Effects of unripe grape juice (verjuice) on plasma lipid profile, blood pressure, malondialdehyde and total antioxidant capacity in normal, hyperlipidemic and hyperlipidemic with hypertensive human volunteers

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Journal of Medicinal Plants Research Vol. 6(45), pp. 5677-5683, 25 November, 2012
Available online at http://www.academicjournals.org/JMPR
DOI: 10.5897/JMPR11-1146 ISSN 1996-0875 ©2012 Academic Journals
Full Length Research Paper
Effects of unripe grape juice (verjuice) on plasma lipid
profile, blood pressure, malondialdehyde and total
antioxidant capacity in normal, hyperlipidemic and
hyperlipidemic with hypertensive human volunteers
Mohsen Alipour1, Parvin Davoudi2 and Zahra Davoudi3*
1Department of Physiology and Pharmacology, Faculty of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran.
2Department of Biology, Payam Noor University, Zanjan, Iran.
3Department of Biotechnology, Faculty of Advanced Biomedical Sciences, Tabriz University of Medical Sciences,
Tabriz Iran.
Accepted 1 November, 2011
There is a widespread belief in Iranian society that verjuice consumption has useful effects on blood
lipid profile and hypertension. This study was designed to examine this hypothesis. This study subjects
included three groups: (A) healthy individuals volunteers of age 20 to 30 years; (B) hyperlipidemic
patients of age 30 to 60 years old; and (C) hyperlipidemic plus hypertensive patients of age 30 to 60
years. All subjects were asked to consume 200 ml of verjuice twice per day for one month. At the
beginning of this study and then every two weeks, blood pressure and heart rate were measured. Blood
sample were also collected and all parameters consisted of plasma lipid profile, total antioxidant
capacity (TAC), and malondialdehyde (MDA) were measured. There was no significant difference
between the levels of lipid profile components, heart rate, and blood pressure before and after
consumption of the verjuice in healthy volunteers. However, in all the three groups, the concentration
of TAC and high density lipoprotein cholesterol (HDL-C) to low density lipoprotein cholesterol (LDL-C)
ratio was increased and MDA concentration was reduced respectively, in all subjects after verjuice
consumption. In hyperlipidemic and hyperlipidemic plus hypertensive peoples, administration of
verjuice, after four weeks (no two weeks) resulted in significant reduction of blood pressure along with
the reduction of LDL-C, triglyceride (TG), and total cholesterol (TC) concentrations. In conclusion,
although blood pressure and lipid profile disorders did not completely return to normal levels by
verjuice consumption, it seems that it is used as a flavoring, not devoid of benefit may be due to its
antioxidant effects.
Key words: Verjuice, blood pressure, blood lipid profile, malondialdehyde (MDA), total antioxidant capacity
(TAC).
INTRODUCTION
Hypertension (HTN) is a progressive cardiovascular (CV)
syndrome arising from complex and interrelated
etiologies (Giles et al., 2011). It is the most common form
of CV disease, affecting millions of people throughout the
*Corresponding author. E-mail: zahrahdavoudi67@yahoo.com.
Tel: +98 241 4240301-3. Fax: +98 2414249553.
world and about 20% of the adult population in many
countries. HTN is interconnected with coronary artery
disease, stroke, congestive heart failure, and renal
dysfunction. HTN, as one of the major risk factors for CV-
related mortality accounts for 20 to 50% of all deaths
(Yang et al., 2004). HTN is defined as a constantly
elevated blood pressure further than 140 over 90 mmHg,
a systolic pressure above 140 with a diastolic pressure
above 90 mmHg. Chronic HTN is a "silent" condition; it

5678 J. Med. Plants Res.
has no symptoms and is the leading cause of many life
threatening or disabling diseases. Many people have
HTN without knowing it. Therefore, it is important to have
blood pressure checked regularly. Blood pressure
measurements are classified in stages, according to
severity. Normal blood pressure: less than 120/80 mmHg,
pre-HTN: 120 to 129/80 to 89 mmHg, stage 1 HTN: 140
to 159/90 to 99 mmHg and stage 2 HTN: at/or greater
than 160 to 179/100 to 109 mmHg (Giles et al., 2011).
Although, many factors including age, sex, race, heredity,
self sensitivity, obesity, inactive life style, etc., are
involved in HTN (secondary HTN); nevertheless, the
cause of most HTN is not known (Primary or essential
HTN) (Reaven et al., 1996). Adequate treatment of HTN
can diminish health risks, recover quality and quantity of
life, and reduce overall costs of health care. Treatment to
lower blood pressure may include dropping salt intake,
reducing fat intake, losing weight, getting regular
exercise, quitting smoking, reducing alcohol con-
sumption, managing stress, and taking antihypertensive
medications (Sacks et al., 2001).
