J Am Soc Nephrol 13: 1788–1794, 2002
Impaired Regulation of Renal Oxygen Consumption in
Spontaneously Hypertensive Rats
STEPHEN ADLER and HARER HUANG
New York Medical College, Department of Medicine, Division of Nephrology, Hawthorne, New York.
Abstract. Abnormalities of nitric oxide (NO) and oxygen rad-
ical synthesis and of oxygen consumption have been described
in the spontaneously hypertensive rat (SHR) and may contrib-
ute to the pathogenesis of hypertension. NO plays a role in the
regulation of renal oxygen consumption in normal kidney, so
the response of renal cortical oxygen consumption to stimula-
tors of NO production before and after the addition of the
superoxide scavenging agent tempol (4-hydroxy-2,2,6,6-tetra-
methyl piperidine-1-oxyl) was studied. Baseline cortical oxy-
gen consumption was similar in SHR and Wistar-Kyoto
(WKY) rats (SHR: 600 ⫾ 55 nmol O2/min per g, WKY: 611
⫾ 51 nmol O2/min per g, P ⬎ 0.05). Addition of bradykinin,
enalaprilat, and amlodipine decreased oxygen consumption
significantly less in SHR than WKY (SHR: bradykinin ⫺13.9
⫾ 1.9%, enalaprilat ⫺15.3 ⫾ 1.6%, amlodipine ⫺11.9 ⫾
Nitric oxide (NO) plays an important role in regulation of
vascular tone. Absence of the NO generating enzyme endothe-
lial nitric oxide synthase (eNOS) or impairment of NO pro-
duction leads to hypertension in animal models and can in-
crease vascular tone in humans (1– 4). In a model of genetic
hypertension in the rat, the spontaneously hypertensive rat
(SHR), abnormalities in synthesis of NO, expression of the
NO-synthesizing enzymes, or both have been described both in
vitro and in vivo, but with conflicting results. Thus, several
studies have been performed to provide evidence of impaired
vasodilation in response to acetylcholine, an effect mediated by
endothelium-derived relaxing factor or NO, decreased eNOS
expression, or decreased NO synthesis in SHR (5–11). These
abnormalities are present as early as 5 wk of age, a period
before the development of hypertension (7). However, several
of these studies have shown a decreased effect of NO rather
than direct evidence of decreased production.
On the other hand, evidence of increased expression of NO
synthesizing enzymes (eNOS and inducible NO synthase),
increased NO production, or both have also been noted (12–
18), both before and after the onset of hypertension. One
Received January 25, 2002. Accepted April 3, 2002.
Correspondence to Dr. Stephen Adler, Division of Nephrology, Department of
Medicine, 19 Bradhurst Avenue, Suite 0100, Hawthorne, NY 10532. Phone:
914 493-7701; Fax: 914 345-0652; E-mail: stephen@nymc.edu
1046-6673/1307-1788
Journal of the American Society of Nephrology
Copyright © 2002 by the American Society of Nephrology
DOI: 10.1097/01.ASN.0000019781.90630.0F
0.7%; WKY: bradykinin ⫺22.8 ⫾ 1.0%, enalaprilat ⫺24.1 ⫾
2.0%, amlodipine ⫺20.7 ⫾ 2.3%; P ⬍ 0.05), consistent with
less NO effect in SHR. Addition of tempol reversed the defects
in responsiveness to enalaprilat and amlodipine, suggesting
that inactivation of NO by superoxide contributes to decreased
NO availability. The response to an NO donor was similar in
both groups and was unaffected by the addition of tempol.
These results demonstrate that NO availability in the kidney is
decreased in SHR, resulting in increased oxygen consumption.
This effect is due to enhanced production of superoxide in
SHR. By lowering intrarenal oxygen levels, reduced NO may
contribute to susceptibility to injury and renal fibrosis. Increas-
ing NO production, decreasing oxidant stress, or both might
prevent these changes by improving renal oxygenation.
possible explanation for this discrepancy is inactivation of NO,
explaining increased production but less effect in SHR. NO is
inactivated by reaction with superoxide (O2⫺) to produce per-
oxynitrite and increased production of superoxide has been
demonstrated in SHR (13,19 –21). Manipulations that reduce
superoxide production have also been shown to lower BP in
these animals (19 –21).
