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ISSN: 0886-022X (print), 1525-6049 (electronic)

Ren Fail, 2014; 36(4): 613–618

! 2014 Informa Healthcare USA, Inc. DOI: 10.3109/0886022X.2014.882240

LABORATORY STUDY

Circulating purine compounds, uric acid, and xanthine oxidase/

dehydrogenase relationship in essential hypertension and end
stage renal disease
Milojkovic Boban1, Gordana Kocic1, Sonja Radenkovic2, Radmila Pavlovic3, Tatjana Cvetkovic1, Marina Deljanin-Ilic4,
Stevan Ilic4, Milojkovic D. Bobana5, Boris Djindjic2, Dijana Stojanovic2, Dusan Sokolovic1, and
Tatjana Jevtovic-Stoimenov1
1
Department of Biochemistry, 2Department of Pathophysiology, 3Department of Chemistry, Medical Faculty University Nis, Nis, Serbia, 4Institute for
Cardiology Niska banja Medical Faculty, Nis, Serbia, and 5Clinic for Surgery Clinical Center University Nis, Nis, Serbia
Ren Fail Downloaded from informahealthcare.com by Nyu Medical Center on 02/10/15

Abstract Keywords
Purine nucleotide liberation and their metabolic rate of interconversion may be important in ADP, AMP, ATP, circulating purine
the development of hypertension and its renal consequences. In the present study, blood nucleotides, dialysis, essential
triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) hypertension, renal failure, xanthine
breakdown pathway was evaluated in relation to uric acid concentration and xanthine dehydrogenase, xanthine oxidase
dehydrogenase/xanthine oxidase (XDH/XO) in patients with essential hypertension, patients
with chronic renal diseases on dialysis, and control individuals. The pattern of nucleotide History
catabolism was significantly shifted toward catabolic compounds, including ADP, AMP, and uric
For personal use only.

acid in patients on dialysis program. A significant fall of ATP was more expressed in a group of Received 23 October 2013
patients on dialysis program, compared with the control value (p50.001), while ADP and AMP Revised 24 November 2013
were significantly increased in both groups of patients compared with control healthy Accepted 4 December 2013
individuals (p50.001), together with their final degradation product, uric acid (p50.001). The Published online 6 February 2014
index of ATP/ADP and ATP/uric acid showed gradual significant fall in both the groups,
compared with the control value (p50.001), near five times in a group on dialysis. Total XOD
was up-regulated significantly in a group with essential hypertension, more than in a group on
dialysis. The activity of XO, which dominantly contributes reactive oxygen species (ROS)
production, significantly increased in dialysis group, more than in a group with essential
hypertension. In conclusion, the examination of the role of circulating purine nucleotides and
uric acid in pathogenesis of hypertension and possible development of renal disease, together
with XO role in ROS production, may help in modulating their liberation and ROS production in
slowing progression from hypertension to renal failure.

