doi: 10.1161/hq0701.092135
© 2001 American Heart Association, Inc.
Vascular Biology |
4-Hydroxynonenal Prevents NO Production in Vascular Smooth Muscle Cells by Inhibiting Nuclear Factor-
B–Dependent Transcriptional Activation of Inducible NO Synthase
From the Department of Endocrinology and Metabolism, Dokkyo University School of Medicine, Mibu, Tochigi, Japan.
Correspondence to Dr Yoshiyuki Hattori, Department of Endocrinology and Metabolism, Dokkyo University School of Medicine, Mibu, Tochigi 321-0293, Japan. E-mail yhattori{at}dokkyomed.ac.jp
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Abstract |
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Abstract—The role of lipid peroxidation products in atherogenesis was studied. We investigated whether 4-hydroxy-2-nonenal (HNE) modulates activation of the nuclear factor (NF)-
B system or alters expression
of the NF-B target gene product, inducible NO synthase (iNOS),
in vascular smooth muscle cells (VSMCs) stimulated by
lipopolysaccharide (LPS) in combination with interferon
(IFN)-
(LPS/IFN). NO production induced by LPS/IFN was
dose-dependently inhibited by HNE. NF-B activation by LPS/IFN was
inhibited by HNE in a dose-dependent manner. HNE significantly
decreased LPS/IFN-stimulated proteolysis of IB-
. iNOS promoter
activity stimulated by LPS/IFN was also decreased by HNE
dose-dependently. The treatment of VSMCs with LPS/IFN strongly
stimulated iNOS mRNA and protein expression. The LPS/IFN-induced
increases in iNOS mRNA and protein levels were dose-dependently
decreased by HNE. Our data suggest that treatment with HNE blocks
signaling events required for IB- degradation, thereby preventing
NF-B activation. Inhibition of NF-B–regulated gene expression,
especially modulation of NO production, may contribute to
atherogenesis.
Key Words: 4-hydroxy-2-nonenal • nuclear factor-B • nitric oxide • vascular smooth muscle cells
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Introduction |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Atherosclerotic lesion formation is a complex process, which is in part mediated by inflammatory and oxidative mechanisms, including lipid peroxidation. Oxidized LDL has been shown to be present in atherosclerotic lesions.1 2 Oxidized LDL is a heterogeneous mixture of various components, including a variety of reactive aldehyde products that are formed during the oxidative degradation of polyunsaturated fatty acids.3 There is increasing evidence that aldehydes generated endogenously during the process of lipid peroxidation are causally involved in most of the pathophysiological effects associated with inflammatory and oxidative stress in cells and tissues.4 Lipid peroxidation–derived aldehydes appear not only to be end products and remnants of lipid peroxidation processes but also may act as mediators for the primary free radicals that initiate lipid peroxidation. Among the lipid peroxidation–derived aldehydes, 4-hydroxy-2-nonenal (HNE) is believed to be largely responsible for the cytopathological effects observed during inflammatory and oxidative processes.4
Nuclear factor (NF)-B, a pleiotropic transcription
factor, has been suggested to play an important role in gene regulation
during the inflammatory and immune reactions that promote
atherosclerotic
lesions.5 6 7
NF-B regulates the inducible expression of a variety of genes
involved in inflammatory and immune responses, including inducible NO
synthase
(iNOS).8 9 10 11
NO appears to play a key role in mediating vascular
remodeling.12 13 14
HNE-modified lysine residues have been identified in atherosclerotic
lesions,15 and a variety of
HNE effects in different cell types have been
described.16 17 18 19 20
HNE may modulate arterial wall NO production in
atherosclerotic lesions.
In the present study, we sought to define the effects of
HNE on iNOS expression in vascular smooth muscle cells (VSMCs). We
investigated whether HNE is able to modulate activation of the NF-B
system or alter the expression of the NF-B target gene product
iNOS. We examined to what extent this aldehyde affects B-dependent
and iNOS promoter–dependent transcriptional activity, the expression
of iNOS mRNA and protein, and NO
production.
