Docosahexaenoic acid and TUG-891 activate free fatty acid-4 receptor in bovine neutrophils.

Fatty acids are well known metabolic intermediaries but also have a role in the immune response. Long-chain fatty acids such as omega-6 and -9 activate neutrophil function through free fatty acid (FFA)-1 receptor in bovines. Although omega-3 has also been suggested to influence neutrophil function, the details remain unclear. The goal of this study was to determine the presence of the bovine FFA4 receptor and its effect on neutrophil responses. We treated bovine neutrophils with the natural and synthetic agonists of FFA4 receptor docosahexaenoic acid (DHA) and TUG-891, respectively, and assessed oxidative and no oxidative response. We detected protein and mRNA FFA4 receptor expression through immunofluorescence, immunoblot, and RT-PCR analysis. DHA and TUG-891 both increased intracellular calcium mobilisation in bovine neutrophils, with 50% effective concentrations of 99 μM and 73 μM, respectively, which was partially reduced after treatment with the FFA4 antagonist AH7614. Furthermore, DHA and TUG-891 increased matrix metalloproteinase (MMP)-9 granules release and superoxide production. AH7614 and the intracellular calcium chelator BAPTA-AM decreased the superoxide production induced by TUG-891 and by both DHA and TUG-891, respectively, suggesting a key role of intracellular calcium in FFA4 agonists-induced superoxide production. These results highlight an important mechanism of bovine neutrophil responses mediated via FFA4 receptor, which can further inform the development of new formulations for DHA-enriched feed supplements to enhance innate immunity in dairy cattle.