AJH 2000;13:547–551
Blood Pressure and Metabolic Changes
During Dietary l-Arginine Supplementation
in Humans
Alfonso Siani, Ermenegilda Pagano, Roberto Iacone, Licia Iacoviello, Francesco Scopacasa, and
Pasquale Strazzullo
Dietary L-arginine supplementation has been
proposed to reverse endothelial dysfunction in
such diverse pathophysiologic conditions as
hypercholesterolemia, coronary heart disease, and
some forms of animal hypertension. In particular,
chronic oral administration of L-arginine prevented
the blood pressure rise induced by sodium
chloride loading in salt-sensitive rats. To
investigate the effects of L-arginine–rich diets on
blood pressure and metabolic and coagulation
parameters we performed a single-blind,
controlled, crossover dietary intervention in six
healthy volunteers. The subjects (aged 39 ⴞ 4
years, body mass index [BMI] 26 ⴞ 1 kg/m2,
mean ⴞ SEM) received, in random sequence, three
different isocaloric diets, each for a period of 1
week (Diet 1: control; Diet 2: L-arginine enriched
by natural foods; Diet 3: identical to Diet 1 plus
oral L-arginine supplement). Sodium intake was set
at a constant level (about 180 mmol/day)
throughout the three study periods. A blood
Received April 16, 1999. Accepted September 22, 1999.
From the Epidemiology and Prevention Unit, Institute of Food
Sciences and Technology, National Research Council, Avellino; De-
partments of Clinical and Experimental Medicine, and Laboratory
Medicine, University of Naples “Federico II”, Naples; and Labora-
tory of Thrombosis Pharmacology, Consorzio Mario Negri Sud, S.
Maria, Imbaro, Italy.
© 2000 by the American Journal of Hypertension, Ltd.
Published by Elsevier Science, Inc.
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pressure decrease was observed with both L-
arginine-rich diets (Diet 2 v 1, SBP: ⴚ6.2 mm Hg
[95% CI: ⴚ0.5 to ⴚ11.8], DBP: ⴚ5.0 mm Hg [ⴚ2.8 to
ⴚ7.2]; Diet 3 v 1, SBP: ⴚ6.2 mm Hg [ⴚ1.8 to ⴚ10.5],
DBP: ⴚ6.8 mm Hg [ⴚ3.0 to ⴚ10.6]). A slight
increase in creatinine clearance (P ⴝ .07) and a fall
in fasting blood glucose (P ⴝ .008) occurred after
Diet 3 and, to a lesser extent, after Diet 2. Serum
total cholesterol (P ⴝ .06) and triglyceride (P ⴝ
.009) decreased and HDL cholesterol increased
(P ⴝ .04) after Diet 2, but not after Diet 3.
These results indicate that a moderate increase in
L-arginine significantly lowered blood pressure and
affected renal function and carbohydrate
metabolism in healthy volunteers. Am J
Hypertens 2000;13:547–551 © 2000 American
Journal of Hypertension, Ltd.
KEY WORDS: L-arginine, diet, blood pressure,
metabolism.
Bio-Arginina was a gift from Farmaceutici Damor s.p.a, Naples,
Italy. At the time of the study, Ermenegilda Pagano was recipient of
a National Research Council grant at the Institute of Food Science
and Technology.
Address correspondence and reprint requests to Alfonso Siani,
MD, Institute of Food Sciences and Technology, National Research
Council, Via Roma 52-83100, Avellino, Italy; e-mail: asiani@isa.av.cnr.it
0895-7061/00/$20.00
PII S0895-7061(99)00233-2
548 SIANI ET AL
A
lthough a few studies have investigated
the relationship between dietary protein
intake and blood pressure,1 little interest
has been paid thus far to the effect of
single amino acids. Recently, the identification of ni-
tric oxide (NO) as a product of the metabolism of
l-arginine through the NO synthase pathway2 has
opened a new avenue of research. l-arginine, a so-
called “conditionally” essential amino acid largely
present in several food items, plays a central role in a
number of major metabolic pathways (3). Its average
dietary intake is 5.4 grams/day, assuming a total daily
protein intake of 100 grams/day (3). On clinical
grounds, interest in the role of this aminoacid has been
stimulated by the observation that l-arginine supple-
mentation was able to attenuate the endothelial dys-
function associated with hypercholesterolemia (4) and
coronary heart disease (5). Moreover, hypertension
induced by sodium chloride loading in Dahl salt-sen-
sitive rats was prevented by chronic dietary l-arginine
supplementation.6 – 8 Yet in other studies the paren-
teral administration of large doses of l-arginine
acutely lowered blood pressure in normotensive9,11
and in salt-sensitive hypertensive individuals.9 –11
Against this background, the present study was de-
signed to investigate the effects of increased l-arginine
dietary intake—in the form of naturally arginine-rich
foods or as a pharmacologic preparation— on blood
pressure and metabolic and coagulation parameters in
healthy humans.
