Docosahexaenoic acid decreases IRF-1 MRNA and thus inhibits activation of both the IRF-E and NFκd response elements of the iNOS promoter

Mariusz L. Kielar, D. Rohan Jeyarajah, Jeffery G. Penfield, Christopher Y. Lu

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Abstract

Background. Nitric oxide produced by inducible nitric oxide synthase (iNOS) may be cytotoxic during cardiac, hepatic, and renal acute allograft rejection. Because the incidence of rejection is decreased by fish oils that contain docosahexaenoic acid (DHA), we investigated the effects of DHA on iNOS. Using nuclear run-on assays and iNOS-promoter constructs, we previously showed that docosahexaenoic acid (DHA) inhibits activation of the iNOS gene by murine macrophages that had been stimulated in vitro by IFNγ plus lipopolysaccharide. Methods. In our current investigation, our purpose has been to determine how DHA inhibits iNOS gene activation in murine macrophages, by using gel retardation and Northern blotting techniques. We studied the effects of DHA on the formation nuclear protein complexes that interact with the critical iNOS promoter's response elements for IRF-1 (IRF- E-923 to -913 bp) and NF-κB (NFκd -85 to -75 bp). Results. We now show that DHA inhibited increases of IRF-1 mRNA abundance in response to IFNy plus lipopolysaccharide. As expected, we found that this prevented formation of the nuclear protein complex that hinds to the IRF-E DNA response element. We also found that inhibition of IRF-1 inhibited formation of the nuclear protein complex that binds to the NFkd DNA response element. Conclusions. DHA decreases the abundance of IRF-1 mRNA in stimulated cells. That, in turn, results in the decreased nuclear protein binding to the major iNOS promoter response elements (IRF-E and NF-κB). We found that this occurred because IRF-1 is a component of both the nuclear protein complex that binds to IRF-E and the nuclear protein complex that binds to NFκd.

Original languageEnglish (US)
Pages (from-to)2131-2137
Number of pages7
JournalTransplantation
Volume69
Issue number10
StatePublished - May 27 2000

Fingerprint

Docosahexaenoic Acids
Response Elements
Nitric Oxide Synthase Type II
Nuclear Proteins
Lipopolysaccharides
Macrophages
Messenger RNA
Fish Oils
DNA
Protein Binding
Northern Blotting
Transcriptional Activation
Allografts
Nitric Oxide
Gels
Kidney
Liver
Incidence
Genes

ASJC Scopus subject areas

  • Transplantation
  • Immunology

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Docosahexaenoic acid decreases IRF-1 MRNA and thus inhibits activation of both the IRF-E and NFκd response elements of the iNOS promoter. / Kielar, Mariusz L.; Jeyarajah, D. Rohan; Penfield, Jeffery G.; Lu, Christopher Y.

In: Transplantation, Vol. 69, No. 10, 27.05.2000, p. 2131-2137.

Research output: Contribution to journalArticle

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abstract = "Background. Nitric oxide produced by inducible nitric oxide synthase (iNOS) may be cytotoxic during cardiac, hepatic, and renal acute allograft rejection. Because the incidence of rejection is decreased by fish oils that contain docosahexaenoic acid (DHA), we investigated the effects of DHA on iNOS. Using nuclear run-on assays and iNOS-promoter constructs, we previously showed that docosahexaenoic acid (DHA) inhibits activation of the iNOS gene by murine macrophages that had been stimulated in vitro by IFNγ plus lipopolysaccharide. Methods. In our current investigation, our purpose has been to determine how DHA inhibits iNOS gene activation in murine macrophages, by using gel retardation and Northern blotting techniques. We studied the effects of DHA on the formation nuclear protein complexes that interact with the critical iNOS promoter's response elements for IRF-1 (IRF- E-923 to -913 bp) and NF-κB (NFκd -85 to -75 bp). Results. We now show that DHA inhibited increases of IRF-1 mRNA abundance in response to IFNy plus lipopolysaccharide. As expected, we found that this prevented formation of the nuclear protein complex that hinds to the IRF-E DNA response element. We also found that inhibition of IRF-1 inhibited formation of the nuclear protein complex that binds to the NFkd DNA response element. Conclusions. DHA decreases the abundance of IRF-1 mRNA in stimulated cells. That, in turn, results in the decreased nuclear protein binding to the major iNOS promoter response elements (IRF-E and NF-κB). We found that this occurred because IRF-1 is a component of both the nuclear protein complex that binds to IRF-E and the nuclear protein complex that binds to NFκd.",
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AU - Lu, Christopher Y.

