From the Graduate Institute of Life Sciences and Departments of Medical
Research (T.-C.C.), Pharmacology (M.-H.Y.), Anesthesiology (C.-Y.L.),
Tri-Service General Hospital; the National Defense Medical Center; and the
Division of Cardiology, Veterans General Hospital (Y.-A.D.), Taipei, Republic
of China.
Correspondence to Yu-An Ding, Division of Cardiology, Veterans General Hospital, 201, Sec. 2, Shih-Pai Rd. Taipei, Taiwan, ROC.
Considerable evidence demonstrates that NO plays a critical role in the
maintenance of blood pressure
homeostasis.7 8 In addition, NO also prevents the
activation and adhesion of platelets and neutrophils to the
endothelium. NO induces vascular smooth muscle
relaxation by activation of soluble guanylate cyclase,
which leads to an increase of cGMP, causing a decrease in intracellular
Ca2+ with subsequent
vasodilation.9 Therefore, alterations in NO
synthesis may be related to the pathogenesis of hypertension. This
hypothesis was supported by the finding that the vasodilatory response
to ACh was impaired in SHR when compared with
WKY,10 11 12 presumably due to a decrease in
synthesis and/or release of endothelium-derived
relaxation factor. However, the effects of aging and hypertension on
the activity and protein expression of NOS have not been investigated.
Here we report direct measurement of the changes of NOS activity and
protein expression of aortas at different age groups of the SHR and its
control group, WKY. To further clarify the relationship between
hypertension and the changes of NOS expression in SHR, we treated SHR
with an antihypertensive agent, the ACEI quinapril, evaluating its
effect on NOS expression.
NOS Activity Assay
Western Blotting
Measurement of Plasma Nitrite/Nitrate Concentration
Measurement of Plasma Levels of TNF-
In Vivo Study of Quinapril
Statistical Analysis
Activity of eNOS and iNOS in the Aorta
Western Analysis of eNOS and iNOS in the Aorta
Plasma NO2-/NO3-
and TNF-
Similarly, the basal plasma TNF-
Effects of Quinapril on Activity and Expression of iNOS
In the 14-to-17- and 63-week groups, the basal eNOS activity of SHR was
significantly lower than that of WKY, which was paralleled with a
concurrent fall in eNOS protein expression. The changes of eNOS were
associated with an elevation of blood pressure. There is evidence that
there are no differences in eNOS activity and protein expression
between SHR and WKY in the prehypertensive state. Many studies have
reported that hypertension is associated with decreased
endothelium-dependent
relaxations.12 15 This has been demonstrated not
only in large conduit arteries but also in resistant arteries
obtained from hypertensive rats.16 17 The cause
of the reduction of eNOS activity and expression in SHR is still
unclear. It has been proposed that the damage of
endothelium structure and/or functions may be an
important factor, and this defect is closely related to hypertension.
This hypothesis is supported by these findings, indicating that
endothelial function is still normal in SHR at 4 weeks
old but abnormal at 14 weeks of age when hypertension is
established.18 19 Furthermore, because
antihypertensive therapy with calcium channel antagonists
or ACEI can improve the endothelium-dependent
relaxation to ACh,20 it is likely that the
endothelial dysfunction is a consequence of high blood
pressure. However, the possibility that the endothelial
dysfunction may also be a cause of hypertension cannot be ruled
out.
Apart from hypertension, aging may be another important factor causing
abnormal expression of eNOS. Evidence was that our results demonstrated
that aged WKY possessed lower eNOS activity than that of adult WKY, but
no significant difference was found in the same age-matched SHR. These
findings were consistent with the results of Küng and
Lüscher,21 showing that senescent WKY
(72-week-old) exhibited a reduced endothelium-dependent
relaxation to ACh in aortas compared with that of 12-week-old WKY, but
this change was not observed in aged SHR. The reasons are unclear.
However, it is possible that the structure and/or function of the
endothelium in the adult SHR has been impaired and that
it is not being further damaged at a more aged state.
Our previous study showed that in endothelium-denuded
preparations, L-arginine has no effect on rings from
16-week-old WKY rats but causes a slight relaxation in those from SHR.