For many years, it has been a widespread belief in our
society that certain foods and herbal drugs have useful
effects for lipid and blood pressure lowering and several
studies also established the efficacy of some of them
(Aminian et al., 2006; Caron and White, 2001). Unripe
grape juice (verjuice or verjus) is one of these agents.
Verjuice is an unfermented green grape juice obtained by
directly pressing green grapes. It has a unique flavor and
sour taste. Verjuice is used as an alternative to vinegar
and lemon juice (Aminian et al., 2003). Verjuice is
produced and consumed locally, particularly in the
Mediterranean, Southeastern regions of Turkey, and
many different regions of Iran. It has been used to
augment the flavor of traditional meals, salads, and
appetizers, and as an ingredient in the production of
various drinks and several sausages such as mustard
sausage (Hildebrandt and Matchuk, 2002; Hayoglui et al.,
2009). The use of verjuice as a food ingredient and as a
medicine has a long history, from 370 to 460 B.C until
today (Pour Nikfardjam, 2008). Verjuice, which is known
as "Abe ghureh" in Iran, is still widely used for cooking in
our country. According to Iranian folk medicine, verjuice
has antilipidemic and antihypertensive effects. The
interest of Western countries has been attracted to this
product in recent years. Although, some information are
currently available on the chemical composition and
polyphenolic composition of this food stuff; however, few
studies have been conducted regarding the medical use
of verjuice, which analyze the effect of its ingestion on
health. Aminian et al. (2006) have investigated the impact
of verjuice on plasma lipid level in rabbit. Their results
show that verjuice has no preventive or therapeutic effect
in hypercholesterolemia (Pour Nikfardjam, 2008). To the
best of our knowledge, very limited information is avail-
able regarding the effect of verjuice on blood pressure,
plasma lipid profile, and oxidative stress indices in
humans. Thus, this work aimed to investigate the effects
of 4 weeks Iranian verjuice ingestion on blood pressure,
plasma lipid profile, total antioxidant capacity (TAC) and
malondialdehyde (MDA) in normal, hyperlipidemic, and
hypertensive human volunteers.
SUBJECTS AND METHODS
This study was carried out in the Department of Physiology, Zanjan
University of Medical Sciences and Department of Biology, Zanjan
Payam Noor University, Zanjan, Iran. The study of verjuice effects
on HTN, lipid profile TAC, and MDA was conducted on three groups
of people. Group 1: normal volunteers, thirteen male of healthy and
nonsmoking volunteers (Students of Payam Noor University) aged
between 20 and 30 years, who were under no medication and
without any history of disease. Group 2: this group included 11
male patients of age 30 to 60 years old, who had abnormal blood
lipids and normal blood pressure, but they were unaware of their
blood lipid disorders. Group 3: this group included 7 male patients
of age 30 to 60 years old, who had high blood pressure and
abnormal blood lipids as well. They were all aware of their
condition, previously. Adequate and accurate description was given
to all the people about their condition and the research. Written
consent was obtained from all the participants.
Verjuice preparation and consumption
Unripe green grapes were bought from the market and its juice was
extracted by means of pressing grapes using the juice making
machine and the juice obtained was sieved through a nonmetal
filter. The juice was transferred into bottles and was kept in
refrigerator for consumption. All subjects were asked to consume
200 ml of verjuice twice per day (night, before sleeping and
morning, after waking up) for one month. All persons sustained their
habitual diets throughout the study. At three time intervals, the start
of the study and after two and four weeks of the verjuice
consumption, blood pressure and heart rate was measured. Also,
blood samples (15 ml) were collected in the morning from each
individual after 12 h of fasting and plasma was immediately
separated by centrifugation at 5000 rpm for 15 min at room
temperature for determination of biochemical parameters.
Blood pressure measurement and heart rate
Baseline, blood pressure was measured by the auscultatory
method with a random zero mercury sphygmomanometer and
standard cuff (12 × 34 cm). The blood pressure measurement was
taken in the seated position, quietly in a chair with feet on the floor
and an arm support at the heart level. After recording the systolic
and diastolic pressures, mean arterial pressures (MAP) was
calculated by the following formula: MAP = diastolic pressure +
(systolic pressure - diastolic pressure) / 3 (Bern and Levy, 2010).
Heart rate (HR) was also measured by finding the pulse of the
radial artery via pressuring the thumb on the ventral aspect of the
wrist (Sharma et al., 2011).
