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作者（2019）在《Dietary arachidonate in milk replacer triggers dual benefits of PGE2 signaling in LPS-challenged piglet alveolar macrophages》一文中研究指出：Background: Respiratory infections challenge the swine industry, despite common medicinal practices. The dual signaling nature of PGE2（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 PGE2 effects merits investigation.Methods: Day-old pigs（n = 60） were allotted to one of three dietary groups for 21 d（n = 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 m RNA was isolated to assess markers associated with inflammation and eicosanoid production.Culture media were collected to assess PGE2 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, w/w） in macrophages from pigs fed LC n-6 was 86% higher than CON and 18% lower in pigs fed LC n-3（P < 0.01）. Following LPS stimulation, abundance of COX-2 and TNF-α mRNA（P < 0.0001）, and PGE2 secretion（P < 0. 01） were higher in LC n-6 PAM vs. CON. However, ALOX5 abundance was1.6-fold lower than CON. Macrophages from CON and LC n-6 groups were 4-fold higher in ALOX12/15 abundance（P < 0.0001） compared to LC n-3. Oxygen consumption and extracellular acidification rates increased over 4 h following LPS stimulation（P < 0.05） regardless of treatment. Similarly, increases in cytoplasmic oxidation（P < 0.001）and nitric oxide production（P < 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.
Abstract
Background: Respiratory infections challenge the swine industry, despite common medicinal practices. The dual signaling nature of PGE2（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 PGE2 effects merits investigation.Methods: Day-old pigs（n = 60） were allotted to one of three dietary groups for 21 d（n = 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 m RNA was isolated to assess markers associated with inflammation and eicosanoid production.Culture media were collected to assess PGE2 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, w/w） in macrophages from pigs fed LC n-6 was 86% higher than CON and 18% lower in pigs fed LC n-3（P < 0.01）. Following LPS stimulation, abundance of COX-2 and TNF-α mRNA（P < 0.0001）, and PGE2 secretion（P < 0. 01） were higher in LC n-6 PAM vs. CON. However, ALOX5 abundance was1.6-fold lower than CON. Macrophages from CON and LC n-6 groups were 4-fold higher in ALOX12/15 abundance（P < 0.0001） compared to LC n-3. Oxygen consumption and extracellular acidification rates increased over 4 h following LPS stimulation（P < 0.05） regardless of treatment. Similarly, increases in cytoplasmic oxidation（P < 0.001）and nitric oxide production（P < 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.
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