MATERIALS AND METHODS
Six untreated healthy volunteers (aged 39 ⫾ 4 years,
body mass index [BMI] 26 ⫾ 1 kg/m2, mean ⫾ SEM)
gave their informed consent to the study and, after
baseline evaluation, received, in random sequence,
three different isocaloric diets each for a period of 1
week.
Dietary Intervention The diets were tailored for
each patient by an expert dietitian to keep the caloric
intake constant . Diet 1 (control) was a relatively low
l-arginine diet (3.5– 4.0 g/day). Diet 2 was an l-argi-
nine– enriched diet (10 g/day) based on natural foods;
dry legumes (lentils) and nuts (hazelnuts, walnuts,
and peanuts) were chosen as major sources of l-argi-
nine, due to their particularly high l-arginine content.
Diet 3 was identical to Diet 1 (control diet), but was
supplemented with 10 g/day of an oral l-arginine
preparation (Bio-Arginina, Farmaceutici Damor s.p.a,
Naples, Italy), given three times a day. The nutrient
composition of the diets was calculated based upon
updated tables of food composition.12 The calculated
composition of the diets is reported in Table 1. The
three diets were similar with regard to their macronu-
trient composition, with the exception of fiber intake,
AJH–MAY 2000 –VOL. 13, NO. 5, PART 1
TABLE 1. CALCULATED COMPOSITIONS OF THE
CONTROL DIET AND THE INTERVENTION DIET
Diet 2
Diet 1 (L-Arginine
Nutrient (Control) Rich)
Total carbohydrate (% energy) 53 49
Protein (% energy) 19 22
Total fat (% energy) 28 29
Saturated (% energy) 8 6
Polyunsaturated (% energy) 3 3
Fiber (g/day) 22 47
l-Arginine (mg/day) 4080 9650
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which was higher in Diet 2. The dietary content of
l-arginine in Diet 2 was more than doubled as com-
pared with the control diet. Sodium intake was set at
a constant level throughout the three study periods
(about 180 mmol/day). Meals were prepared in the
research kitchen of the Institute of Food Sciences and
Technology in Avellino. On weekdays, the partici-
pants had lunch on site and took their evening meals
with them. On Friday, they received their weekend
meals to be consumed off site. Two participants
started the study with the control diet (Diet 1), two
with the Diet 2, and two with the Diet 3.
Measurements Blood pressure and anthropometric,
metabolic, and coagulation parameters were mea-
sured at baseline and at the end of each study period.
Blood pressure was measured by a trained observer
blind to the participant’s dietary regimen using a ran-
dom-zero sphygmomanometer (Gelman Hawksley
Ltd., Lancing, UK); after the participant had been sit-
ting upright for at least 10 min, systolic and fifth-phase
diastolic pressure were taken three times, 2 min apart.
The average of the last two measurements was used in
the analysis.
Serum cholesterol, triglycerides, and glucose were
measured with automated methods (Cobas-Mira,
Roche, Italy). High-density lipoprotein (HDL) choles-
terol was measured by the precipitation method. Cre-
atinine concentrations in blood and urine samples
were measured by the picric acid colorimetric method.