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Methods |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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Cell Culture and RNA Extraction
Rat VSMCs were isolated by elastase and collagenase digestion of thoracic aortas from male Wistar rats, as previously described.21 Cells in passages 10 to 15 were used for experiments. Total RNA was extracted from confluent human VSMCs by using a modified guanidinium isothiocyanate method.22
Nitrite Assay
Nitrite accumulation, an indicator of NO synthesis,
was measured in the cell culture medium of confluent
VSMCs.23 Nitrite was
quantified colorimetrically after adding 100 µL of
Griess reagent (1% sulfanilamide and 0.1%
naphthylethylenediamine in 5% phosphoric acid) to 100-µL
samples. Absorbance at 550 nm was determined by using a microplate
reader (Molecular Devices). Nitrite
concentrations were calculated by comparison with the absorbance of
standard solutions of sodium nitrite prepared in cell culture
medium.
Cell Respiration
Cell respiration, an indicator of cell viability, was
assessed by the mitochondrial-dependent reduction of
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to
formazan.24 To examine the
cytotoxic effect of HNE, the cells were incubated (37°C) with MTT
(0.4 mg/mL) for a further 60 minutes after a 24-hour incubation for
nitrite assay. Culture medium was removed by aspiration, and the cells
were solubilized in dimethyl sulfoxide. The extent of reduction of MTT
to formazan within cells was quantified by the measurement of
optical density at 550 nm. Formazan production is
compared with the values obtained from the control cells (no
HNE).
iNOS mRNA Analysis
An iNOS cDNA was kindly provided by Dr Y.
Nunokawa24A (Suntory Co,
Osaka, Japan) and labeled with
[-32P]dCTP by random priming for use as
a Northern blot probe to assess iNOS mRNA expression. Northern blot
procedures were performed as previously
described.25 After probing
for iNOS expression, filters were stripped and reprobed for the
presence of GAPDH mRNA. The blots were exposed to an imaging plate
(Fuji Photo Film Co) at room temperature for 6 hours and
analyzed by using a FUJIX bioimaging analyzer
(BAS2000II, Fuji Photo Film Co).
NF-B Activation
To study NF-B activation, the cells were stably
transfected with a cis-reporter
plasmid containing a luciferase reporter gene linked to 5 repeats of
NF-B binding sites (pNFB-Luc,
Stratagene).25
For this, the pNFB-Luc plasmid was transfected together with a
pSV2neo helper plasmid (Clontech) into rat VSMCs
by using the FuGEN 6 transfection reagent
(Boehringer-Mannheim). The cells were cultured in the presence
of G418 (Clontech) at a concentration of 500
µg/mL with medium replacement at 2- to 3-day intervals. Approximately
3 weeks later, G418-resistant clones were isolated by using a
cloning cylinder and analyzed individually for expression of
luciferase activity. Several clones were selected for analysis
of NF-B activation. Luciferase activity was measured by using a
luciferase assay kit
(Stratagene).
iNOS Promoter Activity Assay
To study iNOS promoter function, VSMCs were stably
transfected with a construct containing a 1.7-kb fragment of the iNOS
promoter, which was cloned in front of a reporter gene encoding
secreted alkaline phosphatase (SEAP), as previously
described.26 27
SEAP activity, released into the cell culture medium, was measured by a
sensitive chemiluminescent assay (Phospha-Light,
TROPIX).
Western Blot of iNOS and IB-
After treatment, VSMC monolayers were lysed by using
an electrophoresis sample buffer. The protein concentration of the
samples was measured by use of a Bio-Rad
detergent-compatible protein assay. Subsequently, ß-mercaptoethanol
was added to the samples at a final concentration of 1%, and samples
were denatured by boiling for 5 minutes. Samples containing 10 µg of
protein were electrophoresed on 7% SDS-polyacrylamide gels and
transferred to PVD membranes (Bio-Rad), which
were incubated with a rabbit polyclonal antibody against human
IB- (1:2000, Santa Cruz Biotechnology) or a mouse monoclonal IgG1
antibody against mouse iNOS (1:2000, Transduction
Laboratories). IB- antibody or iNOS antibody binding
was detected by using donkey anti-rabbit or sheep anti-mouse IgG
horseradish peroxidase (1:20 000) and the ECL Plus system
(Amersham).