After treatment of rats with LPS, the L-arginine–induced
relaxation was observed in both strains, with SHR showing a greater
response.22 In addition, the
L-arginine–induced relaxation was inhibited by
aminoguanidine, a relatively selective inhibitor of iNOS,
in both strains (after LPS treatment) or in SHR only (before LPS
treatment). It is well known that if iNOS is induced, the NO
production from VSMC is mainly dependent on the entry of
extracellular L-arginine. Therefore, it is reasonable to
expect that L-arginine would evoke relaxations in both
strains treated with LPS. These findings suggest that iNOS is
present in the SHR, which is further enhanced by the stimulation
with LPS, as evidenced by the results of activity and Western blot
analysis of iNOS in SHR under basal conditions and after LPS
challenge. In addition, Lee and Webb23
demonstrated that the addition of L-arginine caused a
greater relative reversal of L-NMMA–induced inhibition of ACh-induced
relaxation in stroke-prone SHR, suggesting that this greater relaxation
is mediated by iNOS, consistent with our findings. Furthermore,
the VSMC of aorta and macrophages from SHR can produce greater
amounts of NO stimulated by LPS than that from
WKY,24 but VSMC of mesenteric artery induced by
interleukin-1ß and endothelial cells of
aorta15 released less NO compared with those from
WKY, showing the heterogeneity of vascular
beds.25 Another finding was that the basal
activity and expression of iNOS appeared in aged WKY, which may result
from aging or vascular injury. The causes for the increased activity
and expression of iNOS in SHR are still unknown.
Although the pathophysiological significance of
iNOS in regulation of blood pressure is unclear, we propose that the
appearance of iNOS in SHR may reflect the pathological state of
vessels, presumably a consequence of injury due to hypertension. This
is strongly supported by the finding that the exaggerated iNOS
expression can be attenuated after reduction of blood pressure with
chronic treatment of quinapril (Fig 4
It is well known that cytokines such as TNF-
In conclusion, an early decline (from 4 to 14 weeks) in eNOS activity
and protein expression was found in SHR but not in WKY. In addition, in
the aging process (from 14 to 63 weeks), the decreased eNOS activity
was observed in WKY, but no further decrease was observed in SHR. In
contrast, the changes of iNOS activity and protein expression were
opposite to those of eNOS in WKY and SHR at 14 and 63 weeks, suggesting
that the alteration of eNOS and iNOS is associated with aging and the
development of hypertension. From the development of the hypertensive
state (14 weeks), the iNOS in the aorta is present in SHR and is
further enhanced by stimulation with LPS. The higher basal levels of
NO2-/NO3-
may result from higher expression of iNOS in SHR. We proposed that the
pathological condition of vessels in hypertension may be responsible
for an increase of iNOS expression. In addition, the abnormal
expression of iNOS was attenuated by reduction of blood pressure with
quinapril, which further indicates that hypertension may be an
important factor in the regulation of iNOS expression. However, the
"cause and effect" relationship between hypertension and iNOS
expression needs to be further clarified.
Received June 9, 1997;
first decision July 3, 1997;
accepted October 3, 1997.
© 1998 American Heart Association, Inc.
Scientific Contributions
Alterations of Nitric Oxide Synthase Expression With Aging and Hypertension in Rats
Abstract
Top
Abstract
Introduction
Methods
Results
Discussion
References
Abstract—The activity and protein
expression of endothelial nitric oxide synthase (eNOS)
and inducible NOS (iNOS) were investigated during the development of
hypertension in spontaneously hypertensive rats (SHR). SHR and
Wistar-Kyoto rats (WKY) were studied at three different ages: 4, 14 to
17, and 63 weeks of age. After treatment with saline or
lipopolysaccharide (LPS, 10 mg/kg IV) for 3 hours, the aortas
were removed for measurement of NOS activity and protein expression
assay by [3H]-L-citrulline formation method
and Western blot analysis, respectively. Plasma levels of
nitrite/nitrate
(NO2-/NO3-) and tumor
necrosis factor-
(TNF-) were also determined. At 14 to 17 weeks
and 63 weeks, the basal activity and protein expression of eNOS in the
aortas were significantly lower in SHR than in WKY. In addition, the
aged WKY exhibited lower eNOS activity than that of adult WKY, but this
change was not seen in SHR. By comparison, the basal activity and
protein expression of iNOS were only observed in SHR of the
14-to-17-week group and in the 63-week group; SHR still exhibited
higher activities, and these differences were further exaggerated by
treatment with LPS. The basal and LPS-induced
NO2-/NO3- and TNF-
levels in the plasma were also higher in the SHR except the 4-week
group. After treatment with quinapril, the basal and LPS-induced
expressions of iNOS in SHR were significantly attenuated. Our results
demonstrated that alterations of activity and protein expression of
eNOS and iNOS occurred in SHR. In addition, aging may reduce the
activity of eNOS in WKY but not in SHR. The decline of eNOS activity
and/or expression may contribute to the development of hypertension,
whereas the increase of iNOS expression may be a consequence of the
pathological state of vessels associated with hypertension in SHR.
