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-induced endothelial cell apoptosis: dual regulation by reactive oxygen species
Division of Cardiology, Department of Medicine, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA
1Correspondence: Division of Cardiology, Department of Medicine, The Johns Hopkins University School of Medicine, Ross 1023, 720 Rutland Ave., Baltimore MD 21205, USA. E-mail: kirani{at}mail.jhmi.edu
Reactive oxygen species (ROS) have been implicated as mediators of tumor
necrosis factor-alpha (TNF) -induced apoptosis. In addition to leading
to cell death, ROS can also promote cell growth and/or survival. We
investigated these two roles of ROS in TNF-induced endothelial cell
apoptosis. Human umbilical vein endothelial cells (HUVECs) stimulated
with TNF produced an intracellular burst of ROS. Adenoviral-mediated
gene transfer of a dominant negative form of the small GTPase Rac1
(Rac1N17) partially suppressed the TNF-induced oxidative burst without
affecting TNF-induced mitochondrial ROS production. HUVECs were
protected from TNF-induced apoptosis. Expression of Rac1N17 blocked
TNF-induced activation of nuclear factor-kappa B (NF-
B), increased
activity of caspase-3, and markedly augmented endothelial cell
susceptibility to TNF-induced apoptosis. Direct inhibition of NF-B
through adenoviral expression of the super repressor form of inhibitor
of B
(I-B S32/36A) also increased susceptibility of HUVECs to
TNF-induced apoptosis. Rotenone, a mitochondrial electron transport
chain inhibitor, suppressed TNF-induced mitochondrial ROS production,
proteolytic cleavage of procaspase-3, and apoptosis. These findings
show that Rac1 is an important regulator of TNF-induced ROS production
in endothelial cells. Moreover, they suggest that Rac1-dependent ROS,
directly or indirecly, lead to protection against TNF-induced death,
whereas mitochondrial-derived ROS promote TNF-induced
apoptosis.—Deshpande, S. S., Angkeow, P., Huang, J., Ozaki,
M., Irani, K. Rac1 inhibits TNF--induced endothelial cell apoptosis:
dual regulation by reactive oxygen species.
Key Words: Apoptosis • ROS • Rac1 • nuclear factor-kappa B • caspase-3
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