Statistical Analysis
Data are presented as mean±SEM of
measurements in 2 or 3 different VSMC cell cultures. Multiple
comparisons were evaluated by ANOVA followed by the Fisher protected
least significant difference test. The Student unpaired
t test was used for comparisons
between 2 experiments. A value of
P<0.05 was considered
statistically significant.
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Results |
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Our initial studies characterized NO production in VSMCs by determining the cumulative production of nitrite in culture supernatants at various intervals after treatment with lipopolysaccharide (LPS) in combination with interferon (IFN)-
(LPS/IFN). A lag phase of 6 to 8 hours preceded the induction
of nitrite synthesis, followed by a progressive increase in nitrite
synthesis for at least 24 hours.
Studies with different concentrations of HNE (5 to 100
µmol/L) showed a concentration-dependent inhibition of nitrite
production when HNE was added to VSMCs before LPS/IFN
(Figure 1a
). Although 50 µmol/L HNE produced an 
70% to
75% decline in nitrite production after LPS/IFN stimulation, 5
µmol/L HNE had little effect. Our cytotoxicity studies showed that
even at the highest concentration of HNE, only a marginal reduction in
cell viability was achieved (ie, there was little effect on MTT values
compared with the effect in the absence of HNE; inset to
Figure 1a).
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Time-course studies using a final concentration of 25
µmol/L HNE were conducted to determine whether HNE had a direct
affect on VSMC NO synthase (NOS) activity, ie, altered NOS expression
by VSMCs
(Figure 1b
). When HNE was added at increasing intervals after
the stimulation of VSMCs with LPS/IFN, the inhibition of nitrite
production decreased as the interval lengthened.
Next, experiments were performed to examine whether HNE
affects activation of the NF-B system. NF-B activation was
detected by measuring NF-B–dependent transcription in rat VSMCs
stably transfected with a luciferase reporter construct. Cells were
treated with different concentrations of HNE and then stimulated with
LPS/IFN for 3 hours. In the absence of HNE, we observed the expected
activation of NF-B by LPS/IFN. This increase was slightly affected
by HNE at 6.25 µmol/L, significantly decreased by 12.5 and 25
µmol/L, and completely abolished by 50 µmol/L. No effect of HNE
alone on NF-B activity was observed
(Figure 2). We then examined whether HNE could affect the
activation-induced proteolysis of the IB inhibitor
protein that traps the NF-B dimer in the cytosol. Incubation of
VSMCs with LPS/IFN over 1 hour led to a significant proteolysis of
IB-. HNE, when added before LPS/IFN, significantly decreased
LPS/IFN-stimulated proteolysis of IB-
(Figure 3A). Pretreatment with the proteosome
inhibitor MG115 also inhibited IB- degradation
(Figure 3B).
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We also tested whether HNE modulates activation of the iNOS
promoter. iNOS promoter function was determined by measuring iNOS
promoter–dependent transcription in rat VSMCs stably transfected with
a SEAP reporter construct containing the iNOS promoter. iNOS promoter
activity increased in the cells treated with LPS/IFN by 5-fold
relative to unstimulated levels. This increase was dose-dependently
decreased by HNE
(Figure 4).
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Next, we investigated whether the inhibition of
transcriptional activity by HNE has a corresponding impact on gene and
protein expression. For this purpose, we evaluated iNOS mRNA levels and
iNOS protein levels by Northern and Western blot analyses and
NO production by monitoring the accumulation of nitrite in the
cell supernatant. Although iNOS mRNA levels approached the detection
limit by Northern blot analysis in unstimulated VSMCs,
treatment with LPS/IFN provided a strong stimulus for iNOS mRNA
expression. The LPS/IFN-induced increase in iNOS mRNA level was
dose-dependently decreased by HNE
(Figure 5). As shown in
Figure 6, iNOS protein levels were very low in unstimulated
cells. Treatment with LPS/IFN caused a substantial induction of iNOS
protein expression in the cells. This induction of iNOS protein was
significantly decreased by HNE at 12.5 µmol/L and abolished at 50
µmol/L.