However, the augmented expression of iNOS in SHR was attenuated by
antihypertensive therapy, suggesting that the abnormal expression of
iNOS is associated with hypertension.
Key Words: nitric oxide synthase • hypertension, experimental • rats, inbred SHR
Introduction
Top
Abstract
Introduction
Methods
Results
Discussion
References
The generation of NO
from L-arginine by NOS is an important autocrine and
paracrine signaling pathway in the regulation of various cell functions
and in cell-cell communication.1 2 NO, also known
as endothelium-derived relaxation factor, is produced
endogenously in various types of
cells.2 Molecular and enzymatic studies have
indicated that there are at least two major isoforms of NOS: eNOS and
iNOS, found in macrophages, VSMC, and other
tissues,3 and the expression can be induced by
bacterial endotoxin (LPS) and cytokines such as
interleukin-1ß, TNF-,4 or by mechanical,
viral, or bacterial injuries.5 NO produced from
the iNOS acts as a cytotoxic agent against tumor cells, bacteria,
fungi, and protozoa and plays an important role in pathological
vasodilation and host tissue damage in endotoxic shock or altered
immunologic status.6
Methods
Top
Abstract
Introduction
Methods
Results
Discussion
References
In this study 81 male SHR and 60 male WKY were divided into
three age groups: 4, 14 to 17, and 63 weeks old. Systolic blood
pressure was measured in warmed, restrained, unanesthetized
animals using the caudal artery tail-cuff MOD 59 plethysmograph (blood
pressure meter/amplifier, Itic Inc). Animals were anesthetized
with urethane (0.6 g/kg IP), and the carotid artery was cannulated for
blood collection. A jugular vein was also cannulated for
intravenous bolus administration of vehicle (saline) or LPS
(10 mg/kg). After treatment for 3 hours, the aorta was removed and
stored at -70°C for the determination of NOS activity and
protein expression.
Aortas were homogenized on ice with a polytron
(model PT MR3000, Littau) according to the method of Klemm et
al.13 NOS activity was determined by measuring
the conversion of [3H]–L-arginine
to [3H]–L-citrulline. Tissue
homogenates (approximately 60 µg protein) were incubated
in 20 mmol/L HEPES buffer (pH 7.5) containing 10 µmol/L
L-arginine and
[3H]–L-arginine (3 µCi/mL),
L-valine (60 mmol/L), NADPH (1 mmol/L),
calmodulin (30 nmol/L), tetrahydrobiopterin (5
µmol/L), and calcium (2 mmol/L) for 20 minutes at 37°C.
Reaction was stopped by adding 1 mL of ice-cold HEPES buffer (pH 5.5)
containing EGTA (2 mmol/L) and EDTA (2 mmol/L) and then
applying to Dowex 50W (Na+ form) columns, and the
amount of [3H]–L-citrulline eluted
was quantified by liquid scintillation counter (Beckman, model LS3801).
The activity of the Ca2+-dependent NOS was
determined from the difference between the
[3H]–L-citrulline produced from
samples containing 2 mmol/L calcium and samples without calcium
and with EGTA (2 mmol/L); the activity of the iNOS was determined
from the difference between samples containing 2 mmol/L EGTA and
samples without NADPH.13 Protein determination
was made using dye-binding assay (Bio-Rad) with bovine serum
albumin as a standard.
