Biosynthesis of the Novel Endogenous 15-Lipoxygenase Metabolites N-13-Hydroxy-octodecadienoyl-ethanolamine and 13-Hydroxy-octodecadienoyl-glycerol by Human Neutrophils and Eosinophils

  • Cells. 2021 Sep 5;10(9):2322. doi: 10.3390/cells10092322.
Anne-Sophie Archambault  1  2 ,  Francesco Tinto  1  2 ,  Élizabeth Dumais  1  2 ,  Volatiana Rakotoarivelo  1  2 ,  Magdalena Kostrzewa  3 ,  Pier-Luc Plante  4 ,  Cyril Martin  1  2 ,  Mélissa Simard  1  2  5 ,  Cristoforo Silvestri  1  2 ,  Roxane Pouliot  5 ,  Michel Laviolette  1 ,  Louis-Philippe Boulet  1 ,  Rosa Maria Vitale  3 ,  Alessia Ligresti  3 ,  Vincenzo Di Marzo  1  2  3  4  6  7 ,  Nicolas Flamand  1  2
Affiliations
  • 1. Centre de Recherche de l'Institut Universitaire de Cardiologie et de Pneumologie de Québec, Faculté de Médecine, Université Laval, Québec City, QC G1V 4G5, Canada.
  • 2. Canada Excellence Research Chair on the Microbiome-Endocannabinoidome Axis in Metabolic Health (CERC-MEND), Université Laval, Québec City, QC G1V 0A6, Canada.
  • 3. Endocannabinoid Research Group, Institute of Biomolecular Chemistry, Consiglio Nazionale Delle Ricerche (CNR), 80078 Pozzuoli, Italy.
  • 4. Institut sur la Nutrition et les Aliments Fonctionnels, Centre NUTRISS, École de Nutrition, Faculté des Sciences de L'agriculture et de L'alimentation, Université Laval, Québec City, QC G1V 0A6, Canada.
  • 5. Faculté de Pharmacie de l'Université Laval and Centre de Recherche en Organogénèse Expérimentale de l'Université Laval/LOEX, Axe Médecine Régénératrice, Centre de Recherche du CHU de Québec-Université Laval, Québec City, QC G1V 0A6, Canada.
  • 6. Joint International Unit between the Consiglio Nazionale delle Ricerche (CNR), 80078 Pozzuoli, Italy.
  • 7. Canada on Chemical and Biomolecular Research on the Microbiome and Its Impact on Metabolic Health and Nutrition (UMI-MicroMeNu), Université Laval, Québec City, QC G1V 0A6, Canada.
PMID: 34571971 DOI: 10.3390/cells10092322
Abstract

The endocannabinoids 2-arachidonoyl-glycerol and N-arachidonoyl-ethanolamine are lipids regulating many physiological processes, notably inflammation. Endocannabinoid hydrolysis inhibitors are now being investigated as potential anti-inflammatory agents. In addition to 2-arachidonoyl-glycerol and N-arachidonoyl-ethanolamine, the endocannabinoidome also includes Other monoacylglycerols and N-acyl-ethanolamines such as 1-linoleoyl-glycerol (1-LG) and N-linoleoyl-ethanolamine (LEA). By increasing monoacylglycerols and/or N-acyl-ethanolamine levels, endocannabinoid hydrolysis inhibitors will likely increase the levels of their metabolites. Herein, we investigated whether 1-LG and LEA were substrates for the 15-lipoxygenase pathway, given that both possess a 1Z,4Z-pentadiene motif, near their omega end. We thus assessed how human eosinophils and neutrophils biosynthesized the 15-lipoxygenase metabolites of 1-LG and LEA. Linoleic acid (LA), a well-documented substrate of 15-lipoxygenases, was used as positive control. N-13-hydroxy-octodecadienoyl-ethanolamine (13-HODE-EA) and 13-hydroxy-octodecadienoyl-glycerol (13-HODE-G), the 15-lipoxygenase metabolites of LEA and 1-LG, were synthesized using Novozym 435 and soybean Lipoxygenase. Eosinophils, which express the 15-lipoxygenase-1, metabolized LA, 1-LG, and LEA into their 13-hydroxy derivatives. This was almost complete after five minutes. Substrate preference of eosinophils was LA > LEA > 1-LG in presence of 13-HODE-G hydrolysis inhibition with methyl-arachidonoyl-fluorophosphonate. Human neutrophils also metabolized LA, 1-LG, and LEA into their 13-hydroxy derivatives. This was maximal after 15-30 s. Substrate preference was LA ≫ 1-LG > LEA. Importantly, 13-HODE-G was found in humans and mouse tissue samples. In conclusion, our data show that human eosinophils and neutrophils metabolize 1-LG and LEA into the novel endogenous 15-lipoxygenase metabolites 13-HODE-G and 13-HODE-EA. The full biological importance of 13-HODE-G and 13-HODE-EA remains to be explored.

Keywords
13-HODE; 2-arachidonoyl-glycerol; N-linoleoyl-ethanolamine; anandamide; eicosanoid; endocannabinoid; eosinophils; linoleic acid; linoleoyl-glycerol; neutrophils.
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