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Discussion |
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TopAbstractIntroductionMethodsResultsDiscussionReferences |
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The present report demonstrates that HNE, one of the most prominent aldehyde substances generated during extensive lipid peroxidation of oxidized LDL, inhibits the activation of NF-
B and
B-dependent transcription in VSMCs. Furthermore, iNOS
promoter–dependent transcription and the induction of iNOS mRNA and
protein were dose-dependently reduced in the presence of HNE, which
implies that the effect of HNE on the NF-B system is associated with
functional consequences. Indeed, HNE inhibited the induction of NO
production in VSMCs stimulated with LPS/IFN.
Our data show that HNE prevented the degradation of the
cytosolic NF-B inhibitor protein IB- after LPS/IFN
stimulation. Our cytotoxicity studies show that even at the highest
concentration of HNE, only a marginal reduction in cell viability was
achieved. IB proteolysis is mediated by the
proteosome,28 and our data
show that a proteosome inhibitor blocked degradation of
IB. Thus, the inhibitory effect of HNE on NF-B is not
due to general cytotoxicity, but HNE appears to selectively block
signaling events that are required for IB degradation, thereby
preventing NF-B activation. It has been reported that in cells of
the monocytic lineage, HNE specifically prevents LPS-induced
phosphorylation of IB-, whereas the profound
effect of HNE on NF-B is not due to interference with the
LPS-binding properties of the
cells.20 The authors suggest
that HNE appears to inhibit IB phosphorylation at a
signaling stage located downstream from the LPS receptor level. This is
also likely to be one of the mechanisms involved in HNE inhibition of
the NF-B system in VSMCs, and the detailed mechanism by which this
occurs, including HNE-induced IB degradation in stimulated VSMCs, is
currently under investigation.
Atherosclerosis is associated with reduced endothelium-dependent vasodilation, which has been ascribed to reduced NO formation.29 30 Constitutively expressed endothelial NOS is responsible for the regulated production of NO in arteries, whereas iNOS is expressed under inflammatory conditions.12 13 14 The loss of endothelial NOS may be compensated for by increased iNOS expression. As shown previously, HNE is present in atherosclerotic lesions.2 15 Because HNE can cause oxidative stress and inhibit iNOS, the antiatherogenic effects of NO and the ratio of NO to superoxide anion may be reduced in atherosclerotic arteries, leading to very rapid reaction of NO with superoxide anion and production of highly reactive peroxynitrite, which can directly damage lipids and protein.
It has been reported that HNE induced relaxation of human
cerebral arteries31 and
mesenteric arteries32 in an
endothelium-dependent manner. HNE could induce
intracellular peroxide
production33 in
endothelial cells, which may be a mediator of
endothelium-dependent
relaxation.34 It should be
evaluated whether HNE causes endothelial cell NOS
dysfunction, which could trigger the compensatory response (superoxide
anion generation followed by hydrogen peroxide production),
possibly representing an important mechanism underlying
oxidative vascular injury.34
HNE treatment of the cells resulted in depletion of intracellular
glutathione (GSH).33 GSH
plays an important role in the maintenance of the intracellular
redox balance and the regulation of several redox-sensitive
transcription factors, such as NF-B. Because the ability of LPS or
cytokines to induce NF-B activation has been shown to be
dependent on the generation of cellular redox stress, decreased GSH
levels could lead to enhanced response of NF-B activation, thereby
increasing iNOS induction. Although activation signals initiating
protein kinase signaling cascades that result in the
phosphorylation of IB on degradation by the
proteosome could be boosted by depleted GSH levels with HNE treatment,
HNE does selectively block signaling events that are required for
IB- degradation. Therefore, it is unlikely that HNE permits
NF-B to translocate to the nucleus and bind its recognition elements
within the promoter regions of NF-B responsive genes, including
iNOS.
