(Hypertension. 2000;35:942.)
© 2000 American Heart Association, Inc.
Scientific Contributions |
Role of Reactive Oxygen Species in Bradykinin-Induced Mitogen-Activated Protein Kinase and c-fos Induction in Vascular Cells
From the Departments of Medicine and Cell and Molecular Pharmacology and Experimental Therapeutics, Medical University of South Carolina, Charleston.
Correspondence to Ayad A. Jaffa, PhD, Department of Medicine, Endocrinology-Diabetes-Medical Genetics, Medical University of South Carolina, 171 Ashley Ave, Charleston, SC 29425. E-mail jaffaa{at}musc.edu
Abstract—Bradykinin
stimulates proliferation of aortic vascular smooth muscle cells
(VSMCs). We investigated the action of bradykinin on the
phosphorylation state of the mitogen-activated
protein kinases p42mapk and p44mapk in VSMCs
and tested the hypothesis that reactive oxygen species (ROS) might be
involved in the signal transduction pathway linking bradykinin
activation of nuclear transcription factors to the
phosphorylation of p42mapk and
p44mapk. Bradykinin (10-8 mol/L) rapidly
increased (4- to 5-fold) the phosphorylation of
p42mapk and p44mapk in VSMCs. Preincubation of
VSMCs with either N-acetyl-L-cysteine and/or
-lipoic acid significantly decreased bradykinin-induced cytosolic
and nuclear phosphorylation of p42mapk and
p44mapk. In addition, the induction c-fos
mRNA levels by bradykinin was completely abolished by
N-acetyl-L-cysteine and -lipoic acid.
Using the cell-permeable fluorescent dye
dichlorofluorescein diacetate, we determined that
bradykinin (10-8 mol/L) rapidly increased the generation
of ROS in VSMCs. The NADPH oxidase inhibitor diphenylene
iodonium (DPI) blocked bradykinin-induced c-fos mRNA
expression and p42mapk and p44mapk activation,
implicating NADPH oxidase as the source for the generation of ROS.
These findings demonstrate that the phosphorylation of
cytosolic and nuclear p42mapk and p44mapk and
the expression of c-fos mRNA in VSMCs in response to
bradykinin are mediated via the generation of ROS and implicate ROS as
important mediators in the signal transduction pathway through which
bradykinin promotes VSMC proliferation in states of vascular
injury.
Key Words: bradykinin • kinases • proto-oncogenes • reactive oxygen species
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