Introduction Adenine nucleotides can be found in circulation, after their

release from peripheral tissues, endothelial cells, stimulated
It was documented that renal failure, followed by the end-stage
renal disease (ESRD) would be primarily caused by hyper-
blood platelets, immune cells, and during stimulation of 14
20
sympathetic adrenergic and purinergic nerves.12–16 Tissue
tension in about of 30% cases.1–5 Despite the fact that essential
damage, necrosis, metabolic stress, hypoxia, endothelial
hypertension may occurs in the absence of any known cause, a
ischemia during shear stress, myocardial ischemic attack,
number of metabolic disturbances, including hyperuricemia,
muscle exercise, or smooth muscle cells contraction, may
obesity as well as hyperactive sympathetic nervous system
contribute to their liberation into extracellular space. Their
may be responsible for pathophysiological and hemodynamic
release may occur via exocytosis or diffusion across damaged
features.6,7 Hyperuricemia may induce renal failure through
membrane. From the other side, hyperactive sympathetic
the crystal-dependent and crystal-independent mechanisms,
nervous system, or adrenal medulla stimulation may produce
by inducing endothelial dysfunction, renal vasoconstriction,
an increase in adenine nucleotide levels and platelet activation
and activation of the renin–angiotensin system.8–11 Purine
(aggregation).17–20 Once present in the circulation, they may
nucleotide degradation pathway may be important in the
exert vasoactive, immunomodulatory, and prothrombotic
development of hypertension and its renal consequences.
responses. Very strong vasoactive properties of different
purine nucleotides mainly depend on their phosphate form,
whether they are in triphosphate, diphosphate, and monopho-
Address correspondence to Professor Gordana Kocic, MD, MSc, PhD,
Department of Biochemistry, Medical Faculty University Nis, Bul Dr sphate forms.18,19 Their vasoactive effects occur mainly via
Zorana Djindjica 81, 18000 Nis, Serbia. E-mail: kocicrg@yahoo.co.uk purinergic system.20–23 Different pharmacologic drugs
 614 M. Boban et al.
(like acetyl-choline or bradykinin) may also lead to purine
nucleotide liberation.24 Primary disorders of purine synthesis
or salvage pathway may increase uric acid metabolism,
leading to gout and renal calculi. Secondary hyperuricemia
may be frequently present in anemia, central nervous system
dysfunction, obesity or diabetes, commonly contributing to
the development of serious renal failure.9,25
Human enzyme xanthine oxidoreductase (XOD) is a rate-
limiting enzyme involved in terminal catabolic pathway of
purine bases, adenine and guanine, through hypoxanthine and
xanthine to uric acid. In tissues and circulation, it can exist in
two interconvertible forms: xanthine dehydrogenase (XDH)
and xanthine oxidase (XO). The interconversion from XDH
to XOD can be stimulated by the oxidation of the pre-
sent sulfhydryl residues or by stimulated proteolysis.25,26
Since XO reaction, simultaneously with uric acid, produces
reactive oxygen species (ROS), this enzyme represents a main
source of ROS liberation in circulation. XO activity in
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circulation is mainly elevated in ischemic–reperfusion condi-
tions, coronary disease, endothelial dysfunction, inflamma-
tory conditions followed by cytokine liberation (IL-1, IL-6,
and TNF-a), and bacterial lipopolysaccharide presence, as
well as following steroid hormone treatment.25–30
Since the excess accumulation of different purine nucleo-
tides, together with increased XOD expression and XO
activity, can be regarded as an independent pathogenic factor
for endothelial damage, alteration of metabolic processes
For personal use only.
in the kidneys and pro-coagulant phase, in the present study
the adenine nucleotide breakdown pathway in blood was
evaluated in relation to uric acid concentration and XDH/XO
ratio in patients with essential hypertension and patients with
chronic renal diseases on dialysis.
Patients and methods
Three groups of patients were included in the study: I,
patients with essential hypertension without renal pathology
(30; 21/9 females/males); II, patients with chronic renal
diseases on dialysis (28; 16/12 females/males); and III,
controls healthy persons (20; 12/8 females/males). Control
group was allocated from randomly selected, healthy indi-