It has been reported that HNE was found to be toxic to cells at high concentrations, whereas lower concentrations induced cell growth and DNA synthesis.35 36 37 Similar effects have been observed with other oxidation products, such as H2O2 and oxidized LDL, which, over a narrow concentration range, can cause proliferative and cytotoxic effects.38 39 HNE may exhibit concentration-dependent effects, modulating the expression of different genes at different concentrations. This may be relevant in atherogenesis, in which cell growth, apoptosis, and necrosis are thought to contribute equally to lesion formation.
In conclusion, our data suggest that the aldehyde HNE is one
of the active components in oxidized LDL responsible for the
inhibitory capacities of this lipoprotein on the NF-B
system. Treatment with HNE appears to selectively block signaling
events that are required for IB- degradation, thereby preventing
NF-B activation. Inhibition of NF-B–regulated gene expression,
especially modulation of NO production, may contribute to
certain stages of atherogenesis.
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Acknowledgments |
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This work was supported in part by a grant from Japan Private School Promotion Foundation and by grants-in-aid from the Ministry of Education, Science, and Culture.
Received February 28, 2001; accepted March 6, 2001.
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References |
|---|
|
TopAbstractIntroductionMethodsResultsDiscussionReferences |
|---|
1. Yla-Herttuala S, Palinski W, Rosenfeld ME, Parthasarathy S, Carew TE, Butler S, Witztum JL, Steinberg D. Evidence for the presence of oxidatively modified low density lipoprotein in atherosclerotic lesions of rabbit and man. J Clin Invest. 1989;84:1086–1095.
2.
Palinski W,
Rosenfeld ME, Yla-Herttuala S, Gurtner GC, Socher SS, Butler SW,
Parthasarathy S, Carew TE, Steinberg D, Witztum JL. Low density
lipoprotein undergoes oxidative modification in vivo.
Proc Natl Acad Sci
U S A. 1989;86:1372–1376.
3. Esterbauer H, Jurgens G, Quehenberger O, Koller E. Autoxidation of human low density lipoprotein: loss of polyunsaturated fatty acids and vitamin E and generation of aldehydes. J Lipid Res. 1987;28:495–509.[Abstract]
4. Esterbauer H, Schaur RJ, Zollner H. Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes. Free Radic Biol Med. 1991;11:81–128.[Medline] [Order article via Infotrieve]
5.
Berliner JA, Navab
M, Fogelman AM, Frank JS, Demer LL, Edwards PA, Watson AD, Lusis AJ.
Atherosclerosis: basic mechanisms: oxidation,
inflammation, and genetics.
Circulation. 1995;91:2488–2496.
6. Brand K, Page S, Rogler G, Bartsch A, Brandl R, Knuechel R, Page M, Kaltschmidt C, Baeuerle PA, Neumeier D. Activated transcription factor nuclear factor-kappa B is present in the atherosclerotic lesion. J Clin Invest. 1996;97:1715–1722.[Medline] [Order article via Infotrieve]
7. Baeuerle PA, Baltimore D. NF-kappa B: ten years after. Cell. 1996;87:13–20.[Medline] [Order article via Infotrieve]
8. Sherman MP, Aeberhard EE, Wong VZ, Griscavage JM, Ignarro LJ. Pyrrolidine dithiocarbamate inhibits induction of nitric oxide synthase activity in rat alveolar macrophages. Biochem Biophys Res Commun. 1993;191:1301–1308.[Medline] [Order article via Infotrieve]
9. Mulsch A, Schray-Utz B, Mordvintcev PI, Hauschildt S, Busse R. Diethyldithiocarbamate inhibits induction of macrophage NO synthase. FEBS Lett.. 1993;321:215–218.[Medline] [Order article via Infotrieve]
10.
Xie Q,
Kashiwabara Y, Nathan C. Role of transcription factor NF-kB/Rel in
induction of nitric oxide synthase. J
Biol Chem. 1994;269:4705–4708.
11.