Biochemical measurements
Blood samples were drawn after fasting for 12 h before verjuice
administration in order to plasma lipid profile, MDA, and TAC
measurements. All persons continued their habitual diets during the
study.

Alipour et al. 5679
Table 1. Plasma lipid levels (mg/dl), MAP (mmHg), and HR in human healthy volunteers during one month verjuice
consumption.
Variable
Start (Mean ± SD)
2 weeks (Mean ± SD)
4 weeks (Mean ± SD)
p-value
LDL-C
170.9 ± 7.2
168.4 ± 7.3
165.5 ± 7.4
0.04
HDL-C
48.2 ± 4.9
49.6 ± 4.9
50.7 ± 5
0.45
HDL-C/LDL-C
0.28 ± 0.03
0.29 ± 0.03
0.30 ± 0.03
0.13
TG
180.6 ± 16
178.2 ± 15.2
175.3 ± 14.7
0.65
TC
254.7 ± 3.8
252.3 ± 4.1
249.5 ± 4.5
0.04
MAP
94.7 ± 2.2
94.6 ± 1.6
94.1 ± 1.6
0.56
RH
74.8 ± 1.9
74.2 ± 1.9
73.8 ± 1.8
0.41
Measurements were performed at the start (before verjuice consumption) and every 2 weeks (after start of verjuice
consumption). TC, Total cholesterol; LDL-C, low-density lipoprotein cholesterol; HDL-C, high-density lipoprotein
cholesterol; TG, triglyceride; MAP, mean arterial pressure; HR, heart rate. Significant level (p < 0.05).
Plasma lipid profile including total cholesterol (TC), low density
lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol
(HDL-C) and triglyceride (TG) were determined by enzymatic
methods using automatic analyzer (Abbott, Alcyon 300, USA).
Plasma TAC concentration was determined in 600 nm at 37°C
against air by spectrophotometer, according to the guidelines
coming with the relevant kit (TAC kit, Randox Co, Germany) and its
concentration was calculated by the related formula (kit direction)
and was expressed as mmol/L. The amount of malondialdehyde
(MDA) was determined by thiobarbituric acid (TBA) assay. All
reagents that were used in this assay were obtained from Merck
(Darmstadt, Germany). Briefly, 0.50 ml of plasma was added to 3
ml of 1% phosphoric acid, 1 ml of 0.60% TBA, and 0.15 ml of
0.20% butylated hydroxytoluene in 95% methanol. The samples
were heated in a boiling water bath for 45 min, cooled, and 4 ml of
1-butanol was added. The butanol phase was separated by
centrifugation at 3000 rpm for 10 min, and absorbance was
measured at 532 nm. The concentration of MDA was calculated
and was expressed as μM (Alipour et al., 2006).
Statistical analyses
Data are expressed as the mean ± SD. All statistical analyses were
performed using Kruskal–Wallis test, followed by the Mann–
Whitney U test (Statistical Package for Social Sciences (SPSS)
software version 16). In all statistical evaluations, p < 0.05 was
considered as the criterion for statistical significance.
RESULTS
Based on our results, the plasma levels of LDL-C, HDL-
C, TG, and TC after one month verjuice consumption
(200 ml twice per day) were not significantly lesser than
the equivalent levels before the verjuice consumption in
healthy volunteers. Also, no significant difference was
observed between the blood pressure and heart rate
before and after verjuice consumption (Table 1).
However, in these subjects, the levels of TAC for two and
four weeks after starting the trial (verjuice consumption),
were significantly higher than the corresponding levels
before the verjuice consumption (p = 0.005). In contrast
to TAC and MDA concentration, an index of lipid
peroxidation was significantly reduced (p = 0.005) in
response to 4 weeks verjuice consumption (Figure 1).
Table 2 shows the plasma LDL-C, HDL-C, TG, and TC
concentrations in hyperlipidemic subjects (group II)
before and two and four weeks after verjuice consumpt-
ion. Means comparison showed significant differences
only in TC and LDL fractions. Significant reduction was
observed in plasma levels of LDL-C (p = 0.019) and TC
(p = 0.016) in hyperlipidemic subjects receiving verjuice
for four weeks. Positive changes but non-significant
statistically was found in other parameters including HDL-
C, TG, and HDL-C/LDL-C. MDA values were high in
hyperlipidemic subjects when compared with the normal
subjects (Figure 2). Mean amounts of MDA was reduced
noticeably (p = 0.005) after 2 and 4 weeks of verjuice
consumption in hyperlipidemic subjects than before
verjuice consumption (Figure 2). Unlike MDA, a signify-
cant increase was found in plasma levels of TAC after 4
weeks of verjuice consumption. Reduction of MDA and
elevation of TAC levels were depended on the duration of
the verjuice consumption. That is, MDA concentration
was lower after 4 weeks of verjuice consumption than 2
weeks. The same results, but unlike the MDA, was seen
for TAC levels (Figure 2).
