Dietary arachidonate in milk replacer triggers dual benefits of PGE signaling in LPS-challenged piglet alveolar macrophages.

BACKGROUND

Respiratory infections challenge the swine industry, despite common medicinal practices. The dual signaling nature of PGE (supporting both inflammation and resolution) makes it a potent regulator of immune cell function. Therefore, the use of dietary long chain n-6 PUFA to enhance PGE effects merits investigation.

METHODS

Day-old pigs ( = 60) were allotted to one of three dietary groups for 21 d ( = 20/diet), and received either a control diet (CON, arachidonate = 0.5% of total fatty acids), an arachidonate (ARA)-enriched diet (LC n-6, ARA = 2.2%), or an eicosapentaenoic (EPA)-enriched diet (LC n-3, EPA = 3.0%). Alveolar macrophages and lung parenchymal tissue were collected for fatty acid analysis. Isolated alveolar macrophages were stimulated with LPS in situ for 24 h, and mRNA was isolated to assess markers associated with inflammation and eicosanoid production. Culture media were collected to assess PGE secretion. Oxidative burst in macrophages was measured by: 1) oxygen consumption and extracellular acidification (via Seahorse), 2) cytoplasmic oxidation and 3) nitric oxide production following 4, 18, and 24 h of LPS stimulation.

RESULTS

Concentration of ARA (% of fatty acids, /) in macrophages from pigs fed LC n-6 was 86% higher than CON and 18% lower in pigs fed LC n-3 ( < 0.01). Following LPS stimulation, abundance of and mRNA ( <  0.0001), and PGE secretion ( < 0. 01) were higher in LC n-6 PAM vs. CON. However, abundance was 1.6-fold lower than CON. Macrophages from CON and LC n-6 groups were 4-fold higher in abundance ( < 0.0001) compared to LC n-3. Oxygen consumption and extracellular acidification rates increased over 4 h following LPS stimulation ( < 0.05) regardless of treatment. Similarly, increases in cytoplasmic oxidation ( < 0.001) and nitric oxide production ( <  0.002) were observed after 18 h of LPS stimulation but were unaffected by diet.

CONCLUSIONS

We infer that enriching diets with arachidonic acid may be an effective means to enhance a stronger innate immunologic response to respiratory challenges in neonatal pigs. However, further work is needed to examine long-term safety, clinical efficacy and economic viability.