viduals, blood donor volunteers, and age-matched. All
participants gave written informed consent for analysis.
Patients with essential hypertension and the end stages of
chronic kidney diseases on dialysis were recruited from the
Clinic for Nephrology and Hemodialysis Medical Faculty,
University of Nis. The diagnosis of disease was based on a
survey, which included standard clinical and imaging methods
and laboratory examination of blood and urine. Routine
biochemical methods were used to determine serum concen-
trations of creatinine, urea, and other clinically relevant
parameters for renal function in establishing a diagnosis of
stages of chronic kidney disease. They were selected accord-
ing to the National Kidney Foundation (NKF) criteria.31
Patients were checked for history of previous or present
infections as well. None of them had the above-mentioned
exclusion criteria.
Normal blood pressure was defined as a systolic blood
pressure of less than 130 mmHg and a diastolic blood
pressure of less than 80 mmHg, while systolic pressures of
Ren Fail, 2014; 36(4): 613–618
130–139 mmHg and diastolic pressures of 80–89 mmHg were
considered as borderline hypertension. Hypertension was
defined as a systolic blood pressure of at least 140 mmHg,
a diastolic blood pressure of at least 90 mmHg, or both,
measured in a sitting position, repeated at least three times,
according to the World Health Organization (WHO)
classification.6,32
Blood collection was done from cubital vein and patients
and control group were advised not to perform any intensive
physical training or to eat meat at least 2 d before the analysis.
For each participant, a 10 mL blood sample was collected
after a 30-min sitting period. Routine biochemical analyses
were determined on separated plasma on Automatic analyzer
A24 for In Vitro Diagnostics (manufactured by Biosystems
SA, Barcelona, Spain). For the evaluation of purine nucleo-
tides concentration, fresh blood was immediately processed
for their isolation and the content of intermediates of purine
metabolism was determined in the whole by HPLC analysis.33
XOD and XO were measured in plasma according to the
liberation of uric acid by using xanthine as substrate,34 in the
presence of NADH (for XOD) or absence of NADH (for XO)
when only molecular oxygen was electron acceptor.28
The XDH activity was calculated by subtracting from XOD
the XO activity.
Results
The results of clinical and biochemical parameters are shown
in Table 1. Increased creatinine and urea, together with
glomerular filtration rate (GFR) fall, were expected to be
present in a group on dialysis. Hypertension was confirmed in
both clinical groups.
Figure 1 represents the concentration of circulating blood
nucleotides, triphosphate (ATP), adenosine diphosphate
(ADP), and adenosine monophosphate (AMP) in investigated
groups. In the fresh whole blood samples, the ATP, ADP,
AMP, and uric were detected as circulating purine com-
pounds. A significant fall in the level of ATP was documented
in both clinical groups of patients, more in a group of patients
on dialysis program, compared with control value (p50.001).
Both degradation nucleoside phosphate products, ADP and
AMP, were significantly increased in both groups of patients
compared with control healthy individuals (p50.001),
together with their final degradation product, uric acid
(p50.001). Since simultaneous purine nucleotide degradation
may consequently lead to the production of uric acid, the
index of ATP/ADP and ATP/uric acid was evaluated and it is
shown in Figure 2. Gradual significant fall in both indexes
was observed in both groups, compared with the control value
(p50.001), which was almost five times in a group on
dialysis. Significant difference was observed between two
clinical groups, where group with essential hypertension
had almost two times higher index of ATP/ADP than group
on dialysis and also higher index ATP/uric acid. Figure 3
represents the activity of total XOD activity and both XO and
XDH activities in plasma of the investigated groups. Total
XOD activity was the most significantly expressed in a group
with essential hypertension, more than in a group on dialysis.
But this group still remained higher XDH activity, signifi-
cantly more than control or dialysis group. From the other
 DOI: 10.3109/0886022X.2014.882240 Circulating purine compounds, uric acid, and XDH/XO relationship 615