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N2 - Background. Nitric oxide produced by inducible nitric oxide synthase (iNOS) may be cytotoxic during cardiac, hepatic, and renal acute allograft rejection. Because the incidence of rejection is decreased by fish oils that contain docosahexaenoic acid (DHA), we investigated the effects of DHA on iNOS. Using nuclear run-on assays and iNOS-promoter constructs, we previously showed that docosahexaenoic acid (DHA) inhibits activation of the iNOS gene by murine macrophages that had been stimulated in vitro by IFNγ plus lipopolysaccharide. Methods. In our current investigation, our purpose has been to determine how DHA inhibits iNOS gene activation in murine macrophages, by using gel retardation and Northern blotting techniques. We studied the effects of DHA on the formation nuclear protein complexes that interact with the critical iNOS promoter's response elements for IRF-1 (IRF- E-923 to -913 bp) and NF-κB (NFκd -85 to -75 bp). Results. We now show that DHA inhibited increases of IRF-1 mRNA abundance in response to IFNy plus lipopolysaccharide. As expected, we found that this prevented formation of the nuclear protein complex that hinds to the IRF-E DNA response element. We also found that inhibition of IRF-1 inhibited formation of the nuclear protein complex that binds to the NFkd DNA response element. Conclusions. DHA decreases the abundance of IRF-1 mRNA in stimulated cells. That, in turn, results in the decreased nuclear protein binding to the major iNOS promoter response elements (IRF-E and NF-κB). We found that this occurred because IRF-1 is a component of both the nuclear protein complex that binds to IRF-E and the nuclear protein complex that binds to NFκd.

AB - Background. Nitric oxide produced by inducible nitric oxide synthase (iNOS) may be cytotoxic during cardiac, hepatic, and renal acute allograft rejection. Because the incidence of rejection is decreased by fish oils that contain docosahexaenoic acid (DHA), we investigated the effects of DHA on iNOS. Using nuclear run-on assays and iNOS-promoter constructs, we previously showed that docosahexaenoic acid (DHA) inhibits activation of the iNOS gene by murine macrophages that had been stimulated in vitro by IFNγ plus lipopolysaccharide. Methods. In our current investigation, our purpose has been to determine how DHA inhibits iNOS gene activation in murine macrophages, by using gel retardation and Northern blotting techniques. We studied the effects of DHA on the formation nuclear protein complexes that interact with the critical iNOS promoter's response elements for IRF-1 (IRF- E-923 to -913 bp) and NF-κB (NFκd -85 to -75 bp). Results. We now show that DHA inhibited increases of IRF-1 mRNA abundance in response to IFNy plus lipopolysaccharide. As expected, we found that this prevented formation of the nuclear protein complex that hinds to the IRF-E DNA response element. We also found that inhibition of IRF-1 inhibited formation of the nuclear protein complex that binds to the NFkd DNA response element. Conclusions. DHA decreases the abundance of IRF-1 mRNA in stimulated cells. That, in turn, results in the decreased nuclear protein binding to the major iNOS promoter response elements (IRF-E and NF-κB). We found that this occurred because IRF-1 is a component of both the nuclear protein complex that binds to IRF-E and the nuclear protein complex that binds to NFκd.

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