NO also modulates oxygen consumption in the whole animal
and in isolated tissues, such as heart and kidney, with an
inverse relationship between NO production and oxygen con-
sumption (22–24). We have previously shown that NO pro-
duction by the eNOS isoenzyme in the kidney is responsible
for regulation of renal oxygen consumption (25). Whole-body
oxygen consumption is increased in SHR, an observation con-
sistent with a decreased effect of NO (26). More recently,
studies in SHR have demonstrated a relative increase in oxygen
consumption in kidney when compared with sodium reabsorp-
tion, indicating relative inefficiency of oxygen usage in these
animals (27). Therefore, we hypothesized that NO regulation
of oxygen consumption is impaired in the kidney of SHR and
that interference with superoxide production would restore the
normal effects of agonists of NO production on renal oxygen
consumption.
Materials and Methods
Reagents
Bradykinin, enalaprilat, S-nitroso-N-acetylpenicillamine (SNAP),
N-nitro-L-arginine methyl ester (L-NAME), tempol (4-hydroxy-
2,2,6,6-tetramethyl piperidine-1-oxyl), sodium succinate, and sodium
cyanide were purchased from Sigma Chemical Company (St. Louis,
MO). Amlodipine was provided by Pfizer (Groton, CT).
J Am Soc Nephrol 13: 1788–1794, 2002
Animals
Male SHR (n ⫽ 6) and Wistar-Kyoto (WKY; n ⫽ 6) rats were
purchased from Taconic Farms, Inc. (Germantown, NY) when they
were 10 wk old and studied after 1 wk of acclimatization. Rats were
maintained on a standard rat chow with 0.4% sodium content (Lab-
oratory Rodent Diet, Richmond, IN). The rats were allowed free
access to food and water until the day they were killed. Before they
were killed, BP and heart rate were measured with a noninvasive BP
monitor (Columbus Instruments, Columbus, OH), and blood was
drawn from the femoral vein for measurement of hemoglobin, blood
urea nitrogen, and creatinine assessment. After the animals were
killed, the left kidneys were removed, decapsulated, and weighed.
Preparation of Kidney Tissue Slices and Measurement
of O2 Consumption
Thin slices of renal cortex (approximately 1 mm, weight 10 to 20
mg) were prepared and incubated in Krebs bicarbonate solution (con-
taining, in mmol/L, NaCl 118, KCl 4.7, CaCl2 1.5, NaHCO3 25,
KH2PO4 1.2, MgSO4 1.1 and glucose 5.6, pH 7.4) bubbled with 21%
O2/5% CO2/74% N2 at 37°C for 2 h. At the end of incubation, each
piece of tissue was placed in a stirred chamber with 3 ml of air-
saturated Krebs bicarbonate solution containing 10 mmol/L N-2-
hydroxyethylpiperazine-N'-2-ethane sulfonic acid and 5.6 mmol/L
glucose (pH 7.4). The chamber was sealed with a Clark-type platinum
O2 electrode (Yellow Springs Instruments, Yellow Springs, OH). O2
consumption was measured polarographically with an O2 monitor
(model YSI 5300) connected to a linear chart recorder (model 1202,
Barnstead/Thermolyne Corp., Dubuque, IA). Dose-response curves of
the effect of different agonists on kidney O2 consumption were then
measured. Succinate (10⫺3 mol/L) and then sodium cyanide (10⫺3
mol/L) were added at the end of each experiment to confirm that
changes in O2 consumption originated from mitochondrial respiration.