side, the activity of XO, which dominantly contributes to
uric acid and ROS production, was significantly increased
in dialysis group, more than in a group with essential
hypertension.
Discussion
Obtained results from our study showed disturbed circulating
blood purine nucleotide level, which exerted strong shift
toward the increased degradation products, such as ADP and
AMP in relation to ATP form. The concentration of their
terminal degradation product-uric acid reflected this condi-
tion as well (Figure 1) what was expressed on evaluated
indexes, ATP/ADP, and ATP/ uric acid (Figure 2). The
activity of plasma XDH/XO exerted strong shift toward
XO activity, especially in a dialysis group (Figure 3).
Follow-up studies before effective antihypertensive ther-
apy documented that between one-third and two-thirds of
subjects with essential hypertension may develop proteinuria,
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ligand-gated P2X receptors. Since vascular tone in blood
vessels can be controlled by perivascular nerves and endo-
thelial cells, released ATP may exert dual function, depending
on the source of its liberation. If being released as a
neurotransmitter by sympathetic nerves, it can act via P2X
receptors of vascular smooth muscle cells, by producing
vasoconstriction. From the other side, the ATP released from
endothelial cells by shear stress, ischemia, or hypoxia may be
recognized by P2Y receptors on endothelial cells, simultan-
eously releasing nitric oxide, which produces vasodilata-
tion.20–23 In the healthy conditions, when vessel wall and
endothelium are intact, the main effect of ATP is vasodilatory,
presumably due to nitric oxide release. During atherosclerosis
or pro-thrombotic states, the main source of ATP is
aggregated thrombocytes. Since released ATP may be able
to act through damaged endothelium on vascular smooth
muscle cells, the effect is usually vasoconstrictory. Enhanced
release of ATP, beside hypoxia, may be provoked by some

