Inducible nitric oxide expression in equine articular chondrocytes: effects of antiinflammatory compounds.
Abstract: To determine the effects of recombinant equine IL-1beta and a number of antiinflammatory compounds on the expression and activity of inducible nitric oxide synthase (iNOS) in cultured equine chondrocytes. Methods: RT-PCR methods were used to amplify a portion of the equine iNOS message to prepare an RNA probe. Northern blot analysis was used to quantify the expression of iNOS in first passage cultures of equine articular chondrocytes propagated in the presence or absence of recombinant equine interleukin-1beta (reIL-1beta), dexamethasone (DEX), polysulfated glycosaminoglycan (PSGAG), hyaluronan (HA), and phenylbutazone (PBZ), each at concentrations of 10 and 100 microg/ml. Nitrite concentrations in conditioned media of similarly treated cells were used to quantify iNOS activity. Results: Recombinant equine IL-1beta increased the expression of iNOS in a dose-dependent manner. This result was paralleled by an increased concentration of nitrite in the culture media of reIL-1beta-treated cells. DEX and PSGAG significantly reduced iNOS gene expression and media supernatant nitrite concentrations in cytokine-stimulated cultures. HA and PBZ had no consistent effect on the expression of iNOS and did not significantly influence nitrite content of conditioned media. Conclusions: NO is considered an important mediator in the pathophysiologic processes of arthritis and an inducible NOS is expressed by equine chondrocytes. Pre-translational regulation of the iNOS gene by DEX and PSGAG appears to contribute to the cartilage-sparing properties of these compounds.
Copyright 2002 OsteoArthritis Research Society International.
Publication Date: 2002-02-09 PubMed ID: 11795978DOI: 10.1053/joca.2001.0476Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research investigated how certain anti-inflammatory compounds affect inducible nitric oxide synthase (iNOS) expression and activity in horse joint cells. It found that recombinant equine IL-1beta boosts iNOS expression, and that dexamethasone and polysulfated glycosaminoglycan can reduce it.
Methods
- The scientists used RT-PCR methods to amplify a part of the equine iNOS message to prepare an RNA probe. This technology is a common way to amplify specific parts of genes to study their behaviors in cells.
- They used Northern blot analysis to measure the expression of iNOS in first passage culture of horse joint cells propagated in the presence or absence of several compounds. Northern blot is a technique used to detect specific RNA molecules among a mixture of RNA.
- The compounds they investigated were recombinant equine interleukin-1beta (reIL-1beta), dexamethasone (DEX), polysulfated glycosaminoglycan (PSGAG), hyaluronan (HA), and phenylbutazone (PBZ), each tested at concentrations of 10 and 100 microg/ml.
- The research team used the nitrite concentrations in conditioned media of similarly treated cells to quantify iNOS activity. Nitrite concentrations are often used as an indicator of iNOS enzyme activity as iNOS catalyzes the conversion of one form of nitrate to nitrite.
Results
- Recombinant equine IL-1beta led to an increase in the expression of iNOS in a dose-dependent manner. They also found higher nitrite levels in the culture media of cells treated with this compound.
- DEX and PSGAG significantly reduced iNOS gene expression and media supernatant nitrite concentrations in cytokine-stimulated cultures. This indicates that these anti-inflammatory compounds have a positive effect in reducing inflammation by reducing the activity of iNOS.
- HA and PBZ did not have a consistent effect on the expression of iNOS and did not significantly affect nitrite content of conditioned media. This suggests that these compounds are less effective in controlling the inflammation response.
Conclusions
- Nitric oxide is considered an important player in the development of arthritis, and the fact that an inducible NOS is expressed by horse joint cells supports its role in this disease.
- The control of the iNOS gene by DEX and PSGAG before translation seems to contribute to the protective effects these compounds have on cartilage, making them potentially effective treatments for conditions like arthritis.
Cite This Article
APA
Tung JT, Venta PJ, Caron JP.
(2002).
Inducible nitric oxide expression in equine articular chondrocytes: effects of antiinflammatory compounds.
Osteoarthritis Cartilage, 10(1), 5-12.
https://doi.org/10.1053/joca.2001.0476 Publication
Researcher Affiliations
- Department of Large Animal Clinical Sciences, Michigan State University, East Lansing, Michigan 48824-1314, USA.