Aorta lysate containing 10 µg protein was denatured and
an equal amount of protein loaded on 7.5% SDS-polyacrylamide
gel and then was transferred to nitrocellulose membranes using
PharmSystem (Pharmacia Biotech). The membrane was blocked with 1%
bovine serum albumin in Tris-buffer solution (TBS; pH 8.0)
containing 0.1% Tween-20 for 2 hours at room temperature. Next, the
membrane was incubated overnight at 4°C with mouse monoclonal
anti-iNOS or anti-eNOS antibody (1:2000 dilution, Transduction
Laboratories) in TBS containing 0.1% Tween-20. The membrane was washed
and finally incubated with a 1:1000 dilution of anti-mouse IgG
conjugated to horseradish peroxidase for 1 hour at room temperature.
After successive washes with TBS, the immunocomplexes were detected
using an enhanced horseradish peroxidase/luminol chemiluminescence
reaction (Amersham International plc) and exposed to x-ray film for 3
to 5 minutes. The density of respective bands was quantified by
densitometric scanning of the Western blots using Image-Pro Plus
software. We took the density of the band representing 1
µg eNOS or iNOS positive control purified from human
endothelial cells and mouse macrophages lysate
(Transduction Laboratories), respectively, as 100% to calculate the
relative density of other bands on the same gel.
Plasma (100 µL) was mixed with 160 µL of 75 mmol/L
ZnSO4 and 140 µL of 80 mmol/L NaOH for 10
minutes and centrifuged at 6000g for 10 minutes to
deproteinize the plasma. The supernatant (100 µL) was removed and
incubated with 25 µL substrate buffer (NADPH, 0.6 mmol/L;
flavine adenine dinucleotide, 5 µmol/L; nitrate
reductase, and 2 U/mL pH 7.6 at 37°C for 1 hour to convert
NO3- to
NO2-. The total nitrite
(NO2-/NO3-)
assay was determined by the method of Misko et
al.14 The intensity of fluorescence was
measured using a luminescence spectrometer (model LS-5, Perkin-Elmer)
with excitation at 365 nm and emission at 450 nm. Nitrite levels in
samples were calculated using a standard curve of nitrite to which
NADPH had been added.
Rats were anesthetized and instrumented as described
above. Blood samples (0.5 mL) were taken at 0 and 90 minutes after the
injection of LPS. The amount of TNF- in the plasma was measured by
using a rat TNF- ELISA kit (Genzyme Corporation).
SHR (14 to 17 weeks) were divided into two groups: one was
treated with quinapril (10 mg/kg/day, PO) for 2 weeks, the other was
treated with saline to serve as control. Each group was further divided
into two groups, treated with or without LPS challenge. After the
administration of LPS (10 mg/kg, IV) for 3 hours, the activity and
protein expression of iNOS in aorta homogenates and plasma
concentrations of
NO2-/NO3-
and TNF- (at 90 minutes) were measured according to above
methods.
All data were analyzed by one-way ANOVA, and statistical
significance was assessed by Student's unpaired two-tailed
t test. Differences were considered statistically
significant when the value was P<.05.
Results
Top
Abstract
Introduction
Methods
Results
Discussion
References
Blood Pressure and Body Weight
The systolic blood pressure (mm Hg) of SHR was
significantly higher than that of age-matched WKY except at 4 weeks of
age (prehypertensive state). The body weight was similar between SHR
and WKY in each age-matched group (Table 1
).
View this table:
[in a new window]
Table 1. Characteristics of Study Rats at 4, 14 to 17 and 63
Weeks of Age
In the prehypertensive state, the basal activity of eNOS
(Ca2+-dependent) in the aorta was similar in SHR
and WKY. However, in the 14-to-17- and 63-week groups, the activity of
eNOS in SHR was greatly attenuated when compared with age-matched WKY.