with one-third developing renal insufficiency and ESRD,
about 10% dying from uremia.1–3,35–37 Extracellular purine
nucleotides, ATP, ADP and AMP represent potent vasoactive
and prothrombotic compounds of the vascular wall, by
triggering signaling via G-protein-coupled P2Y and via
Table 1. Plasma urea, creatinine, and blood pressure in the investigated
groups.
For personal use only.
pharmacologic agents, like calcium agonists, pro-thrombotic
compounds (thrombin), and bacterial toxins (lipopolysacchar-
ide).16–20 The ATP degradation products, such as AMP and
uric acid, may mediate vasoconstrictory effect via stimulation
of angiotensin system and may contribute to damage of the
endothelial barrier.9–11
Metabolic deterioration of the macro energetic ATP
compound in cells may reflect on current changes in blood
purine nucleotides. The relative amounts of the three adenine

Investigated Essential nucleotides (adenylate pool) may summarize the energy status
parameters Control20 hypertension30 Dialysis28 of a cell, known as the adenylate energy charge (ATP + ADP)/
Plasma U urea 4.09 ± 1.35 6.06 ± 2.08 22.48 ± 7.43*** ooo (ATP + ADP + AMP). Concerning the ATP degradation, the
(mmol/L) ATP/ADP ratio and the ATP/AMP ratio should be physiolo-
Plasma creatinine 72.06 ± 5.18 84.37 ± 13.97 597.03 ± 101.88*** ooo gically maintained as high as possible in order to perform a
(mmol/L) variety of energy-dependent cellular activities, such as muscle
Systolic TA 134.45 ± 12.44 163.18 ± 24.56 150.34 ± 23.28
(mmHg) contraction, motility, and ion pumping. If the ratio physiolo-
Dyastolic TA 79.98 ± 9.99 94.65 ± 17.36 89.99 ± 20.98 gically tended to rise above 0.7, it may reflect potential for
(mmHg) high energy phosphoryl transfer, while fall in this charge
***p50.001, statistical significance compared with the control. represents the cell energy crisis state.38 During shear stress
ooo
p50.001, statistical significance compared with the patients with and ischemia, the extracellular ATP concentration may
essential hypertension. increase up to 10-fold, but the adenine nucleotides are

Figure 1. The concentration of circulating 1200 µmol/L

blood nucleotides, ATP, ADP, AMP, and uric
acid in investigated groups. For the evalu- 1000
ation of purine nucleotides concentration, ooo
fresh blood was immediately processed for
their isolation after adding stabilizing solu- 800 ***
ooo
tion of retard ATP degradation. ***p50.001 *** ***
compared with the control; *p50.05 com- 600 ***
pared with the control; ooop50.001 com-
pared with the essential hypertension group. 400 *
200 *** ***
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 616 M. Boban et al. Ren Fail, 2014; 36(4): 613–618

