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In Vivo Studies
of a cluster of cardiovascular
disease risk factors that include obesity, glucose intolerance,
hyperinsulinemia, dyslipidemia, and hypertension (32). The
prevalence of metabolic syndrome is increasing and now affects 27% of the population in the United States (11). The
epidemic correlates with pronounced changes in the environment, behavior, and lifestyle and is considered one of the main
threats to human health worldwide. Metabolic syndrome confers a greater than threefold increased risk for cardiovascular
Experiment I: treatment of fructose-induced hyperuricemia with allopurinol. Male Sprague-Dawley rats (150 200 g) were housed in
standard conditions and fed a control (n 7) or 60% fructose diet
(Harlan, Madison, WI, n 14) for 10 wk. The control diet contained
46% carbohydrate, which is mainly composed of starch, whereas the
fructose diet contained 60% fructose as the carbohydrate. The caloric
content of these diets are 3.1 and 3.6 kcal/g, respectively. At 4 wk,
blood samples were obtained at 11 AM after 4 h of fasting. One-half
of the fructose-fed rats were administered allopurinol (150 mg/l in the
drinking water, Sigma, St. Louis, MO) for an additional 6 wk to lower
serum uric acid. Fresh drinking water containing allopurinol was
replaced every 2 days. Rats were divided into three groups: control,
fructose, and fructoseallopurinol. At 10 wk, an oral glucose tolerance test (OGTT) was performed, in which rats were fasted overnight
(16 h) and then administered 1.5 g/kg of a 50% glucose solution by
gavage. Blood was sampled at 0, 30, 60, and 120 min for blood
glucose and serum insulin measurement. The rats were then killed.
Deceased.
Address for reprint requests and other correspondence: T. Nakagawa, Div.
of Nephrology, Hypertension, and Transplantation, PO Box 100224, Univ. of
Florida, Gainesville, FL 32610 (e-mail: nakagt@medicine.ufl.edu).
The costs of publication of this article were defrayed in part by the payment
of page charges. The article must therefore be hereby marked advertisement
in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
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Statistical Analysis
All values presented are expressed as means SD and analyzed by
one-way ANOVA or by unpaired Students t-test. Significance was
defined as P 0.05.
RESULTS
In Vivo Studies
Serum uric acid levels, systolic blood pressure, and fasting
insulin levels were elevated in fructose-fed rats compared with
rats fed a control diet at 4 wk (Table 1). In addition, the body
weight of fructose-fed rats tended to increase compared with
rats fed a normal diet (Table 1). These data demonstrate that
fructose feeding induces early features of metabolic syndrome
in rats.
To examine the role of uric acid in this model, one-half of
the fructose-fed rats were treated with allopurinol (a xanthine
oxidase inhibitor) for 6 additional wk. This treatment was
effective at lowering uric acid, whereas the fructose-fed rats
that did not receive treatment continued to be hyperuricemic
(Fig. 1A). In addition, we examined the urinary excretion of
uric acid in these animals to clarify the mechanisms of hyperuricemia in fructose-fed rats. As shown in Fig. 1B, fructose-fed
rats had a lower urinary excretion of uric acid. Interestingly,
allopurinol prevented the reduced excretion of uric acid in
fructose-fed rats.
Fructose-fed rats treated with allopurinol showed an improvement in metabolic syndrome. Allopurinol significantly
reduced systolic blood pressure in fructose-fed rats (Fig. 1C),
although pressures remained higher than that observed in
control rats. Fructose-fed rats also developed marked hypertriglyceridemia that was abolished by allopurinol treatment
(Fig. 1D). The reduction in serum uric acid correlated
directly with the decrease in triglyceride levels (Fig. 1E).
Fructose-fed rats also showed an increase in body weight
compared with controls. Allopurinol prevented the increase
in body weight, although this did not reach significance
(522 57 g in fructose vs. 470 28 g in control and 474
37 g in fructoseallopurinol, P not significant).