In addition, in WKY, the eNOS activity of the 63-week group was
significantly lower than that in the 14-to-17-week group. In
comparison, at young age (4 weeks), iNOS activity
(Ca2+-independent) of aorta was undetectable in
both SHR and WKY. It is important to note that in the 14-to-17-week
group, the basal activity of iNOS was only observed in SHR. However, at
elder age (63 weeks), the activity of iNOS also appeared in WKY, but it
was much lower than that in SHR. On stimulation with LPS, the activity
of iNOS of both strains was further enhanced, and the activity of iNOS
in SHR remained higher than that in WKY except for the 4-week group
(Table 2).
View this table:
[in a new window]
Table 2. NOS Activity in Aorta From WKY and SHR
The basal protein expression of eNOS in the aorta was similar in
the 4-week group of both strains. However, it was significantly lower
in SHR than in WKY at 14 to 17 and 63 weeks of age (Fig 1). Although in WKY the eNOS protein
expression in the 63-week group was slightly lower than that in the
14-to-17-week group, the difference did not reach statistical
significance. Interestingly, the basal protein expression of iNOS was
only observed in SHR of the 14-to-17-week group; it was not detectable
in the prehypertensive state in SHR and WKY. At 63 weeks, the protein
expression of iNOS in WKY increased slightly but remained significantly
lower than that in SHR. After LPS stimulation, the protein expression
of iNOS was further elevated and remained significantly higher in SHR
than in WKY in the 14-to-17- and 63-week groups (Fig 2). In addition, these observations were
paralleled with the findings of NOS activity studies.
View larger version (31K):
[in a new window]
Figure 1. Western blot analysis of basal eNOS in
aorta homogenates in SHR and WKY of different age groups.
Analysis of eNOS protein expression in aorta
homogenates was performed with a mouse monoclonal anti-eNOS
antibody. An equal amount of protein was loaded in each column. The
respective density of bands was quantified by densitometric scanning,
and the relative density was calculated by taking the density of 1 µg
eNOS positive control band as 100%. Data were mean±SEM of 5 to 6 rats
in each group. **P<.01 compared with age-matched
WKY.
View larger version (28K):
[in a new window]
Figure 2. Western blot analysis of iNOS in aorta
homogenates under basal or LPS-induced condition in WKY and
SHR of different age groups. Analysis of iNOS protein
expression in aorta homogenates from WKY and SHR (4,
14–17, and 63 weeks old) under basal conditions (A) or after treated
with LPS (10 mg/kg IV, for 3 hours; B) was performed with a monoclonal
anti-iNOS antibody generated against murine macrophage iNOS. An
equal amount of protein was loaded in each column. The respective
density of bands was quantified by densitometric scanning, and the
relative density was calculated by taking the density of 1 µg iNOS
positive control band as 100%. Data are mean±SEM of 5 to 6 rats in
each group. *P<.05, **P<.01 compared
with age-matched WKY. ##P<.01 compared with the
respective value in the 14-to-17-week group. Levels
The basal plasma
NO2-/NO3-
level was significantly greater in SHR than in WKY in the 14-to-17- and
63-week groups. After administration with LPS for 3 hours, the plasma
NO2-/NO3-
level was markedly increased in both strains; however, the elevation of
NO2-/NO3-
was higher in SHR than in WKY in the 14-to-17- and 63-week groups
(Table 3).
View this table:
[in a new window]
Table 3. Plasma
NO2-/NO3- Levels in
WKY and SHR Under Basal or LPS-Treated Condition level was higher in SHR than in WKY
in the 14-to-17- and 63-week groups (Fig 3A). The injection of LPS caused a
bell-shape change in plasma TNF- level that reached a peak value at
90 minutes; thereafter, it returned to the pretreatment level at 240
minutes.13 Therefore, the 90-minute time point
was chosen for our study. The increase of TNF- induced by LPS was
also much greater in SHR than in WKY at 14-to-17- and 63-week groups
(Fig 3B).
View larger version (17K):
[in a new window]
Figure 3. Comparison of the levels of plasma TNF- under
basal conditions or after LPS treatment between WKY and SHR at
different times (age groups). A, Basal levels of plasma TNF- in WKY
and SHR (n=9 to 10 for each group). B, Plasma TNF- levels at 90
minutes after LPS (10 mg/kg IV) injection in WKY and SHR (n=8 to 11 for
each group). *P<.05, **P<.01 compared
with the age-matched WKY.