In group 3 (hyperlipidemic along with hypertension
subjects), in addition to the significant decrease of LDL-C
and TC, that were also seen in the previous trial (group 2;
HDL-C concentration and the proportion of HDL-C to
LDL-C were significantly increased following the
consumption of verjuice as compared to starting stage (p
= 0.002 to 0.007)) (Table 3). A significant reduction and a
significant elevation were found in plasma levels of MDA
and TAC, respectively, in subjects receiving verjuice for 2
and 4 weeks (Figure 3).
Interestingly, 2 and 4 weeks of verjuice consumption
resulted in the significant reduction of mean arterial
pressure in hypertensive-hyperlipidemic subjects in
comparison to the starting stage (before verjuice
consumption) (p = 0.003, p = 0.005, respectively). There
was no statistically significant difference between the
heart rate at the start of the study and following 2 and 4
weeks of verjuice consumption.

5680 J. Med. Plants Res.
TAC
wks4wks2Start
Concentration (micromol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
wks4wks2Start
Concentration (mmol/L)
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
wks4wks2Start
Concentration (micromol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
wks4wks2Start
Concentration (mmol/L)
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
wks4wks2Start
Concentration (micromol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
wks4wks2Start
Concentration (mmol/L)
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
MDA
wks4wks2Start
Concentration (micromol/L)
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
wks4wks2Start
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
wks4wks2Start
Concentration (micromol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
wks4wks2Start
Concentration (mmol/L)
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
MDA
wks4wks2Start
Concentration (micromol/L)
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
wks4wks2Start
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
Figure 1. TAC (left) and MDA (right) concentrations before and after 2 and 4 weeks of verjuice consumption in healthy volunteers.
The concentration of TAC increased during verjuice consumption as compared to before consumption (p < 0.005). The reduction of
MDA concentration was significant (p < 0.005) after 4 weeks of verjuice consumption. Data are mean ± SD. Abbreviations: TAC,
total antioxidant capacity; MDA, malondialdehyde; wks, weeks.
Table 2. Comparison of plasma lipid levels (mg/dl), MAP (mmHg) and HR in hyperlipidemic subjects before and
after verjuice consumption (every 2 weeks).
Variable
Start (Mean ± SD)
2 weeks (Mean ± SD)
4 weeks (Mean ± SD)
p-value
LDL-C
170.9 ± 7.2
168.4 ± 7.3
165.5 ± 7.4
0.04
HDL-C
48.2 ± 4.9
49.6 ± 4.9
50.7 ± 5
0.45
HDL-C/LDL-C
0.28 ± 0.03
0.29 ± 0.03
0.30 ± 0.03
0.13
TG
180.6 ± 16
178.2 ± 15.2
175.3 ± 14.7
0.65
TC
254.7 ± 3.8
252.3 ± 4.1
249.5 ± 4.5
0.04
MAP
94.7 ± 2.2
94.6 ± 1.6
94.1 ± 1.6
0.56
RH
74.8 ± 1.9
74.2 ± 1.9
73.8 ± 1.8
0.41
Measurements were performed at the start (before verjuice consumption) and every 2 weeks (after start of verjuice
consumption). TC, Total cholesterol; LDL-C, low-density lipoprotein cholesterol; HDL-C, high-density lipoprotein
cholesterol; TG, triglyceride; MAP, mean arterial pressure; HR, heart rate; wks, weeks. Significant level (p < 0.05)
Table 3. Comparison of plasma lipid levels (mg/dl), MAP (mmHg) and HR in hyperlipidemic plus subjects before and
after verjuice consumption (every 2 weeks).