Renal cortical O2 consumption is calculated as the rate of decrease
in O2 concentration, assuming an initial O2 concentration of 224
nmol/ml (calculated from O2 solubility at 37°C and 1 atm pressure)
and is expressed as nanomoles of O2 consumed per minute per gram
of tissue. O2 consumption due to the electrode is less than 5% of that
observed in the presence of tissue. The effects of drugs used on O2
consumption are expressed as a percentage change from baseline O2
consumption. Baseline O2 consumption was measured in the cortex in
the absence and presence of L-NAME (10⫺3 mol/L) in six rats from
each group.
Effect of Agonists on O2 Consumption
Bradykinin or enalaprilat at concentrations of 10⫺7 to 10⫺4 mol/L,
or amlodipine at concentrations of 10⫺7 to 10⫺5 mol/L, were added in
a cumulative concentration-dependent manner. They were used to
measure the effects of stimulation of endogenous NO production on
renal O2 uptake. The response to these drugs was also examined after
preincubation with the NOS inhibitor L-NAME (10⫺3 mol/L) to verify
the role of NO production by NOS in the regulation of O2 uptake.
Each drug was assayed in six rats from each group in the presence or
absence of L-NAME. The effects of the superoxide dismutase mimetic
tempol (10⫺3 mol/L) were studied in an additional four rats from each
group.
Effect of NO Donor on O2 Consumption
SNAP at concentrations of 10⫺7 to 10⫺4 mol/L was added in a
cumulative concentration-dependent manner to assess the effects of
exogenous NO on renal cortical O2 uptake. The response to SNAP
Renal O2 Consumption in SHR 1789
was also examined after preincubation with L-NAME (10⫺3 mol/L).
Each condition was tested in six rats from each group.
Statistical Analyses
All data are expressed as mean ⫾ SEM. Statistical analyses of
baseline O2 consumption were performed by t test. Changes in O2
consumption caused by drug treatment were analyzed by two-way
ANOVA followed by multiple comparisons by the Tukey test (Sigma-
Stat; SPSS, Chicago, IL). Statistical significance was achieved at P ⬍
0.05.
Results
Baseline characteristics of the rats used in these studies are
listed in Table 1. Kidney/body weight, heart rate, hemoglobin
levels, and creatinine levels were similar in the two groups.
SHR rats had significantly higher systolic, diastolic, and mean
arterial pressures (P ⬍ 0.01).
Baseline Renal Cortical O2 Consumption
Baseline renal cortical O2 consumption was similar in the
two groups (WKY: 611 ⫾ 51 nmol O2/min per g, n ⫽ 6; SHR:
600 ⫾ 55 nmol O2/min per g, n ⫽ 6, P ⬎ 0.05). Addition of
the NOS inhibitor L-NAME (10⫺3 mol/L) did not significantly
alter O2 consumption (WKY: 638 ⫾ 53 nmol O2/min per g, n
⫽ 6; SHR: 634 ⫾ 66 nmol O2/min per g, n ⫽ 6, P ⬎ 0.05).