After treatment with quinapril for 2 weeks, the systolic
blood pressure of SHR was significantly reduced from 195±2 to
161±3 mm Hg (P<.01, n=7). Concurrently, the basal
and LPS-induced activity and protein expression of iNOS were also
significantly attenuated by quinapril treatment (Fig 4) and accompanied by a significant
decrease in basal plasma
NO2-/NO3-
(4.83±0.18 to 3.75±0.12 µmol/L, P<.05, n=7) and
TNF- (253±21 to 190±17 pg/mL, P<.05, n=7) levels when
compared with the values of untreated group.
View larger version (17K):
[in a new window]
Figure 4. Changes of activity and protein expression of iNOS
in the aortas from 14-to-17-week-old SHR after treatment with quinapril
for 2 weeks. A, Changes of basal or LPS-treated (10 mg/kg IV, for 3
hours) activity of iNOS in aorta homogenates obtained from
SHR treated with normal saline (control) or quinapril (10 mg/kg PO) for
2 weeks. B, Changes of basal or LPS-treated (10 mg/kg IV, for 3 hours)
protein expression of iNOS in aorta homogenates obtained
from SHR treated with normal saline (control, 
) or quinapril (10
mg/kg PO, 
) for 2 weeks. An equal amount of protein was loaded in
each column. The density of respective bands was quantified by
densitometric scanning, and the relative density was calculated taking
the density of 1 µg iNOS positive control band as 100%. Data are
mean ±SEM of 5 to 6 rats in each group. **P<.01,
*P<.05 compared with respective control group.
Discussion
Top
Abstract
Introduction
Methods
Results
Discussion
References
In the development of hypertension, the SHR exhibits a decrease in
the activity and protein expression of eNOS in the aorta with an
elevation of activity and protein expression of iNOS, accompanied by
greater plasma
NO2-/NO3-
and TNF- levels when compared with those from age-matched WKY. In
addition, aged WKY exhibited a significantly lower eNOS activity than
that in adult WKY. The augmented expression of iNOS in SHR was
attenuated when blood pressure was reduced with quinapril. 
). In addition, antihypertensive
therapy with ACEI or a calcium channel antagonist can also
improve the endothelial-dependent relaxation to
ACh.26 27 These results suggest that hypertension
is an important factor in the modulation of eNOS and/or iNOS expression
in vascular beds. The mechanisms by which quinapril reduces iNOS
expression are still unclear. Recently, some studies have demonstrated
that angiotensin II stimulates superoxide formation by
activating NADPH and NADH oxidase both in cultured
VSMC28 and in in vivo
studies.29 Reactive oxygen species have been
implicated in the pathogenesis of hypertension and tissue
damage.30 Enhanced oxygen species has been
reported in the microvascular wall and endothelial
cells of SHR.31 32 Furthermore, oxidative stress
can induce transcription factor nuclear factor-
B–responsive genes,
such as iNOS.33 Therefore, it is possible that
the elevated iNOS observed in SHR may have been induced by the
increased oxidative stress. These observations suggest that the
mechanisms by which ACEI attenuates the iNOS expression and corrects
the polyploidy of VSMC in SHR may be also associated with the reduction
of angiotensin II–induced superoxide production.
As we have mentioned above, in SHR with lower eNOS and higher iNOS, the
sum of NO production may be unchanged or even enhanced.
Therefore, the higher basal and LPS-induced
NO2-/NO3-
levels in SHR were most likely mediated by iNOS rather than by eNOS.
Despite the report by Sawada et al34 that release
of NO in response to ACh is unaltered in SHR, the possibility that NOS
in blood vessels is changed in SHR cannot be ruled out. and
interleukin-1ß secreted by macrophages or VSMC are potent
stimulators for iNOS. Our results showed that the plasma TNF- level
was higher in SHR both under basal conditions or LPS treatment,
suggesting that an altered immune response may exist in
SHR.35 In addition, according to our previous
hemodynamic results,22 the
overproduction of TNF- may play an important role in the
more severe hypotensive effect induced by LPS and in activation of iNOS
in SHR.
