Effects of adenosine on bacterial lipopolysaccharide- and interleukin 1-induced nitric oxide release from equine articular chondrocytes.
Abstract: To determine whether adenosine influences the in vitro release of nitric oxide (NO) from differentiated primary equine articular chondrocytes. Methods: Articular cartilage harvested from the metacarpophalangeal and metatarsophalangeal joints of 11 horses (3 to 11 years old) without history or clinical signs of joint disease. Methods: Chondrocytes were isolated, plated at a high density (10(5) cells/well), and treated with adenosine, the adenosine receptor agonist 5'-N-ethylcarboxamidoadenosine (NECA), bradykinin, or other agents that modify secondary messenger pathways alone or in combination with bacterial lipopolysaccharide (LPS) or recombinant human interleukin-1alpha (rhIL-1alpha). Nitric oxide release was measured indirectly by use of the Griess reaction and was expressed as micromol of nitrite in the supernatant/microg of protein in the cell layer. Inducible nitric oxide synthase (iNOS) activity was determined by measuring the conversion of radiolabeled arginine to radiolabeled citrulline. Results: Treatment of chondrocytes with adenosine alone had no significant effect on NO release. However, adenosine and NECA inhibited LPS- and rhIL-1alpha-induced NO release. This response was mimicked by forskolin, which acts to increase adenylate cyclase activity, but not by the calcium ionophore A23187 Treatment of chondrocytes with phorbol myristate acetate, which acts to increase protein kinase C activity, potentiated LPS-induced NO release. Adenosine treatment also significantly inhibited the LPS-induced increase in iNOS activity. Conclusions: Adenosine and the nonspecific adenosine receptor agonist NECA inhibited inflammatory mediator-induced release of NO from equine articular chondrocytes. Modulation of adenosine receptor-mediated pathways may offer novel methods for treatment of inflammation in horses with joint disease.
Publication Date: 2002-02-15 PubMed ID: 11843119DOI: 10.2460/ajvr.2002.63.204Google 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
- U.S. Gov't
- P.H.S.
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research investigated the role adenosine plays in the release of nitric oxide (NO) from equine joint cells. The study found that adenosine and an adenosine receptor agonist can inhibit the release of NO triggered by inflammatory mediators, potentially suggesting new approaches for treating inflammation in horse joint disorders.
Design and Methods of the Study
- The research involved articular cartilage obtained from the metacarpophalangeal and metatarsophalangeal joints of 11 horses aged between 3 and 11 years old, with no prior history or clinical indications of joint diseases.
- Chondrocytes, the primary cells in the cartilage, were separated, plated in high density, and treated with different agents that include adenosine, the adenosine receptor agonist 5′-N-ethylcarboxamidoadenosine (NECA), bradykinin, among other agents influencing secondary messenger pathways.
- The treatments were done separately or in conjunction with bacterial lipopolysaccharide (LPS) or recombinant human interleukin-1alpha (rhIL-1alpha), which are known to cause inflammation.
Procedure for Measuring Nitric Oxide (NO) Release, and iNOS Activity
- Nitric oxide release was indirectly measured using the Griess reaction, indicating the amount of nitrite present in the cell supernatant to the protein content in the cell layer.
- Inducible nitric oxide synthase (iNOS) activity, which is associated with the production of NO, was determined by observing the conversion of radioactively labelled arginine to citrulline.
Findings and Conclusions
- No significant effect on nitric oxide release was noticed when chondrocytes were treated solely with adenosine. In contrast, both adenosine and NECA were able to inhibit the release of NO induced by LPS and rhIL-1alpha.
- Adenosine treatment considerably inhibited the LPS-induced increase in iNOS activity.
- These conclusions suggest that adenosine and NECA could act as inhibitors for the release of NO, induced by activating inflammatory pathways from equine articular chondrocytes.
- Research points to a potential opportunity where adenosine receptor-mediated pathways may be used as novel methods for the treatment of inflammation in horses with joint disease.
Cite This Article
APA
Benton HP, MacDonald MH, Tesch AM.
(2002).
Effects of adenosine on bacterial lipopolysaccharide- and interleukin 1-induced nitric oxide release from equine articular chondrocytes.
Am J Vet Res, 63(2), 204-210.
https://doi.org/10.2460/ajvr.2002.63.204 Publication
Researcher Affiliations
- Department of Anatomy, Physiology and Cell Biology, School of Veterinary Medicine, University of California, Davis 95616, USA.
MeSH Terms
- Adenosine / pharmacology
- Adenosine-5'-(N-ethylcarboxamide) / pharmacology
- Animals
- Bradykinin / pharmacology
- Calcimycin / pharmacology
- Cartilage, Articular / drug effects
- Cartilage, Articular / enzymology
- Cells, Cultured
- Chondrocytes / drug effects
- Chondrocytes / enzymology
- Drug Interactions
- Horses / metabolism
- Interleukin-1 / pharmacology
- Ionophores / pharmacology
- Lipopolysaccharides / pharmacology
- Nitric Oxide Synthase / metabolism
- Nitric Oxide Synthase Type II
- Protein Kinase C / metabolism
- Purinergic P1 Receptor Agonists
Grant Funding
- AR41475 / NIAMS NIH HHS
Citations
This article has been cited 7 times.- Ghonimy A, Chen Z, Li J. The effect of C/N ratio and its frequent addition on commensal and pathogenic bacterial abundances in shrimp Litopeaneus vanname gut in a biofloc system: Ratio and frequent addition interaction matters. PLoS One 2023;18(4):e0283841.
- Corciulo C, Castro CM, Coughlin T, Jacob S, Li Z, Fenyö D, Rifkin DB, Kennedy OD, Cronstein BN. Intraarticular injection of liposomal adenosine reduces cartilage damage in established murine and rat models of osteoarthritis. Sci Rep 2020 Aug 10;10(1):13477.
- Corciulo C, Cronstein BN. Signaling of the Purinergic System in the Joint. Front Pharmacol 2019;10:1591.
- Bekisz JM, Lopez CD, Corciulo C, Mediero A, Coelho PG, Witek L, Flores RL, Cronstein BN. The Role of Adenosine Receptor Activation in Attenuating Cartilaginous Inflammation. Inflammation 2018 Aug;41(4):1135-1141.
- Corciulo C, Lendhey M, Wilder T, Schoen H, Cornelissen AS, Chang G, Kennedy OD, Cronstein BN. Endogenous adenosine maintains cartilage homeostasis and exogenous adenosine inhibits osteoarthritis progression. Nat Commun 2017 May 11;8:15019.
- Chua NH, Halim W, Evers AW, Vissers KC. Whiplash patients with cervicogenic headache after lateral atlanto-axial joint pulsed radiofrequency treatment. Anesth Pain Med 2012 Winter;1(3):162-7.
- Chua NH, Vissers KC, Sluijter ME. Pulsed radiofrequency treatment in interventional pain management: mechanisms and potential indications-a review. Acta Neurochir (Wien) 2011 Apr;153(4):763-71.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