Effect of Bradykinin on Renal O2 Consumption
Cumulative doses of bradykinin (10⫺7 to 10⫺4 mol/L) pro-
duced significant, concentration-dependent decreases of renal
cortical O2 consumption in WKY and SHR rats (WKY: from
⫺2.1 ⫾ 1.1% to ⫺22.8 ⫾ 1.0%, n ⫽ 6; SHR: from ⫺0 ⫾ 0%
to ⫺13.9 ⫾ 1.9% n ⫽ 6). The depression of renal cortical O2
consumption by bradykinin was significantly less in SHR than
WKY rats at 10⫺6 to 10⫺4 mol/L of bradykinin (P ⬍ 0.05 for
each comparison) (Figure 1). Addition of L-NAME signifi-
cantly attenuated bradykinin induced decreases in O2 con-
sumption at 10⫺6 to 10⫺4 mol/L of bradykinin in WKY rats
(from ⫺0.5 ⫾ 0.5% to ⫺12.0 ⫾ 1.4%, n ⫽ 6; P ⬍ 0.05 for
each comparison), demonstrating the importance of NO syn-
thesis in the effect of bradykinin (Figure 1). In SHR rats,
addition of L-NAME had a smaller and NS effect in reversing
the action of bradykinin on renal O2 consumption (from ⫺0 ⫾
Table 1. Experimental groupsa
Parameter WKY (n ⫽ 10) SHR (n ⫽ 10)
Kidney/body weight ratio 0.34 ⫾ 0.01 0.34 ⫾ 0.01
Systolic BP (mmHg) 133.0 ⫾ 3.2 188.4 ⫾ 3.5b
Diastolic BP (mmHg) 107.9 ⫾ 2.6 147.7 ⫾ 3.2b
Mean arterial pressure (mmHg) 117.2 ⫾ 2.4 161.2 ⫾ 3.2b
Heart rate (beats/min) 402 ⫾ 16 403 ⫾ 9
Hemoglobin (g/dl) 13.1 ⫾ 0.5 13.0 ⫾ 0.3
Creatinine (mg/dl) 0.55 ⫾ 0.02 0.58 ⫾ 0.03
a
Data are mean ⫾ SEM. WKY, Wistar-Kyoto rats; SHR,
spontaneously hypertensive rats.
b
P ⬍ 0.01.
1790 Journal of the American Society of Nephrology
Figure 1. Effect of cumulative doses of bradykinin on renal cortical
O2 consumption in Wistar-Kyoto rats (WKY) (A) and spontaneously
hypertensive rats (SHR) (B) in the absence (solid circles) or presence
(open circles) of N-nitro-L-arginine methyl ester (L-NAME). Brady-
kinin caused dose-dependent decreases in O2 consumption in both
groups, although the decrease in SHR rats was significantly less than
in WKY rats. This effect was significantly attenuated by the addition
of the NOS inhibitor L-NAME (open circles) only in WKY rats. *, P
⬍ 0.05 versus WKY rats in the presence of L-NAME and SHR rats in
the absence of L-NAME.
0% to ⫺9.4 ⫾ 1.5%, n ⫽ 6, P ⬎ 0.05 for each comparison
versus no L-NAME).
Effect of Enalaprilat on Renal O2 Consumption
The angiotensin-converting enzyme inhibitor, enalaprilat
(10⫺7 to 10⫺4 mol/L), which stimulates endogenous NO pro-
duction, similarly caused concentration dependent decreases in
renal cortical O2 consumption in WKY and SHR rats (WKY:
from ⫺3.5 ⫾ 1.0% to ⫺24.1 ⫾ 2.0%, n ⫽ 6; SHR: from ⫺0.4
⫾ 0.4% to ⫺15.3 ⫾ 1.6% n ⫽ 6). The depression of cortical
O2 consumption was significantly less in SHR than WKY rats
at all dose of enalaprilat (P ⬍ 0.05 for each comparison)
(Figure 2). Addition of L-NAME significantly attenuated the
effect of enalaprilat at all doses in WKY rats (from ⫺0.0 ⫾
0.0% to ⫺12.2 ⫾ 1.6%, n ⫽ 6; P ⬍ 0.05 versus each com-
parison without L-NAME) but had no significant effect in SHR
rats (⫺0.0 ⫾ 0.0% to ⫺13.8 ⫾ 2.1% n ⫽ 6,) verifying the
J Am Soc Nephrol 13: 1788–1794, 2002
Figure 2. Effect of cumulative doses of enalaprilat on renal cortical O2
consumption in Wistar-Kyoto (WKY) (A) and spontaneously hyper-
tensive rat (SHR) (B) rats in the absence (solid circles) or presence
(open circles) of N-nitro-L-arginine methyl ester (L-NAME). Enala-
prilat caused dose-dependent decreases in O2 consumption in both
groups, although the decrease in SHR rats was significantly less than
in WKY rats at all doses of enalaprilat. This effect was significantly
attenuated by the addition of L-NAME (open circles) only in WKY
rats. *, P ⬍ 0.05 versus WKY rats in the presence of L-NAME and
SHR rats in the absence of L-NAME.
importance of NO production by NOS in the WKY, but not
SHR, rats.
