Original ContributionNox5 mediates PDGF-induced proliferation in human aortic smooth muscle cells
Section snippets
Materials
PDGF-BB was purchased from R&D Systems, Inc. (Minneapolis, MN), and N-acetylcysteine (NAC) was from Sigma-Aldrich (St. Louis, MO). Non-silencing siRNA and siRNA to Nox5 (siNox5, which targets all Nox5 isoforms) were purchased from Ambion (Austin, TX). Phospho-JAK2 (Tyr1007/Tyr1008) antibody was from Santa Cruz Biotechnologies (Santa Cruz, CA), phospho-STAT3 (Tyr705) and STAT3 antibodies were from Cell Signaling Technology, Inc (Danvers, MA), and anti-JAK2 antibody was from Millipore (Billerica,
HASMCs predominantly express Nox4 and all isoforms of Nox5
Real time PCR with primers to Nox family members revealed that proliferating HASMCs express Nox4 and Nox5, with Nox4 mRNA more abundant than Nox5 mRNA (23,895 ± 7,150 vs. 395 ± 56 copies per 108 18S). Using primers to Nox1 and Nox2, we were able to measure copy numbers as low as 10 per reaction, but detected minimal Nox1 and no Nox2 mRNA in HASMCs. Five variants of Nox5 have been reported: α, β, γ, δ, and a short form that is missing exon 6 [24], [31], [32]. We used reverse transcriptase PCR using
Discussion
The NADPH oxidases have emerged as important elements of signaling pathways in VSMCs. There is ample evidence that more than one Nox exists in this cell type and that the different homologues are located in different compartments [34] and serve different functions. In rat VSMCs, Nox1 appears to be required for angiotensin II- and PDGF-induced ROS production and growth [22], and Nox4 has been shown to be important for the maintenance of a contractile, non-proliferative phenotype [23]. However,
Acknowledgments
This work was supported by NIH grant HL074604 to DBJ and HL058863 to KKG. We thank Dr. Peter Sayeski for his advice on the JAK/STAT experiments.
References (37)
- et al.
Induction of B-type receptors for platelet-derived growth factor in vascular inflammation: possible implications for development of vascular proliferative lesions
Lancet
(1988) PDGF and cardiovascular disease
Cytokine Growth Factor Rev.
(2004)- et al.
A Ca(2+)-activated NADPH oxidase in testis, spleen, and lymph nodes
J. Biol. Chem.
(2001) - et al.
cAMP-response element-binding protein mediates acid-induced NAD PH oxidase NOX5-S expression in Barrett esophageal adenocarcinoma cells
J. Biol. Chem.
(2006) - et al.
NOX5 variants are functionally active in endothelial cells
Free Radic. Biol. Med.
(2007) - et al.
Mechanism of Ca2+ activation of the NADPH oxidase 5 (NOX5)
J. Biol. Chem.
(2004) - et al.
Homologs of gp91phox: cloning and tissue expression of Nox3, Nox4, and Nox5
Gene
(2001) - et al.
Synchronization of cultured vascular smooth muscle cells following reversal of quiescence induced by treatment with the antioxidant N-acetylcysteine
Exp. Cell Res.
(1998) - et al.
Novel mechanism of activation of NADPH oxidase 5. calcium sensitization via phosphorylation
J. Biol. Chem.
(2007) - et al.
NADPH oxidase 5 (NOX5) interacts with and is regulated by calmodulin
FEBS Lett.
(2007)
Human monocyte-endothelial cell interaction induces platelet-derived growth factor expression
Cardiovasc. Res.
Local expression of inflammatory cytokines in human atherosclerotic plaques
J. Atheroscler. Thromb.
A platelet-dependent serum factor that stimulates the proliferation of arterial smooth muscle cells in vitro
Proc. Natl. Acad. Sci. U.S.A.
Immunohistochemical localization of PDGF B, PDGF beta receptors and IGF I receptors during atherogenesis
Zentralblatt fur Pathologie
Endogenous neutralizing antibodies against platelet-derived growth factor-AA inhibit atherogenesis in the cholesterol-fed rabbit
Arterioscler. Thromb. Vasc. Biol.
Endogenously elicited antibodies to platelet derived growth factor-BB and platelet cytosolic protein inhibit aortic lesion development in the cholesterol-fed rabbit
Int. J. Exp. Pathol.
Inhibition of neointimal smooth muscle accumulation after angioplasty by an antibody to PDGF
Science
Bolus endovascular PDGFR-beta antisense treatment suppressed intimal hyperplasia in a rat carotid injury model
Circulation
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