Griscavage JM,
Wilk S, Ignarro LJ. Inhibitors of the proteasome
pathway interfere with induction of nitric oxide synthase in
macrophages by blocking activation of transcription factor
NF-kappa B. Proc Natl Acad Sci
U S A. 1996;93:3308–3312.
12. Nathan C. Nitric oxide as a secretory product of mammalian cells. FASEB J. 1992;6:3051–3064.[Abstract]
13. Lowenstein CJ, Snyder SH. Nitric oxide, a novel biologic messenger. Cell. 1992;70:705–707.[Medline] [Order article via Infotrieve]
14. Moncada S, Higgs EA. Molecular mechanisms and therapeutic strategies related to nitric oxide. FASEB J. 1995;9:1319–1330.[Abstract]
15. Napoli C, D’Armiento FP, Mancini FP, Postiglione A, Witztum JL, Palumbo G, Palinski W. Fatty streak formation occurs in human fetal aortas and is greatly enhanced by maternal hypercholesterolemia: intimal accumulation of low density lipoprotein and its oxidation precede monocyte recruitment into early atherosclerotic lesions. J Clin Invest. 1997;100:2680–2690.[Medline] [Order article via Infotrieve]
16. Muller K, Hardwick SJ, Marchant CE, Law NS, Waeg G, Esterbauer H, Carpentar KL, Mitchinson MJ. Cytotoxic and chemotactic potencies of several aldehydic components of oxidised low density lipoprotein for human monocyte-macrophages. FEBS Lett. 1996;388:165–168.[Medline] [Order article via Infotrieve]
17. Leonarduzzi G, Scavazza A, Biasi F, Chiarpotto E, Camandola S, Vogel S, Dargel R, Poli G. The lipid peroxidation end product 4-hydroxy-2,3-nonenal up-regulates transforming growth factor beta1 expression in the macrophage lineage: a link between oxidative injury and fibrosclerosis. FASEB J. 1997;11:851–857.[Abstract]
18. Camandola S, Scavazza A, Leonarduzzi G, Biasi F, Chiarpotto E, Azzi A, Poli G. Biogenic 4-hydroxy-2-nonenal activates transcription factor AP-1 but not NF-kappa B in cells of the macrophage lineage. Biofactors. 1997;6:173–179.[Medline] [Order article via Infotrieve]
19. Blanc EM, Kelly JF, Mark RJ, Waeg G, Mattson MP. 4-Hydroxynonenal, an aldehydic product of lipid peroxidation, impairs signal transduction associated with muscarinic acetylcholine and metabotropic glutamate receptors: possible action on G alpha(q/11). J Neurochem. 1997;69:570–580.[Medline] [Order article via Infotrieve]
20.
Page S, Fischer
C, Baumgartner B, Haas M, Kreusel U, Loidl G, Hayn M, Ziegler-Heitbrock
HW, Neumeier D, Brand K. 4-Hydroxynonenal prevents NF-kappaB activation
and tumor necrosis factor expression by inhibiting IkappaB
phosphorylation and subsequent proteolysis.
J Biol Chem. 1999;274:11611–11618.
21.
Chamley-Campbell
J, Campbell GR, Ross R. The smooth muscle cell in culture.
Physiol Rev. 1979;59:1–61.
22. Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987;162:156–159.[Medline] [Order article via Infotrieve]
23.
Gross SS, Levi R.
Tetrahydrobiopterin synthesis: an absolute requirement for
cytokine induced nitric oxide generation by vascular smooth
muscle. J Biol Chem. 1992;267:25722–25729.
24. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983;65:55–63.[Medline] [Order article via Infotrieve]
24. Nunokawa Y, Ishida N, Tanaka S. Cloning of inducible nitric oxide synthase in rat vascular smooth muscle cells. Biochem Biophys Res Commun. 1993;191:89–94.[Medline] [Order article via Infotrieve]
25.
Hattori Y,
Hattori S, Kasai K. Troglitazone upregulates nitric oxide synthesis in
vascular smooth muscle cells.
Hypertension. 1999;33:943–948.