Effect of Amlodipine on Renal O2 Consumption
Amlodipine (10⫺7 to 10⫺5 mol/L), which stimulates renal
NO production, significantly decreased renal cortical O2 con-
sumption in WKY and SHR rats (WKY: from ⫺4.5 ⫾ 1.4% to
⫺20.7 ⫾ 2.3%, n ⫽ 6; SHR: from ⫺1.9 ⫾ 0.6% to ⫺11.9 ⫾
0.7% n ⫽ 6). The depression of cortical O2 consumption was
significantly less in SHR than WKY rats at all doses of
amlodipine (P ⬍ 0.05 for each comparison) (Figure 3). Addi-
tion of L-NAME again significantly attenuated the effect of
amlodipine at all doses only in the WKY rats (WKY: from
⫺1.0 ⫾ 0.6% to ⫺13.0 ⫾ 1.6%, n ⫽ 6, P ⬍ 0.05 versus each
comparison without L-NAME; SHR: from ⫺0.5 ⫾ 0.5% to
⫺10.0 ⫾ 2.5% n ⫽ 6, P ⬎ 0.05).
J Am Soc Nephrol 13: 1788–1794, 2002
Figure 3. Effect of cumulative doses of amlodipine on renal cortical
O2 consumption in Wistar-Kyoto (WKY) (A) and spontaneously
hypertensive rat (SHR) (B) rats in the absence (solid circles) or
presence (open circles) of N-nitro-L-arginine methyl ester (L-NAME).
Amlodipine caused dose-dependent decreases in O2 consumption in
both groups. The decrease in SHR rats was significantly less than in
WKY rats at all doses of amlodipine. Addition of L-NAME (open
circles) significantly attenuated the effect of amlodipine only in WKY
rats. *, P ⬍ 0.05 versus WKY rats in the presence of L-NAME and
SHR rats in the absence of L-NAME.
Effect of an NO Donor (SNAP) on Renal O2
Consumption
Administration of cumulative doses of the NO donor SNAP
(10⫺7 to 10⫺4 mol/L) decreased renal cortical O2 consumption
in WKY and SHR rats to similar degrees (WKY: from ⫺0.0 ⫾
0.0% to ⫺41.7 ⫾ 1.9%, n ⫽ 6; SHR: from ⫺0.0 ⫾ 0.0% to
⫺39.7 ⫾ 1.7%, n ⫽ 6, P ⬎ 0.05 at all doses), demonstrating
no inherent difference between the strains in responsiveness of
O2 consumption to NO (Figure 4). Addition of L-NAME had
no effect on the response to SNAP (data not shown).
Effect of Oxygen Radical Scavenging with Tempol on
Renal O2 Consumption
Elevated levels of oxygen radicals have been noted in SHR
rats and have been postulated to decrease availability of NO.
Therefore, we added the superoxide dismutase mimetic tempol
(10⫺3 mol/L) to some incubations to see the effect of decreas-
Renal O2 Consumption in SHR 1791
Figure 4. Effect of cumulative doses of S-nitroso-N-acetylpenicilla-
mine (SNAP) on renal cortical O2 consumption in Wistar-Kyoto
(WKY) (A) and spontaneously hypertensive rat (SHR) (B) rats in the
absence (solid circles) or presence (open circles) of N-nitro-L-arginine
methyl ester (L-NAME). SNAP caused similar dose-dependent de-
crease in O2 consumption in both groups (P ⬎ 0.05). Addition of
L-NAME did not alter the effect of SNAP (data not shown).
ing superoxide production on the response of renal cortical O2
consumption to stimulators of NO production. Enalaprilat
again decreased O2 consumption significantly more in WKY as
compared with SHR rats (Figure 5A). Addition of tempol
significantly restored the suppression of O2 consumption by
enalaprilat (10⫺6-10⫺4 mol/L) in SHR rats to a level that was
not different from WKY rats. Suppression of O2 consumption
by amlodipine (10⫺5 mol/L) was also significantly enhanced
(Figure 5B). Tempol had no effect on the responsiveness of
WKY rats to enalaprilat or amlodipine at any dose and had no
effect on the responsiveness or either group to SNAP (Figure
5C).