Figure 2. The index of ATP/ADP and ATP/ 6

uric acid in investigated groups. For the
evaluation of purine nucleotides concentra- 5
tion, fresh blood was immediately processed
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Ren Fail Downloaded from informahealthcare.com by Nyu Medical Center on 02/10/15

Figure 3. The activities of total Xanthine 25

oxidoreductase (XOD), xanthine oxidase U/L
(XO), and xanthine dehydrogenase (XDH) in ***
plasma of investigated groups. Xanthine 20 *
oxidoreductase (XOD) and xanthine oxidase
(XOD) were measured in plasma according to
the liberation of uric acid,34 in the presence 15
of NADH (XOD) or absence of NADH (XO) *
For personal use only.

when only molecular oxygen was electron

acceptor. ***p50.001 compared with the 10 *** ***
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sequentially dephosphorylated by membrane-bound ecto-
nucleotidases and circulating purine catabolic degrading
enzymes. ADP represents one of the major contributors to
the thrombotic effects, since ADP antagonists act as potential
anti-thrombotic agents.14,24 The results obtained in our study
documented significant fall in whole blood ATP level with
consequent production of ADP, AMP, and uric acid (Figures 1
and 2). Concentration of ADP was documented to be
significantly (about 50%) higher in patients with essential
hypertension, but almost three times higher in patients on
dialysis (Figure 1). Increased concentration of uric acid in this
way may be result of consequent final degradation of purine
compounds. The impairment of fractional excretion of uric
acid was also observed in patients with essential hyperten-
sion.39,40 It may explain their role in augmenting not only
hypertension but also platelet activity contributing to
atherosclerosis acceleration and development of renal com-
plications.10,11 Recent studies documented that extracellular
purine nucleotides and uric acid may represent important
extrarenal stimuli for the development of hypertension
and ESRD, due to diverse metabolic functions and due to
participation in the regulation of renal function, rennin–
angiotensin system, vascular tone, and coagulation cas-
cade.9,40 Since their concentration was especially disturbed
in patients on dialysis, our results may suggest their contri-
bution to the development and progression of chronic kidney
disease. The harmful effect of hyperuricemia was confirmed,
by reducing uric acid level, what may slow down renal
progression in subjects with hypertension. Results obtained in
experimental hyperuricemia have shown that the development
of the preglomerular arteriolar lesions resulted in hemo-
dynamic changes followed by reduced renal blood flow and
 DOI: 10.3109/0886022X.2014.882240 Circulating purine compounds, uric acid, and XDH/XO relationship
glomerular hypertension. High level of uric acid may activate
angiotensin II, may potentiate renal vasoconstriction, and may
up-regulate angiotensin type 1 receptors on vascular smooth
muscle cells, stimulating in this way, the renin–angiotensin
system.9–11,40 The possible causal relationship of blood
pressure with the uric acid level was documented by using
xanthine oxidase inhibitors or uricosuric agents (allopurinol
or benziodarone) to control essential hypertension.41
Hominoid evolution of primates led to uricase mutation and
deletion, where uric acid appeared as the terminal purine
degradation product. The hypothesis was that the uricase
mutation helped to maintain uric acid high enough to
maintain blood pressure acutely (via stimulation of the
renin–angiotensin system) and chronically (by inducing salt-
sensitivity) in order to develop and maintain the human
upright posture and locomotion.42,43 During experimental
hyperuricemia, the renal interstitial inflammation and tubular
injury were documented to induce renal microvascular and
Ren Fail Downloaded from informahealthcare.com by Nyu Medical Center on 02/10/15
interstitial disease. Uric acid is able to stimulate vascular
smooth muscle cell proliferation through the activation of the
mitogen-activated protein (MAP) kinase, extracellular signal-
regulated kinase (Erk1/2), stimulation of platelet-derived
growth factor (PDGF), cyclooxygenase-2 (COX-2) expres-
sion, and production of thromboxane.44–47
The results about plasma XOD activity and its components
XDH and XO are shown in Figure 3. Total XOD activity
showed up-regulated activity in a group with essential
For personal use only.
hypertension, more than in a group on dialysis. It may
suggest that some endothelial factors may contribute to the
increased expression of enzyme, since this group still
remained higher XDH activity, significantly more than
control or dialysis group. From the other side, the activity
of XO, which dominantly contributes to uric acid and ROS
production, significantly increased in a dialysis group, more
than in a group with essential hypertension. It was docu-
mented that circulating XO may bind to glycosaminoglycans