Urinary electrolytes were measured using an EA-2
Beckman Electrolyte Analyzer. Plasma insulin concen-
tration was measured by radioimmunoassay (Insulina
Lisophase, Technogenetics, Milano, Italy). Plasma fi-
brinolytic activity was measured on plasma euglobu-
lin fraction by the fibrin plate method (Sigma Chem-
icals Co., St. Louis, MO) and was expressed as the
euglobulin lysis area (ELA, mm2). PAI-1 antigen levels
and t-PA antigen levels were determined by commer-
cial double-antibody assay (American Diagnostica
Inc., Greenwich, CT). Fibrinogen was determined by a
AJH–MAY 2000 –VOL. 13, NO. 5, PART 1 DIETARY L-ARGININE AND BLOOD PRESSURE 549

TABLE 2. EFFECTS OF L-ARGININE–RICH DIETS ON BLOOD PRESSURE AND METABOLIC PARAMETERS

Diet 2 v Diet 1 Diet 3 v Diet 1,

Diet 1 Difference, M Difference, M
M (SEM) (95% CI) P (95% CI) P

SBP (mm Hg) 133.2 (2.7) ⫺6.2 (⫺0.5 to ⫺11.8) .03 ⫺6.2 (⫺1.8 to ⫺10.5) .01
DBP (mm Hg) 81.2 (4.6) ⫺5.0 (⫺2.8 to ⫺7.2) .002 ⫺6.8 (⫺3.0 to ⫺10.6) .006
Crea Cl (mL/s) 2.55 (0.31) 0.26 (⫺0.20 to 0.71) .20 0.30 (⫺0.03 to 0.63) .07
U-Na (mmol/24 h) 190 (18) ⫺8 (⫺57 to 41) .69 ⫺12 (⫺88 to 65) .71
U-K (mmol/24 h) 53 (7) 22 (⫺2 to 48) .06 1 (⫺7 to 8) .73
S-chol (mmol/L) 4.86 (0.36) ⫺0.33 (⫺0.67 to 0.001) .06 0.07 (⫺0.15 to 0.31) .45
S-TG (mmol/L) 0.93 (0.12) ⫺0.29 (⫺0.47 to ⫺0.11) .009 ⫺0.21 (⫺0.48 to 0.03) .08
HDL-Chol (mmol/L) 1.21 (0.11) 0.1 (0.005 to 0.20) .04 0.04 (⫺0.05 to 0.13) .32

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Plasma insulin (mU/L) 7.8 (0.7) 0.3 (⫺1.5 to 2.1) .68 0.4 (⫺0.9 to 1.6) .47
FBG (mmo/L) 4.81 (0.22) ⫺0.20 (⫺0.46 to 0.06) .10 ⫺0.44 (⫺0.71 to ⫺0.17) .008
SBP, systolic blood pressure; DBP, diastolic blood pressure; Crea Cl, creatinine clearance; U-Na, 24-h urinary Na excretion; U-K, 24-h urinary K
excretion; S-chol, serum total cholesterol; S-TG, serum triglyceride; HDL-chol, high-density lipoprotein cholesterol; FBG, fasting blood glucose; M, mean;
SEM, standard error of the mean; CI, confidence interval.

one-stage clotting assay (Hemoliance, Cologno, Mon- increase in 24-h potassium excretion was observed
zese, Italy). The concentration of fibrinogen was de- after Diet 2. A trend toward an increase in creatinine
termined by comparison with a reference curve. clearance was also observed at the end of both Diets 2
Statistical analysis Statistical analysis was carried and 3, the difference approaching statistical signifi-
out using the Statistical Package for Social Sciences cance versus the control diet after Diet 3.
(SPSS Italia, Bologna, Italy). Results are expressed as Body weight, plasma insulin levels, and coagulation
mean ⫾ SEM or 95% confidence interval (CI; where factors (ELA, PAI-I, t-PA, fibrinogen) did not change
appropriate). Differences between the arginine-rich di- throughout the study.
ets and the control diet were analyzed for statistical
significance by paired t test. The distributions of se-
rum glucose and triglycerides, as well as of plasma
insulin, were normalized by log transformation and
log-transformed values were used in the analysis.
Two-sided P values less than .05 were considered
statistically significant unless otherwise indicated.
RESULTS
The main results are summarized in Table 2. All sub-
jects but one had baseline blood pressure values
within the normal range. A significant average blood
pressure decrease was observed after both Diets 2 and
3, compared with the control diet. Figure 1 shows
changes in individual systolic and diastolic blood
pressure values; the increase in l-arginine intake, both
from dietary sources and from pharmacologic supple-
ments, resulted in a blood pressure fall in all subjects.
The decrease in blood pressure was not significantly
related to the initial blood pressure value.
After Diet 3, a significant reduction was observed in
fasting blood glucose concentration; a similar trend
was also apparent after Diet 2. A decrease in serum
total cholesterol and triglycerides and an increase in
HDL-cholesterol concentration were observed after FIGURE 1. Individual systolic and diastolic blood pressure
Diet 2. Twenty-four– hour urinary sodium excretion changes after l-arginine supplemented diets (A: diet 2 vs diet 1; B:
was very close to the prefixed value (180 mmol/day) diet 3 vs diet 1). SBP: systolic blood pressure; DBP: diastolic blood
in all dietary periods, whereas a trend toward an pressure.
550 SIANI ET AL
DISCUSSION
To the best of our knowledge, there are no published
data from controlled studies dealing with the blood
pressure effects of l-arginine–rich diets in humans.
