Date: 2005-05-25 PubMed ID: 15910990DOI: 10.1016/j.vetimm.2005.01.003Google Scholar: Lookup
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- Journal Article
Summary
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This research examined how the protein equine interleukin-1beta (EqIL-1beta) impacts the gene expression levels of horse joint cells (chondrocytes), which might be related to the development of horse osteoarthritis. The study found that exposure to EqIL-1beta resulted in a time-dependent increase in expressed transcripts (gene products), especially associated with cell function and inflammatory processes.
Objectives and Methodology
- This study was conducted to understand the impact of a recombinant protein, EqIL-1beta, on chondrocytes – the cells found in cartilage tissue in horse joints.
- Cells were cultured in a media with and without EqIL-1beta for various periods. The researchers aimed to understand how this protein changes the cells’ genetic activity over time.
- The scientists used a specialized cDNA array to analyze the cells’ activity. This means they were measuring the amount of messenger RNA (mRNA) – the intermediate step between genes and proteins they encode.
- The fluctuating mRNA levels indirectly indicate changes in gene activity, allowing the researchers to observe any variations in behavior caused by the presence of EqIL-1beta.
Main Findings
- The researchers detected a trend – over time, the number of transcripts that were upregulated (or produced in larger quantities) in response to EqIL-1beta increased.
- After 6 hours, 88 particular gene sequences had increased expression levels by more than two-fold. These genes are associated with various cell functions, such as cell cycle, intracellular signaling, transcription, translation, extracellular matrix turnover, and inflammation.
- Through principal component analysis, the researchers identified a potential co-regulation of EqIL-1beta and cell cycle related gene transcription.
Conclusions
- The study broadens our understanding of how EqIL-1beta modulates gene expression in chondrocytes, especially with regards to inflammatory processes.
- These insights suggest that IL-1 is involved, either directly or indirectly, in the development of inflammation that drives diseases like osteoarthritis in horse joints.
- The study encourages further research on the molecular events underlying inflammation and joint disease development due to IL-1.
Cite This Article
APA
Takafuji VA, Howard RD, Ward DL, Sharova LV, Crisman MV.
(2005).
Modulation of equine articular chondrocyte messenger RNA levels following brief exposures to recombinant equine interleukin-1beta.
Vet Immunol Immunopathol, 106(1-2), 23-38.
https://doi.org/10.1016/j.vetimm.2005.01.003 Publication
Researcher Affiliations
- Orthopedic Research Laboratory, VA-MD Regional College of Veterinary Medicine, Blacksburg, VA 24061-0442, USA.
MeSH Terms
- Animals
- Cartilage, Articular / drug effects
- Cartilage, Articular / metabolism
- Cells, Cultured
- Chondrocytes / drug effects
- Chondrocytes / metabolism
- Gene Expression Regulation / drug effects
- Horses / metabolism
- Interleukin-1 / pharmacology
- Interleukin-1 / physiology
- Oligonucleotide Array Sequence Analysis / veterinary
- RNA, Messenger / metabolism
- Recombinant Proteins / pharmacology
- Time Factors
Citations
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