26. Smith J, Gross SS. Cloning and regulation of the rat GTP cyclohydrolase gene: coordinate transcriptional induction with iNOS by immuno-stimulants. In: Moncada S, Stamler J, Gross S, Higgs EA, eds. The Biology of Nitric Oxide, Part 5. London, UK: Portland Press; 1996:8.
27.
Hattori Y, Banba
N, Gross SS, Kasai K. Glycated serum albumin-induced nitric
oxide production in vascular smooth muscle cells by nuclear
factor B-dependent transcriptional activation of inducible nitric
oxide synthase. Biochem Biophys Res
Commun. 1999;259:128–132.[Medline]
[Order article via Infotrieve]
28. Thanos D, Maniatis T. NF-kappa B: a lesson in family values. Cell. 1995;80:529–532.[Medline] [Order article via Infotrieve]
29.
Verbeuren TJ,
Jordaens FH, Zonnekeyn LL, Van Hove CE, Coene MC, Herman AG. Effect of
hypercholesterolemia on vascular reactivity in
the rabbit, I: endothelium-dependent and
endothelium-independent contractions and relaxations in
isolated arteries of control and hypercholesterolemic
rabbits. Circ Res. 1986;58:552–564.
30. Ludmer PL, Selwyn AP, Shook TL, Wayne RR, Mudge GH, Alexander RW, Ganz P. Paradoxical vasoconstriction induced by acetylcholine in atherosclerotic coronary arteries. N Engl J Med. 1986;315:1046–1051.[Abstract]
31. Martinez MC, Bosch-Morell F, Raya A, Roma J, Aldasoro M, Vila J, Lluch S, Romero FJ. 4-Hydroxynonenal, a lipid peroxidation product, induces relaxation of human cerebral arteries. J Cereb Blood Flow Metab. 1994;14:693–696.[Medline] [Order article via Infotrieve]
32. Romero FJ, Romero MJ, Bosch-Morell F, Martinez MC, Medina P, Lluch S. 4-Hydroxynonenal-induced relaxation of human mesenteric arteries. Free Radic Biol Med. 1997;23:521–523.[Medline] [Order article via Infotrieve]
33.
Uchida K,
Shiraishi M, Naito Y, Torii Y, Nakamura Y, Osawa T. Activation of
stress signaling pathways by the end product of lipid peroxidation:
4-hydroxy-2-nonenal is a potential inducer of intracellular peroxide
production. J Biol
Chem. 1999;274:2234–2242.
34.
Cosentino F,
Katusic ZS. Tetrahydrobiopterin and dysfunction of
endothelial nitric oxide synthase in coronary
arteries. Circulation. 1995;91:139–144.
35.
Krokan H,
Grafstrom RC, Sundqvist K, Esterbauer H, Harris CC. Cytotoxicity, thiol
depletion and inhibition of O6-methylguanine-DNA methyltransferase by
various aldehydes in cultured human bronchial fibroblasts.
Carcinogenesis. 1985;6:1755–1759.
36. Tessitore L, Matera L, Bonelli G, Baccino FM, Dianzani MU. Aliphatic aldehydes inhibit the proliferative response of human peripheral blood lymphocytes to phytohemagglutinin and alloantigens. Chem Biol Interact. 1987;62:217–226.[Medline] [Order article via Infotrieve]
37.
Ruef J, Rao GN,
Li F, Bode C, Patterson C, Bhatnagar A, Runge MS. Induction of rat
aortic smooth muscle cell growth by the lipid peroxidation product
4-hydroxy-2-nonenal.
Circulation. 1998;97:1071–1078.
38. Chatterjee S. Role of oxidized human plasma low density lipoproteins in atherosclerosis: effects on smooth muscle cell proliferation. Mol Cell Biochem. 1992;111:143–147.[Medline] [Order article via Infotrieve]
39.
Guyton KZ, Liu Y,
Gorospe M, Xu Q, Holbrook NJ. Activation of mitogen-activated
protein kinase by H2O2:
role in cell survival following oxidant injury.
J Biol Chem. 1996;271:4138–4142.
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