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Vol. 55, Issue 3, 481-488, March 1999
Institute of Pharmacology, College of Medicine, National Taiwan
University, Taipei, Taiwan
Protein kinase C (PKC)-
, -
I, and -
are known to be
involved in the lipopolysaccharide (LPS)-induced nitric oxide (NO)
production in RAW 264.7 macrophages. The role of mitogen-activated
protein kinases (MAPK) p44/42 and p38 in the LPS effect was studied
further. LPS-mediated NO release and the inducible form of NO synthase expression were inhibited by the p38 inhibitor, SB 203580, but not by
the MAPK kinase inhibitor, PD 98059. Ten-minute treatment of cells with
LPS resulted in the activation of p44/42 MAPK, p38, and c-Jun
NH2-terminal kinase. Marked or slight activation,
respectively, of p44/42 MAPK or p38 was also seen after 10-min
treatment with 12-O-tetradecanoylphorbol-13-acetate, but
c-Jun NH2-terminal kinase activation did not occur.
Tyrosine kinase inhibitor, genestein, attenuated the LPS-induced
activation of both p44/42 MAPK and p38, whereas the PKC inhibitors, Ro
31-8220 and calphostin C, or long-term treatment with
12-O-tetradecanoylphorbol-13-acetate resulted in
inhibition of p44/42 MAPK activation, but had only a slight effect on
p38 activation, indicating that LPS-mediated PKC activation resulted in
the activation of p44/42 MAPK. Nuclear factor-
B (NF-B)-specific
DNA-protein-binding activity in the nuclear extracts was enhanced by
10-min, 1-h, or 24-h treatment with LPS. Analysis of the proteins
involved in NF-B binding showed translocation of p65 from the
cytosol to the nucleus after 10-min treatment with LPS. The onset of
NF-B activation correlated with the cytosolic degradation of both
inhibitory proteins of NF-B, IB- and IB-.
IB- was resynthesized rapidly after loss (1-h LPS treatment),
whereas IB- levels were not restored until after 24-h treatment.
SB 203580 but not PD 98059 inhibited the LPS-induced stimulation of
NF-B DNA-protein binding. Thus, activation of p38 but not p44/42
MAPK by LPS resulted in the stimulation of NF-B-specific DNA-protein
binding and the subsequent expression of inducible form of NO synthase
and NO release in RAW 264.7 macrophages.
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