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Home / Equine Research Bank / Arthritis Res Ther / A targeted lipidomics approach to the study of eicosanoid re...
Arthritis research & therapy2011; 13(4); R123; doi: 10.1186/ar3427

A targeted lipidomics approach to the study of eicosanoid release in synovial joints.

Abstract: Articular tissues are capable of producing a range of eicosanoid mediators, each of which has individual biological effects and may be affected by anti-inflammatory treatment. We set out to develop and evaluate a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) approach for the simultaneous analysis of multiple eicosanoid lipid mediators in equine synovial fluid (SF), and to illustrate its use for investigation of the in vivo effects of non-steroidal anti-inflammatory drug (NSAID) treatment. Methods: Synovial fluid samples were obtained from normal joints of 6 adult horses at baseline (0 hr) and at 8, 24 and 168 hours after experimental induction of transient acute synovitis, with horses treated once daily with oral NSAID (meloxicam, 0.6 mg/kg) or placebo. Following solid-phase extraction, SF lipid mediator quantitation was based on liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) analysis, and results were compared between disease states using linear discriminant analysis (LDA) and analysis of variance (ANOVA) with multiple comparisons corrections. Results: Of a total of 23 mediators targeted, 14 could be reliably identified and quantified in SF samples based on detection of characteristic fragment ions at retention times similar to those of commercial standards. LDA analysis of baseline, 8, 24 and 168 hour synovial fluid samples revealed a separation of these groups into discrete clusters, reflecting dynamic changes in eicosanoid release over the course of synovitis. Prostaglandin (PG) E(2) was significantly lower in NSAID vs. placebo treated samples at all time points; PGE(1), 11-hydroxyeicosatetraenoic acid (11-HETE) and 13,14-dihydro-15keto PGF(2)α were reduced at 8 and 24 hours by NSAID treatment; while 15-HETE, 6-keto PGF(1)α, PGF(2)α, 13,14-dihydro-15keto PGE(2) and thromboxane B(2) (TXB(2)) were reduced at the 8 hour time point only. An interesting pattern was seen for Leukotriene B(4) (LTB(4)), NSAID treatment causing an initial increase at 8 hours, but a significant reduction by 168 hours. Conclusions: The described method allows a comprehensive analysis of synovial fluid eicosanoid profiles. Eicosanoid release in inflamed joints as well as differences between NSAID treated and placebo treated individuals are not limited to PGE(2) or to the early inflammatory phase.
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Publication Date: 2011-07-27 PubMed ID: 21794148PubMed Central: PMC3239362DOI: 10.1186/ar3427Google Scholar: Lookup
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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.

This research investigates the effect of non-steroidal anti-inflammatory drugs (NSAIDs) on the production of eicosanoids, a group of compounds involved in inflammation, in the joints of horses. A new method using high-performance liquid chromatography combined with mass spectrometry (HPLC-MS/MS) was applied to measure multiple eicosanoids at once in synovial fluid, showing how eicosanoid release changes over time and in response to NSAID treatment.

Development of HPLC-MS/MS Method for Eicosanoid Analysis

  • The researchers designed a high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) approach to analyse multiple eicosanoid lipid mediators at a time in equine synovial fluid.
  • This method required the collection of synovial fluid from healthy horses at different time points following the experimental induction of synovitis, allowing researchers to track changes in the production of eicosanoids over time.
  • The procedure involved a solid-phase extraction to purify the fluid before identifying and quantifying the eicosanoids based on their distinctive fragment ions and retention times compared to known standards.

Evaluation of Eicosanoid Release

  • Analysis showed that of the 23 mediators targeted for detection, 14 could be consistently identified and quantified in the samples.
  • These results were able to separate different disease states into distinct groups, indicating significant changes in eicosanoid release at various stages of synovitis.
  • Use of NSAID treatment also affected these levels at certain time points, with a range of different eicosanoids showing reductions compared to placebo-treated samples.

Effects of NSAID Treatment

  • The researchers found that certain eicosanoids, such as Prostaglandin (PG) E(2), were continually lower in NSAID-treated samples at all time points.
  • Other molecules like PGE(1), 11-hydroxyeicosatetraenoic acid (11-HETE), and 13,14-dihydro-15keto PGF(2)α were reduced at 8 and 24 hours following NSAID treatment.
  • An interesting effect was observed with Leukotriene B(4) (LTB(4)), which saw an initial increase at 8 hours, but was significantly reduced by 168 hours after NSAID treatment.

Conclusion

  • The proposed HPLC-MS/MS method provides a comprehensive analysis of eicosanoid profiles in synovial fluid, demonstrating how these compounds change dynamically over the course of inflammation and in response to NSAID treatment.
  • The findings indicate that the effects of NSAID treatment on eicosanoid release isn’t limited to the early stages of inflammation or to only certain types of eicosanoids.

Cite This Article

APA
de Grauw JC, van de Lest CH, van Weeren PR. (2011). A targeted lipidomics approach to the study of eicosanoid release in synovial joints. Arthritis Res Ther, 13(4), R123. https://doi.org/10.1186/ar3427

Publication

Arthritis research & therapy
ISSN: 1478-6362
NlmUniqueID: 101154438
Country: England
Language: English
Volume: 13
Issue: 4
Pages: R123

Researcher Affiliations

de Grauw, Janny C
  • Department of Equine Sciences, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 114, 3584 CM, Utrecht, The Netherlands. j.c.degrauw@uu.nl
van de Lest, Chris H A
    van Weeren, Paul René

      MeSH Terms

      • Animals
      • Anti-Inflammatory Agents, Non-Steroidal / therapeutic use
      • Chromatography, High Pressure Liquid / methods
      • Eicosanoids / analysis
      • Eicosanoids / metabolism
      • Horses
      • Synovial Fluid / chemistry
      • Synovitis / drug therapy
      • Synovitis / metabolism
      • Tandem Mass Spectrometry / methods

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