Discussion
The results presented here provide evidence for the hypoth-
esis that regulation of renal oxygen consumption by NO is
impaired in SHR and that this impairment is due to ineffective
NO production as a result of NO inactivation by superoxide.
Bradykinin, enalaprilat, and amlodipine, all stimulators of NO
production, decreased renal cortical oxygen consumption in a
dose-dependent manner in WKY rats, similar to results previ-
1792 Journal of the American Society of Nephrology
Figure 5. Effect of oxygen radical scavenging with tempol on renal
cortical O2 consumption in Wistar-Kyoto (WKY) and spontaneously
hypertensive rat (SHR) rats in response to enalaprilat (A), amlodipine
(B), and S-nitroso-N-acetylpenicillamine (SNAP) (C). All drugs
caused dose-dependent decreases in O2 consumption in WKY (solid
circles) and SHR (solid triangles) rats. Addition of tempol (open
symbols) significantly decreased O2 consumption in SHR (open tri-
angles) rats at the concentrations indicated but had no effect in WKY
(open circles) rats. Suppression of O2 consumption by SNAP was
unaffected by tempol. *, P ⬍ 0.05 versus SHR in the presence of
tempol.
ously reported in normal mice and dogs (25,28). In SHR, there
was a significant decrease in the response to all three of these
stimulators of endogenous NO production, suggesting a defect
in renal NO production. The absence of a difference in basal
oxygen consumption between the two groups, despite evidence
for a defect in NO production in SHR, is most likely due to the
absence of flow in this preparation, which results in little eNOS
activation and suggests very low basal NO production. Thus,
the difference between the two groups does not become evident
until NO production is stimulated. In the presence of tempol, a
superoxide dismutase mimetic that reduces superoxide levels,
the response of oxygen consumption in SHR kidney to amlo-
dipine and enalaprilat was restored to levels seen in control
J Am Soc Nephrol 13: 1788–1794, 2002
rats. Thus, production of NO in the renal cortex in SHR
appears to be adequate to produce a similar decrease in oxygen
consumption as in controls as long as superoxide production is
inhibited. The responsiveness of kidneys from both SHR and
WKY to SNAP, and the lack of effect of tempol on the SNAP
response suggest that there is no difference in the ability of NO
to alter oxygen consumption in the two groups.
The major effect of NO on oxygen consumption occurs
through a direct action of NO on mitochondrial respiration
(22,29,30). NO binds to several enzymes in the mitochondrial
electron transport chain, including aconitase, complex I and II,
and cytochrome oxidase (reviewed in [30]). In the kidney, NO
may also decrease oxygen consumption through an effect on
sodium transport, a major determinant of oxygen usage. NO
inhibits thick ascending limb chloride flux, perhaps through an
effect on the Na⫹-K⫹-2Cl⫺ transporter, and has also been
shown to directly inhibit sodium-potassium ATPase (31–33).
Our results are consistent with and extend observations by
others on NO production and the role of superoxide on NO
availability in SHR. Both short- and long-term treatment with
tempol normalized BP and renal vascular resistance in SHR, an
effect that was shown to be dependent on NO synthesis
(19,20). Treatment with another antioxidant, lazaroid, also
lowered BP in SHR, along with decreases in the renal expres-
sion of eNOS and inducible NO synthase (21), an effect
suggested to be due to inhibition of NOS synthesis by the
newly available NO. In our studies, tempol restored NO inhi-
bition of renal oxygen consumption to normal levels. These
results all substantiate the presence of increased oxidant pro-
duction and decreased NO availability in SHR.
