Influence of N-acyl-ethanolamine of arachidonic acid on the synthesis of cytokines and oxylipins by the blood leukocytes of patients with asthma under in vitro conditions

   Introduction. The search for hihly effective methods for the treatment and control of asthma remains an urgent task of modern pathophysiology and pharmacology. N-acyl-ethanolamines of arachidonic acid (NAE 20: 4) are promising biomolecules for the regulation of the immune processes.

   Aim. To investigate the in vitro influence of NAE 20: 4 on the immune cells' ability from patients with asthma to synthesize pro-inflammatory cytokines and oxylipins.

   Materials and methods. The objective of the study was the blood of 11 patients with mild controlled asthma and 10 healthy volunteers. The in vitro experiment was carried out in stimulated and not stimulated by lipopolysaccharide (LPS) blood for 6 hours. NAE 20: 4 was added at 1.0 µM, 3.0 µM or 10.0 µM doses. The levels of cytokines (TNFα, IL-8), thromboxane
B2 (TXB2) and leukotriene B4 (LTB4) were analyzed by the ELISA method.

   Results. We found an increase in TNFα, IL-8, TXB2, LTB4 in the blood of persons with asthma before in vitro NAE 20: 4 exposure, which indicates systemic chronic inflammation. Incubation of blood cells without LPS did not affect the change in the spectrum of mediators studied. Whereas the treatment of NAE 20: 4 at a 10 µM reduced the synthesis of TNFα, IL-8, LTB4, TXB2. In vitro blood cells stimulation by LPS led to hyperproduction of cytokines and oxylipins. NAE 20: 4 at a dose of 1 µM in LPS-stimulated blood had no effect on the production of signaling molecules, while NAE 20:4 at a concentration of 3 µM reduced the level of TNFα, IL-8. NAE 20:4 at a dose of 10.0 µM inhibited the cytokines and LTB4 synthesis. The level of TXB2 under
the influence of NAE 20: 4 at 10.0 µM did not change.

   Conclusion. The results of the study revealed a dose-dependent anti-inflammatory effect of NAE 20:4, characterized by its ability to influence the synthesis of pro-inflammatory cytokines and oxylipins by the immune cells of individuals with asthma in vitro. Further study of NAE 20: 4 opens up new prospects for the development of targeted methods for regulating immune processes in asthma.

1. Global Initiative for Asthma (GINA). Global Strategy for Asthma Management and Prevention, 2018. URL: https://ginasthma.org

2. Papi A., Brightling C., Pedersen S. E., Reddel H. K. Asthma // Lancet. 2018. Vol. 391, Iss.10122. P. 783−800. https://doi.org/10.1016/S0140-6736(17)33311-1

3. Tsabouri S., Mavroudi A., Feketea G., Guibas G. V. Subcutaneous and sublingual immunotherapy in allergic asthma in children // Front. Pediatr. 2017. Vol.5. Article number: 8. https://doi.org/10.3389/fped.2017.00187

4. Berdine G., Alexander R., Nugent K. Clinical entities, phenotypes, causation, and endotypes based on selected asthma publications // Proc. (Bayl. Univ. Med. Cent.). 2020. Vol. 33, Iss.4. P.580−585. https://doi.org/10.1080/08998280.2020.1793444.

5. Patel D., Witt S. N. Ethanolamine and Phosphatidylethanolamine: Partners in Health and Disease // Oxid. Med. Cell. Longev. 2017. Vol.2017. Article number: 4829180. https://doi.org/10.1155/2017/4829180

6. Paton K. F., Shirazi R., Vyssotski M., Kivell B. M. N-docosahexaenoyl ethanolamine (synaptamide) has antinociceptive effects in male mice // Eur. J. Pain. 2020. Vol. 24, Iss.10. P.1990−1998. https://doi.org/10.1002/ejp.1641 2020

7. Meijerink J., Plastin P., Vincken J.P., Poland M., Attya M., Balvers M., Gruppen H., Gabriele B., Witkamp R. F. The ethanolamide metabolite of DHA, docosahexaenoylethanolamine, shows immunomodulating effects in mouse peritoneal and RAW264.7 macrophages: evidence for a new link between fish oil and inflammation // Br. J. Nutr. 2011. Vol.105, Iss.12. Р.1798–1807. https://doi.org/10.1017/S0007114510005635

8. Shirazi R. S., Vyssotski M., Lagutin K., Thompson D., MacDonald Ch., Luscombe V., Glass M., Parker K., Gowing E. K., Williams D., Clarkson A. N. Neuroprotective activity of new Δ3-N-acylethanolamines in a focal ischemia stroke model // Lipids. 2021. Vol. 57, Iss.1. P. 1–15. https://doi.org/10.1002/lipd.12326

9. Ghanbari M. M., Loron A. G., Sayyah M. The ω-3 endocannabinoid docosahexaenoyl ethanolamide reduces seizure susceptibility in mice by activating cannabinoid type 1 receptors // Brain Res. Bull. Vol. 2021. Vol.170. P. 74−80. https://doi.org/10.1016/j.brainresbull.2021.02.011

10. Meijerink J., Poland M., Balvers M. G., Plastina P., Lute C., Dwarkasing J., van Norren K., Witkamp R. F. Inhibition of COX-2-mediated eicosanoid production plays a major role in the anti-inflammatory effects of the endocannabinoid N-docosahexaenoylethanolamine (DHEA) in macrophages // Br. J. Pharmacol. 2014. Vol. 172 (1): 24−37. https://doi.org/10.1111/bph.12747

11. Kytikova O., Novgorodtseva T., Antonyuk M., Denisenko Y., Gvozdenko T. Molecular Targets of Fatty Acid Ethanolamides in Asthma // Medicina. 2019. Vol.55, Iss. 4. Article number: 87. https://doi.org/10.3390/medicina55040087

12. Karaman Yu. K., Lobanova E. G. [Endocannabinoids and eicosamoids: biosynthesis and interactions with immune response]. Medical Immunology (Russia) 2013; 15 (2): 119−130 (in Russian). https://doi.org/10.15789/1563-0625-2013-2-119-130

13. Augimeri G., Plastina P., Gionfriddo G., Rovito D., Giordano C., Fazio A., Barone I., Catalano S., Andò S., Bonofiglio D., Meijerink J, Witkamp R. N-Eicosapentaenoyl Dopamine, A Conjugate of Dopamine and Eicosapentaenoic Acid (EPA), Exerts Anti-inflammatory Properties in Mouse and Human Macrophages // Nutrients. 2019. Vol. 11, Iss. 9. Article number:2247 https://doi.org/10.3390/nu11092247