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J Biol Chem, Vol. 273, Issue 20, 12219-12226, May 15, 1998
From the Department of Pediatrics, Medical University of South
Carolina, Charleston, South Carolina 29425
Nitric oxide produced by inducible nitric-oxide
synthase (iNOS) in different cells including brain cells in response to
proinflammatory cytokines plays an important role in the
pathophysiology of stroke and other neurodegenerative diseases. The
present study underlines the importance of protein phosphatase (PP) 1 and 2A in the regulation of the differential expression of iNOS in rat
primary astrocytes and macrophages. Compounds (calyculin A,
microcystin, okadaic acid, and cantharidin) that inhibit PP 1 and 2A
were found to stimulate the lipopolysaccharide (LPS)- and
cytokine-mediated expression of iNOS and production of NO in rat
primary astrocytes and C6 glial cells. However, these
inhibitors inhibited the LPS- and cytokine-mediated expression of iNOS
and production of NO in rat resident macrophages and RAW 264.7 cells.
Similarly, okadaic acid, an inhibitor of PP 1/2A, stimulated the iNOS
promoter-derived chloramphenicol acetyltransferase activity in
astrocytes and inhibited the iNOS promoter-derived chloramphenicol
acetyltransferase activity in macrophages, indicating that okadaic acid
also differentially regulates the transcription of the iNOS gene in
astrocytes and macrophages. The observed stimulation of the expression
of iNOS in astrocytes and the inhibition of the expression of iNOS in macrophages with the inhibition of PP 1/2A activity clearly delineate a
novel role of PP 1/2A in the differential regulation of iNOS in rat
astrocytes and macrophages. Because the activation of NF-
Inhibitors of Protein Phosphatase 1 and 2A Differentially
Regulate the Expression of Inducible Nitric-oxide Synthase in Rat
Astrocytes and Macrophages
B is
necessary for the induction of iNOS and the expression of tumor necrosis factor (TNF)-
also depends on the activation of NF-B, we
examined the effect of okadaic acid on the LPS-mediated activation of
NF-B and production of TNF- in rat primary astrocytes and macrophages. Interestingly, in both cell types, okadaic acid stimulated the LPS-mediated DNA binding as well as transcriptional activity of
NF-B and production of TNF-. This study suggests that the stimulation of iNOS expression in astrocytes by inhibitors of PP 1/2A
is possibly due to the stimulation of NF-B activation; however,
activation of NF-B is not sufficient for the induction of iNOS in
macrophages and that apart from NF-B some other signaling pathway(s)
sensitive to PP 1 and/or PP 2A is/are possibly involved in the
regulation of iNOS in macrophages. This differential induction of iNOS
as compared with similar activation of NF-B by inhibitors of PP 1/2A
indicates the involvement of different intracellular signaling events
for the induction of iNOS in two cell types of the same animal
species.
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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