MeSH Terms
- Adjuvants, Immunologic / pharmacology
- Analysis of Variance
- Animals
- Anti-Inflammatory Agents / pharmacology
- Anti-Inflammatory Agents, Non-Steroidal / pharmacology
- Blotting, Northern
- Cartilage, Articular / cytology
- Cartilage, Articular / drug effects
- Cartilage, Articular / metabolism
- Cells, Cultured
- Chondrocytes / drug effects
- Chondrocytes / metabolism
- Dexamethasone / pharmacology
- Dose-Response Relationship, Drug
- Glycosaminoglycans / pharmacology
- Horses / physiology
- Hyaluronic Acid / pharmacology
- Interleukin-1 / physiology
- Mass Spectrometry
- Nitric Oxide Synthase / drug effects
- Nitric Oxide Synthase / metabolism
- Nitric Oxide Synthase Type II
- Phenylbutazone / pharmacology
- RNA Probes
- Reverse Transcriptase Polymerase Chain Reaction
Citations
This article has been cited 11 times.- Taguchi T, Morales Yniguez FJ, Takawira C, Andrews FM, Lopez MJ. Agmatine Administration Effects on Equine Gastric Ulceration and Lameness. J Clin Med 2022 Dec 8;11(24).
- Kim YS, Suh DS, Tak DH, Chung PK, Kwon YB, Kim TY, Koh YG. Comparative matched-pair cohort analysis of the short-term clinical outcomes of mesenchymal stem cells versus hyaluronic acid treatments through intra-articular injections for knee osteoarthritis. J Exp Orthop 2020 Nov 13;7(1):90.
- Bwalya EC, Kim S, Fang J, Wijekoon HMS, Hosoya K, Okumura M. Pentosan polysulfate inhibits IL-1β-induced iNOS, c-Jun and HIF-1α upregulation in canine articular chondrocytes. PLoS One 2017;12(5):e0177144.
- Wernecke C, Braun HJ, Dragoo JL. The Effect of Intra-articular Corticosteroids on Articular Cartilage: A Systematic Review. Orthop J Sports Med 2015 May;3(5):2325967115581163.
- Parkinson L, Keast R. Oleocanthal, a phenolic derived from virgin olive oil: a review of the beneficial effects on inflammatory disease. Int J Mol Sci 2014 Jul 11;15(7):12323-34.
- Carrade DD, Lame MW, Kent MS, Clark KC, Walker NJ, Borjesson DL. Comparative Analysis of the Immunomodulatory Properties of Equine Adult-Derived Mesenchymal Stem Cells(). Cell Med 2012;4(1):1-11.
- Chou LW, Wang J, Chang PL, Hsieh YL. Hyaluronan modulates accumulation of hypoxia-inducible factor-1 alpha, inducible nitric oxide synthase, and matrix metalloproteinase-3 in the synovium of rat adjuvant-induced arthritis model. Arthritis Res Ther 2011 Jun 16;13(3):R90.
- Kalaci A, Yilmaz HR, Aslan B, Söğüt S, Yanat AN, Uz E. Effects of hyaluronan on nitric oxide levels and superoxide dismutase activities in synovial fluid in knee osteoarthritis. Clin Rheumatol 2007 Aug;26(8):1306-11.
- Moreland LW. Intra-articular hyaluronan (hyaluronic acid) and hylans for the treatment of osteoarthritis: mechanisms of action. Arthritis Res Ther 2003;5(2):54-67.
- Pirri C, Sorbino A, Manocchio N, Pirri N, Devito A, Foti C, Migliore A. Chondrotoxicity of Intra-Articular Injection Treatment: A Scoping Review. Int J Mol Sci 2024 Jun 26;25(13).
- Ibad HA, Kasaeian A, Ghotbi E, Roemer F, Jarraya M, Ghazi-Sherbaf F, Dolatshahi M, Demehri S, Guermazi A. Longitudinal MRI-defined Cartilage Loss and Radiographic Joint Space Narrowing Following Intra-Articular Corticosteroid Injection for Knee Osteoarthritis: A Systematic Review and Meta-analysis. Osteoarthr Imaging 2023 Sep;3(3).
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