Our results are also interesting in the context of the increased
whole-body oxygen consumption observed in SHR (26). In
conscious dogs, inhibition of NO synthesis with nitro-L-argi-
nine leads to increased total body oxygen consumption (22).
Thus, one explanation for the changes observed in SHR is
decreased NO synthesis or availability. More recently, by using
an oxygen-sensitive ultramicroelectrode, Welch et al. (27)
demonstrated normal total renal oxygen consumption in SHR,
but a 43% reduction in tubular reabsorption of sodium, a major
determinant of renal oxygen consumption. Thus, the efficiency
of oxygen usage for sodium transport is decreased in SHR, an
effect similar to that demonstrated in conscious dogs following
inhibition of NO synthesis (24). They hypothesized that the
reduction in efficiency of oxygen usage might be due to a
relative state of NO deficiency. The data presented here sub-
stantiate that conjecture.
By regulating oxygen consumption, NO may also play an
important role in regulation of intrarenal oxygenation and
predisposition to injury. Studies by Brezis et al. (34,35) found
intrarenal pO2 of approximately 46 mmHg in the cortex and 21
to 23 mmHg in the medulla. Studies that used isolated renal
tubules have demonstrated a markedly enhanced effect of NO
on inhibition of oxygen consumption as pO2 drops to these
levels (36). Under the hypoxic conditions present in the cortex,
and even more so in the medulla, the effect of NO, or the lack
thereof, on oxygen consumption will be magnified. Inhibition
of NO synthesis with L-NAME decreases medullary pO2 (34),
J Am Soc Nephrol 13: 1788–1794, 2002
an effect postulated to be due to decreased blood flow, but also
now explained by increased oxygen consumption in the face of
low NO. L-NAME also increased hypoxic injury to tubules in
the medulla in rats, an effect reversed by infusion of the NO
donor nitroprusside (34), again an effect mediated either by
nitroprusside induced vasodilation or improved oxygenation
through inhibition of oxygen consumption by NO. Similarly, in
congestive heart failure, in which we have demonstrated de-
creased NO effect in the kidney (28), outer medullary hypoxic
damage is worsened following injection of indomethacin and
L-NAME (37).
Intrarenal pO2 is also lower in SHR than WKY rats in the
proximal and distal tubules and in the outer and deep cortex
(27), which can be explained by increased oxygen consump-
tion related to decreased NO availability. NO production in the
kidney may also be impaired in other disease states, including
diabetes, other models of hypertension, aging, and chronic
renal insufficiency (38 – 41). Thus, increased sensitivity to re-
nal insults might be expected in these states, a hypothesis
consistent with clinical observations. More importantly, evi-
dence has recently accumulated that intrarenal hypoxia may
predispose to fibrosis, leading to progressive loss of renal
function.
Hypoxia regulates the production of a broad spectrum of
growth factors, hormones, matrix components, proteases, and
protease inhibitors (reviewed in [42]). Exposure of human
proximal tubular epithelial cells to hypoxia stimulates collagen
production, decreases gelatinase A production, and increases
production of tissue inhibitor of metalloproteinase-1, changes
that would favor matrix accumulation (43). In human renal
fibroblasts, hypoxia also produces changes favoring matrix
accumulation, an effect at least partially mediated through the
hypoxia-inducible transcription factor 1 (44). Fine et al. (42)
have hypothesized that treatments such as angiotensin-convert-
ing enzyme inhibitors, which slow progression of renal disease,
may work not only by decreasing proteinuria but by increasing
microvascular flow in the interstitium and raising intrarenal
pO2. Our data suggest that angiotensin-converting enzyme
inhibitors, such as enalapril, might also increase intrarenal
oxygen tension by decreasing oxygen consumption through
stimulation of NO synthesis. Further studies of these mecha-
nisms could lead to new treatments to slow the progression of
renal disease.
Acknowledgments
This work was supported by a grant from the Westchester Artificial
Kidney Center. We acknowledge the generous support and advice of
Dr. T.H. Hintze and the secretarial assistance of Maria Feliciano.
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