While no groups developed fasting or postprandial hyperglycemia (Fig. 2A), fructose-fed rats developed fasting hyperinsulinemia that was reversed with allopurinol (Fig. 2B). Postprandial hyperinsulinemia also occurred in fructose-fed rats
administered an OGTT, and this was partially but significantly
lower in allopurinol-treated rats (Fig. 2B), resulting in improved insulin sensitivity (Fig. 2C).
We also examined the effectiveness of allopurinol in preventing the development of metabolic syndrome, as opposed to
Control (n 7)
Fructose (n 14)
19012
35715
1273
1.30.3
12164
1880.1
37522
14815
2.40.3
17651
NS
P0.05
P0.05
P0.01
P0.05
Values are means SD. n, No. of rats; BP, blood pressure; NS, not
significant.
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may be a mechanism for the development of insulin resistance (36). Previously, uric acid has been shown to potently
reduce NO levels in cultured bovine endothelial cells (18).
To further examine this relationship, we examined the acute
effect of uric acid on acetylcholine-induced vasodilation of
rat AA rings. As shown in Fig. 4, uric acid dose dependently
blocked the vasorelaxation of AA rings in response to
acetylcholine.
DISCUSSION
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Control
(n 8)
Allopurinol
(n 8)
Benzbromarone
(n 8)
1627
46969
145268
1.50.5
1643
50437
1562118
0.50.4
1608
46843
149490
1.10.3
60% Dextrose
(n 8)
60% Fructose
(n 8)
60% Fructose
Benzbromarone
(n 8)
1447
35314
70010
1.40.3
11228
11243
1445
36417
7096
2.10.9*
41960
20462
1447
36020
69815
1.10.4
29386
14742
Values are means SD. n, No. of rats. *P 0.05 vs. dextrose. P 0.01
vs. fructose. P 0.01 vs. dextrose. P 0.05 vs. fructose.
AJP-Renal Physiol VOL
studies suggest that uric acid may also have a contributory role
in the development of insulin resistance.
Hypertriglyceridemia was completely blocked by lowering
of uric acid with allopurinol in this study. Compatible with our
findings, it has been shown that the association of elevated
serum uric acid with hypertriglyceridemia is stronger than with
insulin sensitivity (41). Interestingly, treatment of hypertriglyceridemia with finofibrate or atorvastatin also reduces serum
uric acid (1, 12, 27). Uricosuric agents such as benziodarone
also lower serum triglycerides (12). Although the role of uric
acid in the metabolism of triglycerides remains unknown, uric
acid might be involved in either the overproduction or the
reduction of clearance of triglycerides. A decrease in the
clearance of triglycerides in fructose-fed rats has been attributed to a reduction in lipoprotein lipase activity in endothelial
cells (19, 30); whether this is mediated by uric acid remains to
be determined. An alternative explanation is the possibility that
the de novo increase in purine synthesis observed in fructosefed rats may be pathogenetically linked to hepatic fatty acid
synthesis, resulting in overproduction of triglycerides (25, 41).
Endothelial dysfunction is a hallmark of metabolic syndrome (7). Therefore, we investigated the role of uric acid in
endothelial dysfunction as a mechanism for insulin resistance.
We showed that uric acid dose dependently blocked acetylcholine-mediated arterial dilation (Fig. 4), suggesting that uric acid
can impair endothelial function. In addition, we have found
that uric acid potently reduces endothelial NO bioavailability
in both cell culture and in experimental animal models (18). In
turn, reducing endothelial NO levels is a known mechanism for
inducing insulin resistance (33). Thus endothelial NOS-deficient mice exhibit the features of metabolic syndrome (5). The
mechanism is due to a blockade of insulin action, as insulin
stimulates glucose uptake in skeletal muscle by increasing
blood flow to these tissues through a NO-dependent pathway
(33). In this scenario, allopurinol or benzbromarone may be
acting to prevent metabolic syndrome by blocking hyperuricemia-induced endothelial dysfunction.
Unlike glucose, the oral ingestion of fructose results in a
rapid increase in serum uric acid within 30 60 min in humans
(38). The mechanism by which fructose raises serum uric acid
has been previously studied. Fructose enters hepatocytes,
where it is rapidly phosphorylated by fructokinase to fructose1-phosphate (13). During this reaction, ATP donates the phos-
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