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Vol. 53, Issue 4, 663-669, April 1998
B
Departments of
Pharmacology (Z.N., Y.M., M.F., V.R.),
Surgery
(L.R.), and
Microbiology/Immunology (A.M.), Southern Illinois
University School of Medicine, Springfield, Illinois 62794 and
Department of Medicine (Cardiology) and Pharmacology (H.R., G.S.), Duke
University Medical Center, Durham, North Carolina 27710
The A1 adenosine receptor (A1AR) contributes to
the cytoprotective action of adenosine under conditions known to
generate reactive oxygen species (ROS). Pharmacological manipulation of A1AR expression has been shown to modulate this
cytoprotective role. In this study, we provide evidence that ROS
generated could increase the expression of the A1AR and
thereby offset the detrimental effects of ROS. Incubation of
DDT1MF-2 smooth muscle cells with ROS-generating
chemotherapeutic agents, such as cisplatin (2.5 µM) or
H2O2 (10 µM), elicited an
increase in A1AR expression within 24 hr. The induction by
H2O2 was reduced by the ROS scavenger catalase
but not superoxide dismutase. Inhibition of nuclear factor 
B
(NFB) by pyrrolidine dithiocarbamate (200 µM),
dexamethasone (100 nM), or genistein (1 µM)
abrogated the cisplatin-mediated increase in A1AR.
Cisplatin promoted rapid translocation of NFB (but not AP-1) to the
nucleus, as detected by electrophoretic mobility shift assays and by
Western blotting. A putative NFB sequence in the A1AR
promoter effectively competed with labeled B probe for binding in
nuclear preparations derived from DDT1MF-2 cells. Transient
transfection of DDT1MF-2 cells with the A1AR promoter coupled to firefly luciferase reporter gene led to
cisplatin-inducible and pyrrolidine dithiocarbamate-sensitive
luciferase activity, suggesting the presence of functional NFB
binding site(s) in the A1AR promoter sequence. Treatment of
cells with (R)-phenylisopropyladenosine (1 µM), an agonist of the A1AR, reduced
cisplatin-mediated lipid peroxidation, which was reversed after
blockade of the A1AR. These data suggest that ROS can
increase the expression of the A1AR by activating NFB
regulatory site(s) on this gene and thereby enhance the cytoprotective
role of adenosine.
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