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Frontiers in veterinary science2021; 8; 677675; doi: 10.3389/fvets.2021.677675

Effects of Juglone on Neutrophil Degranulation and Myeloperoxidase Activity Related to Equine Laminitis.

Abstract: Experimental laminitis, characterized by a failure of the dermal-epidermal interface of the foot, can be induced in horses by the oral administration of a black walnut extract (BWE). In the early phase of this severe and painful disease, an activation of neutrophil occurs, with the release of myeloperoxidase (MPO), a pro-oxidant enzyme of neutrophils, in plasma, skin, and laminar tissue. Juglone, a naphthoquinone derivative endowed with redox properties, is found in walnuts and has been incriminated in this neutrophil activation. We report for the first time the inhibitory activity of juglone on the degranulation of neutrophils induced by cytochalasin B and formyl-methionyl-leucyl-phenylalanine as monitored by the MPO release (>90% inhibition for 25 and 50 μM). Moreover, it also acts on the peroxidase activity of MPO by interacting with the intermediate "π cation radical," as evidenced by the classical and specific immunological extraction followed by enzymatic detection (SIEFED) assays. These results are confirmed by a docking study showing the perfect positioning of juglone in the MPO enzyme active site and its interaction with one of the amino acids (Arg-239) of MPO apoprotein. By chemiluminescence and electron paramagnetic resonance techniques, we demonstrated that juglone inhibited reactive oxygen species (ROS) and superoxide anion free radical produced from phorbol myristate acetate (PMA)-activated polymorphonuclear neutrophils (PMNs). These results indicate that juglone is not the trigger for equine laminitis, at least if we focus on the modulation of neutrophil activation.
Publication Date: 2021-07-16 PubMed ID: 34336974PubMed Central: PMC8322847DOI: 10.3389/fvets.2021.677675Google Scholar: Lookup
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Summary

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The research study evaluates the effects of a walnut extract compound, Juglone, on the process of neutrophil degranulation and myeloperoxidase activity – elements implicated in the development of a painful horse foot disease, equine laminitis. The study findings suggest that Juglone does not trigger equine laminitis, but rather has inhibitory effects on neutrophil activation.

Research Objective and Background

  • The research aims to explore the role of Juglone, a naphthoquinone derivative and a compound found in walnuts, in neutrophil activation and myeloperoxidase (MPO) activity relating to equine laminitis.
  • Equine laminitis is a severe and painful disease in horses characterised by a failure of the foot’s dermal-epidermal interface. This disease can be induced experimentally in horses by the oral administration of a black walnut extract (BWE).
  • In the early phase of this disease, there is an activation of neutrophil, a type of blood cell involved in the body’s defence against infections, which leads to the release of myeloperoxidase (MPO), a pro-oxidant enzyme of neutrophils, in plasma, skin, and laminar tissue.

Research Findings and Methodology

  • For the first time, the inhibitory activity of juglone on the degranulation of neutrophils was reported in the study. The researchers monitored this by the MPO release and found more than 90% inhibition for 25 and 50 μM.
  • The study further found that juglone also acts on the peroxidase activity of MPO by interacting with the intermediate “π cation radical.” This was validated through classical and specific immunological extraction followed by enzymatic detection (SIEFED) assays.
  • The perfect positioning of juglone in the MPO enzyme active site and its interaction with one of the amino acids (Arg-239) of MPO apoprotein was shown by a docking study.
  • Additionally, through chemiluminescence and electron paramagnetic resonance techniques, it was demonstrated that juglone inhibited reactive oxygen species (ROS) and superoxide anion free radical produced from phorbol myristate acetate (PMA)-activated polymorphonuclear neutrophils (PMNs).

Conclusion and Implication

  • The research concludes that juglone is not the trigger for equine laminitis. This finding challenges the prevailing perception of juglone’s role in the disease, especially considering its presence in the walnut extract that can induce laminitis in horses.
  • The emphasis of juglone’s inhibitory effects on neutrophil activation highlights a potential area of further research: the exploration of juglone or similar compounds as potential therapeutic agents that can modify neutrophil activation and, possibly, deter the development or progression of diseases like equine laminitis.

Cite This Article

APA
Mouithys-Mickalad A, Storms N, Franck T, Ceusters J, de la Rebière de Pouyade G, Deby-Dupont G, Serteyn D. (2021). Effects of Juglone on Neutrophil Degranulation and Myeloperoxidase Activity Related to Equine Laminitis. Front Vet Sci, 8, 677675. https://doi.org/10.3389/fvets.2021.677675

Publication

ISSN: 2297-1769
NlmUniqueID: 101666658
Country: Switzerland
Language: English
Volume: 8
Pages: 677675
PII: 677675

Researcher Affiliations

Mouithys-Mickalad, Ange
  • Centre for Oxygen R&D, Institute of Chemistry, B6a, University of Liège, Liège, Belgium.
Storms, Nazaré
  • Department of Clinical Sciences, Equine Surgery, University of Liège, Liège, Belgium.
Franck, Thierry
  • Centre for Oxygen R&D, Institute of Chemistry, B6a, University of Liège, Liège, Belgium.
Ceusters, Justine
  • Centre for Oxygen R&D, Institute of Chemistry, B6a, University of Liège, Liège, Belgium.
de la Rebière de Pouyade, Geoffroy
  • Department of Clinical Sciences, Equine Surgery, University of Liège, Liège, Belgium.
Deby-Dupont, Ginette
  • Centre for Oxygen R&D, Institute of Chemistry, B6a, University of Liège, Liège, Belgium.
Serteyn, Didier
  • Centre for Oxygen R&D, Institute of Chemistry, B6a, University of Liège, Liège, Belgium.
  • Department of Clinical Sciences, Equine Surgery, University of Liège, Liège, Belgium.

Conflict of Interest Statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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