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Dose-dependent effects of corticosteroids on the expression of matrix-related genes in normal and cytokine-treated articular chondrocytes.
Abstract: To assess the effects of glucocorticoids on the expression of multiple matrix-related genes in normal and cytokine-treated cultured equine articular chondrocytes in a phenotypically correct suspension culture. Methods: Articular cartilage harvested from the joints of 15 foals, 7 yearling horses, and 16 adult horses. Methods: Glucocorticoids (dexamethasone, prednisolone, triamcinolone) at 10(-10) to 10(-4) M. Methods: Equine articular chondrocytes maintained in suspension cultures were treated with glucocorticoids with and without human recombinant interleukin 1-beta (IL1-beta) and tumor necrosis factor-alpha (TNF-alpha). Northern blots of total RNA from the treated cells were probed with equine specific cDNA probes for a number of cartilage matrix-related genes. Zymography, Western blotting, and fluorography were also performed to study the effects on protein synthesis. Results: The glucocorticoids, dexamethasone, triamcinolone, and prednisolone, markedly decreased MMP1, MMP3, MMP13, TIMPI, and ferritin steady-state mRNA levels. There were no qualitative differences seen among the tested corticosteroids although dexamethasone and triamcinolone appeared to be slightly more potent than prednisolone. The effects of the glucocorticoids on MMP transcription occurred consistently at lower doses than those required to similarly downregulate type II collagen and aggrecan. Link protein and fibronectin mRNA were increased by the glucocorticoids, and biglycan and decorin were minimally affected. Fluorography of [14-C] proline-labeled media demonstrated that the decrease in type II collagen transcription (mRNA levels) was paralleled at the protein level. Zymography and Western blotting confirmed the decrease in functional metalloproteinases found in chondrocyte cultures following glucocorticoid treatment. Conclusions: The effects of glucocorticoids are complex inasmuch as they differentially affect numerous genes involved in the composition of cartilage matrix and the degradation of that matrix. This study provides new insight into the effects of glucocorticoids on the regulation of extra-cellular matrix and matrix-related genes by demonstrating that low doses of glucocorticoids can inhibit the degradative metalloproteinases with minimal negative effects on the transcription of extracellular matrix genes.
Publication Date: 2003-03-01 PubMed ID: 12608648DOI: 10.1007/s000110300012Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
- Research Support
- Non-U.S. Gov't
- Research Support
- U.S. Gov't
- P.H.S.
Summary
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The study investigates the dose-dependent effects of glucocorticoids on the gene expression of several matrix-associated components in cultured horse joint chondrocytes, with and without cytokine treatments. It found that glucocorticoids affect the expression of these genes differently, and lower doses can hamper the destructive metalloproteinases with limited negative impact on the transcription of genes involved in producing the extracellular matrix.
Research Objectives and Methodology
- The primary goal of this study was to understand how glucocorticoids—an anti-inflammatory medication—affect the expression of various genes related to the extracellular matrix in horse chondrocytes, which are cells within the cartilage tissue. The researchers also aimed to investigate these effects under normal conditions and when these cells are exposed to cytokines, a type of immune signaling molecule.
- The chondrocyte cells for the research were sourced from the joints of foals, yearlings, and adult horses, and maintained in a suspension culture, meaning they were grown in a liquid medium.
- Three different glucocorticoids were administered at varying concentrations to these chondrocytes, namely, dexamethasone, triamcinolone, and prednisolone. For the cytokine-treated group, these glucocorticoids were co-treated with the cytokines interleukin 1-beta (IL1-beta) and tumor necrosis factor-alpha (TNF-alpha).
- The team used several techniques, including Northern blotting, zymography, fluorography and Western blotting, to examine the effects of these treatments on the expression of many matrix-related genes and their protein synthesis.
Research Findings
- The study revealed that all three glucocorticoids reduced the expression of the MMP1, MMP3, MMP13, TIMPI, and ferritin genes. Though no qualitative differences were observed among the glucocorticoids, dexamethasone and triamcinolone were slightly more potent than prednisolone.
- Interestingly, lower glucocorticoid doses were needed to consistently impact the transcription of MMPs than to decrease type II collagen and aggrecan, two major components of cartilage tissue.
- The mRNA levels of link protein and fibronectin, essential players in maintaining the structural integrity of the cartilage, were increased by glucocorticoids, and two other constituents, biglycan and decorin, were marginally affected.
- Decrease in type II collagen transcription (mRNA levels) paralleled the protein level as shown by the fluorography study.
- Both zymography and Western blot tests confirmed a decrease in functional metalloproteinases, which are enzymes that degrade the matrix, post glucocorticoid treatment.
Conclusions
- The study concludes that glucocorticoids have multifaceted effects on gene expressions that determine the composition and degradation of cartilage tissue. They selectively impact various genes involved in the process.
- The findings further highlight that lower doses of glucocorticoids can deter the expression of cartilage-destructive metalloproteinases with minimal negative impact on the transcription of genes contributing to the production of extracellular matrix.
- This study thus provides crucial insights into the potential benefits and limitations of glucocorticoids in treating osteoarthritis and similar conditions characterized by degradation of cartilage.
Cite This Article
APA
Richardson DW, Dodge GR.
(2003).
Dose-dependent effects of corticosteroids on the expression of matrix-related genes in normal and cytokine-treated articular chondrocytes.
Inflamm Res, 52(1), 39-49.
https://doi.org/10.1007/s000110300012 Publication
Researcher Affiliations
- Department of Clinical Studies, New Bolton Center, School of Veterinary Medicine, University of Pennsylvania, Kennett Square, PA 19348-1692, USA. dwr@vet.upenn.edu
MeSH Terms
- Adrenal Cortex Hormones / pharmacology
- Animals
- Blotting, Northern
- Blotting, Western
- Bone Matrix / physiology
- Cartilage, Articular
- Cells, Cultured
- Chondrocytes / drug effects
- Chondrocytes / metabolism
- Cytokines / pharmacology
- Dexamethasone / pharmacology
- Dose-Response Relationship, Drug
- Ferritins / biosynthesis
- Gene Expression Regulation / drug effects
- Glucocorticoids / pharmacology
- Horses
- Humans
- Interleukin-1 / pharmacology
- Matrix Metalloproteinases / biosynthesis
- Matrix Metalloproteinases / genetics
- Phenotype
- Prednisolone / pharmacology
- Protein Biosynthesis
- Proteins / genetics
- RNA, Messenger / biosynthesis
- Recombinant Proteins / pharmacology
- Triamcinolone / pharmacology
- Tumor Necrosis Factor-alpha / pharmacology
Grant Funding
- AR42417 / NIAMS NIH HHS
Citations
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