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(Received for publication, November 20, 1996, and in revised form, February 13, 1997)
From the Department of The understanding of the induction and regulation
of inducible nitric-oxide synthase (iNOS) in human cells may be
important in developing therapeutic interventions for inflammatory
diseases. In the present study, we not only demonstrated that human
fetal mixed glial cultures, as well as enriched microglial cultures, synthesize iNOS and nitric oxide (NO) in response to cytokine stimulation, but also assessed the kinetics of iNOS and NO synthesis in
human fetal mixed glial cultures. The iNOS mRNA was expressed within 2 h after stimulation and decreased to base line by 2 days. Significant levels of iNOS protein appeared within 24 h after stimulation and remained elevated during the culture period. A dramatic
increase in NO production and NO-mediated events, such as the induction
of cyclic guanosine monophosphate (cGMP), NADPH diaphorase activity,
and nitrotyrosine occurred 3 days after stimulation, a delay of 48 h from the time of the first expression of iNOS enzyme. This delay of
NO production was altered by the addition of tetrahydrobiopterin, but
not by the addition of L-arginine, heme, flavin
adenine dinucleotide (FAD), flavin mononucleotide (FMN), or NADPH.
These findings suggest that a post-translational regulatory event might
be involved in iNOS-mediated NO production in human glia.
Volume 272, Number 17,
Issue of April 25, 1997
pp. 11327-11335
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
A KINETIC ANALYSIS
,
,
,
,
and
Neurology and 
Medical
and Molecular Pharmacology, UCLA School of Medicine, Los Angeles,
California 90095 and the ¶ Department of Immunology, Berlex
Biosciences, Richmond, California 94804
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