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Nutritional Immunology

Lutein and Eicosapentaenoic Acid Interact to Modify iNOS mRNA Levels through the PPAR/RXR Pathway in Chickens and HD11 Cell Lines¹

Department of Animal Science, University of California, Davis, CA 95616

² To whom correspondence should be addressed. Email: kcklasing{at}ucdavis.edu.

Two experiments were conducted to investigate the effect of lutein and fat or eicosapentaenoic acid (EPA) interaction on inducible nitric oxide synthase (iNOS), PPARs , ß, and , and retinoic acid X receptor (RXR) and mRNA levels. In Expt. 1, macrophages were collected from broiler chicks fed 3 or 6% dietary fat (g/100 g) with 0, 25, and 50 mg lutein/kg feed for 23 d. In Expt. 2, using a 3 x 3 factorial, eicosapentaenoic acid (EPA) at 0, 15 and 50µmol/L and lutein at 0, 10 and 100µmol/L were applied to HD11 cell culture for 24 h. In both experiments, cells were stimulated with lipopolysaccharide before RNA isolation. Lutein interacted with fat in Expt. 1 and with EPA in Expt. 2 to affect mRNA levels of iNOS, PPAR, and RXR in chicken macrophages and HD11 cells, respectively (P < 0.05). At 3% dietary fat or up to 15 µmol/L EPA in the medium, increasing lutein increased the iNOS mRNA. However, at 6% dietary fat or 50 µmol/L EPA, lutein did not cause a rise in iNOS mRNA. Increasing lutein in the medium from 0 to 100 µmol/L decreased iNOS mRNA. Increasing lutein with high fat (6%) or EPA (15 µmol/L EPA) increased PPAR and RXR mRNA levels. Lutein increased PPAR mRNA levels in both macrophages (P < 0.01) and HD11 (P = 0.01) cells and RXR (P < 0.01) mRNA levels in macrophages. GW9662, a PPAR antagonist, prevented (P < 0.01) the lutein-induced iNOS mRNA downregulation in HD11 cells. LG101208, a RXR antagonist, prevented (P < 0.01) iNOS upregulation induced by 10 µmol/L lutein and iNOS mRNA downregulation induced by 100 µmol/L lutein. We conclude that lutein and EPA interact through the PPAR and RXR pathways to modulate iNOS mRNA.

KEY WORDS: • lutein • fat • eicosapentaenoic acid • nuclear receptors

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