The main finding of our work is that this dietary
intervention was associated with a statistically and
biologically significant blood pressure reduction,
whether l-arginine was provided through natural
foods or as a pharmacologic preparation.
A reduction in serum glucose and an apparent in-
crease in renal glomerular filtration rate (as suggested
by the higher creatinine clearance) were also observed.
To our knowledge, this is also the first report of an
effect of dietary l-arginine supplementation on glu-
cose metabolism in humans. An effect of l-arginine on
insulin release has been observed in acute experiments
using the parenteral route13; it is possible that a similar
mechanism accounts for our finding during chronic
oral administration, although no significant changes in
fasting plasma insulin levels were observed in the
present study. The increase in creatinine clearance is
consistent with the recognized effects of an oral pro-
tein load or an infusion of amino acid mixtures on
renal hemodynamics, although here again the avail-
able information stems essentially from acute experi-
ments.14,15 As to the improvement in serum lipid pro-
file after Diet 2, this was most likely due to the higher
fiber content of that diet.16
The explanation for the blood-pressure–lowering ef-
fect of l-arginine administration in our study is not
apparent. Modulation of NO production may be crit-
ical for blood pressure control, particularly during
dietary salt loading,11,17 and l-arginine feeding may
actually correct a failure in NO production under
these circumstances.6 – 8,10,11 A dysfunction in the l-
arginine-nitric oxide pathway in the renal circulation
has also been advocated by recent studies in patients
with mild primary hypertension.9 –11 Interestingly, in
two of these studies9,11 the acute infusion of l-arginine
reduced blood pressure also in normotensive control
subjects, a finding consistent with our results. An un-
resolved issue, however, is that increased dietary l-
arginine availability does not necessarily lead to en-
hanced NO production, as l-arginine supply is not
normally rate limiting for NO synthesis.18 Alternative
or additional mechanisms whereby l-arginine may
affect blood pressure and renal function need to be
investigated (direct effects of the amino acid, release of
hormones, prostaglandins, etc.).
A few potential limitations of this study should be
considered. First, the short-term duration of the inter-
vention does not allow exclusion of the possibility that
the blood-pressure–lowering effect of l-arginine may
not be sustained over time. Second, we set out a high
level of sodium intake for our experimental diets on
AJH–MAY 2000 –VOL. 13, NO. 5, PART 1
the assumption that modulation of NO delivery is
critical to blood pressure control during dietary salt
loading.10,11,17 Thus we cannot exclude that the re-
sponse may be limited to (or greater in) subjects eating
a high-sodium diet, as we did not study subjects at
different levels of sodium intake. Third, the possibility
that dietary components other than l-arginine might
at least partly explain the effect observed needs to be
considered as well. The 24-h urinary potassium excre-
tion of our participants increased after Diet 2, most
likely due to the increase in legume and nut intake. In
a previous long-term controlled trial we showed that
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increasing dietary potassium intake improved blood
pressure control in hypertensive patients19; however,
in the present study, a blood pressure fall similar to
the one observed with Diet 2 took place when l-
arginine was given as pharmacologic supplementa-
tion to the control diet, thus ruling out the confound-
ing influence of dietary components other than
l-arginine. Finally, the dietary modifications adopted
in our trial are not intended for use in clinical practice.
Our dietary modifications were implemented to
achieve the goal of consistently increasing l-arginine
intake while avoiding, as much as possible, the influ-
ence of confounding factors on the experimental re-
sults. Interestingly, however, the magnitude of the
blood pressure fall observed with both dietary and
pharmacologic increase in l-arginine intake was sim-
ilar to that reported by the DASH Collaborative Re-
search Group20 and by McCarron et al,21 as a conse-
quence of more complex dietary modifications aimed
to provide, respectively, a dietary model for the pre-
vention and treatment of high blood pressure 20 and
for the nutritional management of cardiovascular risk
factors.21
In conclusion, the present study indicates that an
approximately twofold increase in dietary l-arginine
intake had significant hemodynamic and metabolic
effects in a group of healthy men. Further studies are
warranted to confirm these findings over a long-term
period and in a larger population; to evaluate the
blood-pressure–lowering effect of oral l-arginine in
hypertensive patients; and to elucidate the mecha-
nism(s) involved.
ACKNOWLEDGMENTS
We thank Rosalba Giacco, MD, and Ms. Anna Maria
Palumbo for the generous help with the fieldwork; Angela
Giacco, RD, and Delia Pacioni, RD, for valuable dietary
advice; and Rosanna Scala for editing the manuscript.
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