on the surface of endothelial cells, which contribute to its
activation and longer stability.25,26,29 Taking into consider-
ation both XO-catalytic reaction products, uric acid and ROS,
our results are very close to the role of angiotensin and
oxidative stress in essential hypertension what was commonly
documented.9–11 The role of ROS in the development of
renovascular hypertension was documented as well and our
results pointed out the role of XO in their generation.48
Deposited XO via ROS generation may directly damage
endothelium, contributing further to renal damage.
Conclusion
By simultaneously investigating circulating blood nucleotide
level in patients with essential hypertension and patients on
dialysis program, we showed that the pattern of nucleotide
catabolism was significantly shifted toward catabolic com-
pounds, including ADP, AMP, and uric acid in patients on
dialysis program. Further, we observed significant increase in
plasma XO activity, mainly due to the interconversion
between dehydrogenase form (XDH) in patients on dialysis,
but due to general up-regulation of total XOD activity, mainly
in group with essential hypertension. The examination of the
role of circulating purine nucleotides and uric acid in
617
pathogenesis of hypertension and possible development of
renal disease, together with XO role, may help in modulating
their liberation and ROS production in slowing progression
from hypertension to renal failure.
Declaration of interest
The authors report no conflicts of interest. The authors alone
are responsible for the content and writing of this article.
The work is supported by the Ministry of Science Serbia
TR31060.
References
1. Rostand SG. Is renal failure caused by primary hypertension?
Why does the controversy continue? Nephrol Dial Transpl. 1998;
13:3007–3010.
2. Bidani AK, Griffin KA. Pathophysiology of hypertensive renal
damage. Implications for therapy. Hypertension. 2004;44:595–601.
3. Madhaven S, Stockwell D, Cohen H, Alderman MH. Renal function
during antihypertensive treatment. Lancet. 1995;345:749–751.
4. Rostand SG, Brown G, Kirk K, Rutsky EA, Dustan HP. Renal
insufficiency in treated essential hypertension. N Engl J Med. 1989;
320:684–688.
5. Brazy PC, Fitzwilliam JF. Progressive renal disease: role of race
and antihypertensive medications. Kidney Int. 1990;37:1113–1119.
6. Kanellis J, Nakagawa T, Herrera-Acosta J, Schreiner GF,
Rodriguez-Iturbe B, Johnson RJ. A single pathway for the
development of essential hypertension. Cardiol Rev. 2003;11:
180–196.
7. Fields LE, Burt VL, Cutler JA, Hughes J, Roccella EJ, Sorlie P.
The burden of adult hypertension in the United States 1999 to 2000.
A rising tide. Hypertension. 2004;44:398–404.
8. Nakagawa T, Mazzali M, Kang DH, et al. Hyperuricemia causes
glomerular hypertrophy in the rat. Am J Nephrol. 2003;23:2–7.
9. Johnson R, Segal MS, Srinivas T, et al. Essential hypertension,
progressive renal disease, and uric acid: a pathogenetic link? JASN.
2005;16(7):1909–1919.
10. Mazzali M, Hughes J, Kim YG, et al. Elevated uric acid increases
blood pressure in the rat by a novel crystal-independent mechanism.
Hypertension. 2001;38:1101–1106.
11. Sanchez-Lozada LG, Tapia E, Santamaria J, et al. Mild hyperur-
icemia induces vasoconstriction and maintains glomerular hyper-
tension in normal and remnant kidney rats. Kidney Int. 2005;67:
237–247.
12. Pearson JD, Gordon JL. Vascular endothelial and smooth muscle
cells in culture selectively release adenine nucleotides. Nature.
1979;281:384–386.
13. Gordon JL. Extracellular ATP: effects, sources and fate. Biochem J.
1986;233:309–319.
14. Borst M, Schrader J. Adenine nucleotide release from isolated
perfused guinea pig hearts and extracellular formation of adenosine.
Circ Res. 1991;68:797–806.
15. Coade S, Pearson J. Metabolism of adenine nucleotides in human
blood. Circ Res. 1989;65:531–537.
16. Vial C, Owen P, Opie LH, Posel D. Significance of release of
adenosine triphosphate and adenosine induced by hypoxia or
adrenaline in perfused rat heart. J Mol Cell Cardiol. 1987;19:
187–197.
17. Bergfeld GR, Forrester T. Release of ATP from human erythrocytes
in response to a brief period of hypoxia and hypercapnia.
Cardiovasc Res. 1992;26:40–47.
18. Bodin P, Burnstock G. ATP-stimulated release of ATP by human
endothelial cells. J Cardiovasc Pharmacol. 1996;27:872–875.
19. Beigi R, Kobatake E, Aizawa M, Dubyak GR. Detection of local
ATP release from activated platelets using cell surface-attached
firefly luciferase. Am J Physiol. 1999;276:C267–C278.
20. Bodin P, Burnstock G. Increased release of ATP from endothelial
cells during acute inflammation. Inflamm Res. 1998;47:351–354.
21. Schwiebert LM, Rice WC, Kudlow BA, Taylor AL, Schwiebert
EM. Extracellular ATP signaling and P2X nucleotide receptors in
monolayers of primary human vascular endothelial cells. Am J
Physiol. 2002;282:C289–C301.
 618 M. Boban et al.
22. North RA. Molecular physiology of P2X receptors. Physiol Rev.
