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*Compound via MeSH
*Substance via MeSH
Hazardous Substances DB
*CYCLOHEXIMIDE
*NITRIC OXIDE
The Journal of Immunology, 1999, 162: 4191-4197.
Copyright © 1999 by The American Association of Immunologists

Regulation of Cytosolic COX-2 and Prostaglandin E2 Production by Nitric Oxide in Activated Murine Macrophages

Rajesh Patel*, Mukundan G. Attur*, Mandar Dave*, Steven B. Abramson*,{dagger} and Ashok R. Amin1,*,{dagger},{ddagger}

* Department of Rheumatology, Hospital for Joint Diseases, New York, NY 10003; and Departments of {dagger} Medicine and {ddagger} Pathology, Kaplan Cancer Center, New York University Medical Center, New York, NY 10016.

Murine macrophages (RAW 264.7) when stimulated with LPS show 90% distribution of cyclooxygenase-2 (COX-2) in the nuclear fraction and ~10% in the cytosolic fraction. Further analysis of this cytosolic fraction at 100,000 x g indicates that the COX-2 is distributed both in the 100,000 x g soluble fraction and membrane fraction. Stimulation of RAW 264.7 cells with LPS in the presence of inducible nitric oxide synthase inhibitor L-NMMA at concentrations that inhibit nitrite accumulation by <=80% is inadequate to augment PGE2 production. However, inhibition of nitrite accumulation by >=85% with higher concentrations of L-NMMA shows 1) up-regulation of PGE2 production, 2) accumulation of COX-2 protein in the 100,000 x g soluble and membrane fractions of the cytosolic fraction, and 3) with no significant effects on the accumulation of COX-2 mRNA. These experiments suggest that low concentrations of nitric oxide (10–15% of the total) attenuate PGE2 production in response to LPS in RAW 264.7 cells. This inhibition is, in part, due to decreased expression of cytosolic COX-2 protein.




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