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Division of Developmental Genetics, Center for Biomedical Science, Chiba University School of Medicine, Chiba, Japan
Activated macrophages produce nitric oxide (NO) that is an
important effector molecule for their antimicrobial and antitumor
activities. Since this NO is also toxic for themselves, they have
self-defense mechanisms. To elucidate the mechanisms in a physiologic
condition, expression of bcl-2 family genes were examined
in peritoneal macrophages and RAW264 macrophage cell line activated
with IFN-
and LPS. Bcl-xL, but not bcl-2 and
bax mRNA, was highly inducible within 3 h after
stimulation. The induction required new protein synthesis, but was
independent of effects of synthesized NO. Since activated RAW264 were
more resistant to NO-induced apoptosis mediated by the exposure to
S-nitroso-N-acetyl-penicillamine (SNAP) than
nonactivated RAW264, the inducible Bcl-xL may play a role
in the protection from NO toxicity. To confirm the protective function,
RAW264 were stably transfected with bcl-xL.
Those transfectants activated with IFN- and LPS appeared highly
resistant to NO-induced cell death detected within 24 h after
stimulation, although their NO production was similar to those of
parental RAW264 and neomycin control-transfected cells. Furthermore,
bcl-xL transfectants displayed substantial
protection from SNAP-induced apoptosis. These results establish a link
between self-defense to the synthesized NO and the induction of
Bcl-xL in activated macrophages.
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