2002;82:1013–1067.
23. Ralevic V, Burnstock G. Receptors for purines and pyrimidines.
Pharmacol Rev. 1998;50:413–492.
24. Darius H, Stahl GL, Lefer AM. Pharmacological modulation of
ATP release from isolated rat hearts in response to vasoconstrictor
stimuli using a continuous flow technique. J Pharmacol Exp Ther.
1987;240:542–547.
25. Berry CE, Hare JM. Xanthine oxidoreductase and cardiovascular
disease: molecular mechanisms and pathophysiological implica-
tions. J Physiol. 2004;16;555(Pt 3):589–606.
26. Pacher P, Nivorozhkin A, Szabo C. Therapeutic effects of xanthine
oxidase inhibitors: renaissance half a century after the discovery of
allopurinol. Pharmacol Rev. 2006;58(1):87–114.
27. Zhang Z, Blake DR, Stevens CR, et al. A reappraisal of xanthine
dehydrogenase and oxidase in hypoxic reperfusion injury: the role
of NADH as an electron donor. Free Radic Res. 1998;
28(2):151–164.
28. Spiekermann S, Landmesser U, Dikalov S, et al. Electron spin
resonance characterization of vascular xanthine and NAD(P)H
oxidase activity in patients with coronary artery disease: relation to
endothelium-dependent vasodilation. Circulation. 2003;
107(10):1383–1389.
Ren Fail Downloaded from informahealthcare.com by Nyu Medical Center on 02/10/15
29. de Jong JW, Schoemaker RG, de Jonge R, et al. Enhanced
expression and activity of xanthine oxidoreductase in the failing
heart. J Mol Cell Cardiol. 2000;32(11):2083–2089.
30. Pfeffer KD, Huecksteadt TP, Hoidal JR. Xanthine dehydrogenase
and xanthine oxidase activity and gene expression in renal epithelial
cells. Cytokine and steroid regulation. J Immunol. 1994;
153(4):1789–1797.
31. National Kidney Foundation—K/DOQI. Clinical practice guide-
lines for chronic kidney disease: evaluation, classification and
stratification. Am J Kidney Dis. 2002;39(suppl. 1):S1–S266.
32. Joint National Committee on Prevention, Detection, Evaluation,
For personal use only.
and Treatment of High Blood Pressure. The sixth report of the Joint
National Committee on prevention, detection, evaluation, and
treatment of high blood pressure (JNC VI). Arch Intern Med. 1997;
157:2413–2446.
33. Coolen E, Arts I, Swennen E, Bast A, Cohen Stuart M, Dagnelie P.
Simultaneous determination of adenosine triphosphate and its
metabolites in human whole blood by RP-HPLC and UV-detection.
J Chromatogr B. 2008;64:43–51.
34. Kizaki H, Sakurada T. Simple micro-assay methods for enzymes of
purine metabolism. J Lab Clin Med. 1977;89(5):1135–1144.
Ren Fail, 2014; 36(4): 613–618
35. Perneger TV, Nieto J, Whelton PK, Klag MJ, Comstock GW, Szklo
M. A prospective study of blood pressure and serum creatinine.
Results from the ‘‘Clue’’ study and the ARIC study. JAMA. 1993;
269:488–493.
36. Haroun MK, Jaar BG, Hoffman SC, Comstock GW, Klag MJ,
Coresh J. Risk factors for chronic kidney disease: a prospective
study of 25,534 men and women in Washington County, Maryland.
J Am Soc Nephrol. 2003;14:2394–2441.
37. Lindeman RD, Tobin JD, Shock NW. Hypertension and the kidney.
Nephron. 1989;47(Suppl. 1):62–67.
38. Goldbeter A. Modulation of the adenylate energy charge by
sustained metabolic oscillations. FEBS Lett. 1974;43(3):327–330.
39. Tykarski A, Głuszek J, Banaszak F. Value of oxypurines and uric
acid in plasma, renal excretion of oxypurines and uric acid as well
as plasma adenosine deaminase and AMP deaminase activity in
patients with essential hypertension. Pol Arch Med Wewn. 1993;
89(3):223–229.
40. Kinugawa T, Ogino K, Osaki S, et al. Altered purine nucleotide
degradation during exercise in patients with essential hypertension.
Metabolism. 2001;50(6):646–650.
41. Romero JC, Reckelhoff JF. State-of-the-Art lecture: role of
angiotensin and oxidative stress in essential hypertension.
Hypertension. 1999;34:943–949.
42. Watanabe S, Kang DH, Feng L, et al. Uric acid, hominoid
evolution, and the pathogenesis of salt-sensitivity. Hypertension.
2002;40:355–360.
43. Johnson RJ, Srinivas T, Cade JR, Rideout BA, Oliver WJ. Uric
acid, evolution and primitive cultures. Semin Nephrol. 2005;25:3–8.
44. Graham A, McLees A, Kennedy C, Gould GW, Plevin R.
Stimulation by the nucleotides, ATP and UTP of mitogen-activated
protein kinase in EAhy 926 endothelial cells. Br J Pharmacol.
1996;117:1341–1347.
45. Kolosova IA, Mirzapoiazova T, Adyshev D, et al. Signaling
pathways involved in adenosine triphosphate-induced endothelial
cell barrier enhancement. Circ Res. 2005;97:115–124.
46. Patel V, Brown C, Goodwin A, Wilkie N, Boarder MR.
Phosphorylation and activation of p42 and p44 mitogen-activated
protein kinase are required for the P2 purinoceptor stimulation of
endothelial prostacyclin production. Biochem J. 1996;320:221–226.
47. Bogatcheva NV, Dudek SM, Garcia JG, Verin AD. Mitogen-
activated protein kinases in endothelial pathophysiology. J Investig
Med. 2003;51:341–352.
48. Lerman LO, Nath KA, Rodriguez-Porcel M, et al. Increased
oxidative stress in experimental renovascular hypertension.
Hypertension. 2001;37:541–546.