Selected Abbreviations and Acronyms
ACEI
=
angiotensin-converting enzyme inhibitor
ACh
=
acetylcholine
eNOS
=
Ca2+/calmodulin-dependent NOS expressed in
endothelial cells
iNOS
=
inducible Ca2+/calmodulin-independent NOS
LPS
=
lipopolysaccharide
NOS
=
nitric oxide synthase
SBP
=
systolic blood pressure
SHR
=
spontaneously hypertensive rats
TNF-
=
tumor necrosis factor-
VSMC
=
vascular smooth muscle cells
WKY
=
Wistar-Kyoto rats
Acknowledgments
This work was supported by a grant from the National Science
Council of Republic of China (NSC86–2314-B016–048). We thank
Shu-Chuan Lin and Kuei-Ying Ho for their skillful technical assistance.
The authors also thank Dr Peter Chiu (Schering-Plow Research Institute,
Kenilworth, UK) and Dr Chin-Chen Wu (Department of Pharmacology) for
their helpful comments.
References
Top
Abstract
Introduction
Methods
Results
Discussion
References
B DNA binding and inducible NOS mRNA in human
epithelial cells. Biochem Biophys Res Comm. 1994;199:1518–1524.[Medline]
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K. Yamamoto, K. Takeshita, T. Kojima, J. Takamatsu, and H. Saito Aging and plasminogen activator inhibitor-1 (PAI-1) regulation: implication in the pathogenesis of thrombotic disorders in the elderly Cardiovasc Res, May 1, 2005; 66(2): 276 - 285. [Abstract] [Full Text] [PDF]
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 L. L. Kolo, T. C. Westfall, and H. Macarthur Modulation of neurotransmitter release by NO is altered in mesenteric arterial bed of spontaneously hypertensive rats Am J Physiol Heart Circ Physiol, October 1, 2004; 287(4): H1842 - H1847. [Abstract] [Full Text] [PDF]
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 G. Spinetti, M. Wang, R. Monticone, J. Zhang, D. Zhao, and E. G. Lakatta Rat Aortic MCP-1 and Its Receptor CCR2 Increase With Age and Alter Vascular Smooth Muscle Cell Function Arterioscler. Thromb. Vasc. Biol., August 1, 2004; 24(8): 1397 - 1402. [Abstract] [Full Text] [PDF]
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R. Hernanz, A. M. Briones, M. J. Alonso, E. Vila, and M. Salaices Hypertension alters role of iNOS, COX-2, and oxidative stress in bradykinin relaxation impairment after LPS in rat cerebral arteries Am J Physiol Heart Circ Physiol, July 1, 2004; 287(1): H225 - H234. [Abstract] [Full Text] [PDF]
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M. G Ferrini, H. H Davila, E. G.A Valente, N. F Gonzalez-Cadavid, and J. Rajfer Aging-related induction of inducible nitric oxide synthase is vasculo-protective to the arterial media Cardiovasc Res, March 1, 2004; 61(4): 796 - 805. [Abstract] [Full Text] [PDF]
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S. Biro, A. Masuda, T. Kihara, and C. Tei Clinical Implications of Thermal Therapy in Lifestyle-Related Diseases Experimental Biology and Medicine, November 1, 2003; 228(10): 1245 - 1249. [Abstract] [Full Text] [PDF]
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 D. E. Berkowitz, R. White, D. Li, K. M. Minhas, A. Cernetich, S. Kim, S. Burke, A. A. Shoukas, D. Nyhan, H. C. Champion, et al. Arginase Reciprocally Regulates Nitric Oxide Synthase Activity and Contributes to Endothelial Dysfunction in Aging Blood Vessels Circulation, October 21, 2003; 108(16): 2000 - 2006. [Abstract] [Full Text] [PDF]
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G. G. Geary and J. N. Buchholz Selected Contribution: Effects of aging on cerebrovascular tone and [Ca2+]i J Appl Physiol, October 1, 2003; 95(4): 1746 - 1754. [Abstract] [Full Text] [PDF]
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M. E. Safar and P. Laurent Pulse pressure and arterial stiffness in rats: comparison with humans Am J Physiol Heart Circ Physiol, October 1, 2003; 285(4): H1363 - H1 |