Variable
Start (Mean ± SD)
2 weeks (Mean ± SD)
4 weeks (Mean ± SD)
p-value
LDL-C
210 ± 13.5
201.6 ± 10.1
194.7 ± 7.4
0.05
HDL-C
51.6 ± 2.6
57.6 ± 1.9
59.9 ± 2.2
0.001
HDL-C/LDL-C
0.25 ± 0.02
0.28 ± 0.02
0.31 ± 0.01
0.001
TG
212.9 ± 14
208.7 ± 13.9
203.6 ± 12.9
0.35
TC
268.4 ± 44.4
246.3 ± 13.4
240.2 ± 13.5
0.001
MAP
111.1 ± 2.1
107.1 ± 1.8
105.9 ± 2
0.002
RH
84.1 ± 6.1
83.3 ± 5.9
81.6 ± 5.1
0.64
Measurements were performed at the start (before verjuice consumption) and every 2 weeks (after start of verjuice
consumption). TC, Total cholesterol; LDL-C, low-density lipoprotein cholesterol; HDL-C, high-density lipoprotein cholesterol;
TG, triglyceride; MAP, mean arterial pressure; HR, heart rate. Significant level (p < 0.05)

Alipour et al. 5681
TAC
wks4wks2Start
Concentration (micromol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
wks4wks2Start
Concentration (mmol/L)
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
MDA
wks4wks2Start
Concentration (micromol/L)
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
wks4wks2Start
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
wks4wks2Start
Concentration (micromol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
wks4wks2Start
Concentration (mmol/L)
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
MDA
wks4wks2Start
Concentration (micromol/L)
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
wks4wks2Start
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
wks4wks2Start
Concentration (micromol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
wks4wks2Start
Concentration (mmol/L)
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
MDA
wks4wks2Start
Concentration (micromol/L)
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
wks4wks2Start
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
wks4wks2Start
Concentration (micromol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
wks4wks2Start
Concentration (mmol/L)
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
MDA
wks4wks2Start
Concentration (micromol/L)
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
wks4wks2Start
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
Figure 2. TAC (left) and MDA (right) concentrations before and after 2 and 4 weeks of verjuice consumption in
hyperlipidemic subjects. Concentration of TAC was augmented in response to 4 weeks of verjuice consumption compared
to start stage (p = 0.005). A significant reduction in MDA concentration was seen after 2 and 4 weeks of verjuice
consumption compared with the start stage (p = 0.005). Data are mean ± SD. TAC, total antioxidant capacity; MDA,
malondialdehyde; wks, weeks.
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
wks4wks2Start
Concentration (micromol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
wks4wks2Start
Concentration (mmol/L)
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
MDA
wks4wks2Start
Concentration (micromol/L)
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
wks4wks2Start
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
wks4wks2Start
Concentration (micromol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
wks4wks2Start
Concentration (mmol/L)
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
MDA
wks4wks2Start
Concentration (micromol/L)
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
wks4wks2Start
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
TAC
4Wks2WksStart
Concentration (mmol/L)
2.2
2.0
1.8
1.6
1.4
1.2
1.0
M + 1 SE
M - 1 SE
Mean
MDA
4Wks2WksStart
Concentration (micromol/L)
3.5
3.0
2.5
2.0
1.5
1.0
.5
0.0
M + 1 SE
M - 1 SE
Mean
Figure 3. TAC (left) and MDA (right) concentrations before and after 2 and 4 weeks of verjuice consumption in
hyperlipidemic along with hypertension subjects. Concentration of TAC was augmented in response to 2 and 4 weeks of
verjuice consumption compared to start stage (p = 0.005). A significant reduction in MDA concentration was seen after 2 and
4 weeks of verjuice consumption compared with the start stage (p = 0.005). Data are mean ± SD. TAC, total antioxidant
capacity; MDA, malondialdehyde; wks, weeks.
DISCUSSION
The major findings of this study were that consumption of
verjuice for one month ameliorates mean arterial pressure
that was with the reduction of MDA as an indirect
measure of oxidative stress and elevation of TAC. In
addition, positive changes were observed in plasma lipid
profile including decrease of LDL-C and TC and increase

5682 J. Med. Plants Res.
of HDL-C and HDL-C to LDL-C ratio.
The consumption of verjuice as a food flavoring and as
a medicine has a long history. It has been used as food
ingredient and as a medicine for treatment of ulcers and
as a digestive agent after the ingestion of fatty foodstuff
such as brain and especially a particular type of food
known as "kalah-pacheh" in Iran (Pour Nikfardjam, 2008).
Despite the widespread uses of verjuice, limited scientific
information regarding the chemical composition and
particularly about its effects on physiological and
pathological conditions such as hypertension and lipid
profile are available. Aminian et al. (2006) studied the
effect of verjuice on plasma lipid levels in rabbits after
egg yolk ingestion. In contrast to our findings with regard
to the effect of verjuice as a lipid lowering agent, their
results show that verjuice has no preventive or thera-
peutic effect in hypercholesterolemia. In human studies, it
has been reported that verjuice has no lipid-lowering
effect. Based on their work, in hyperlipidemic subjects,
who had received 80 ml verjuice daily at lunch time for
the 4 months, TC and TG decreased significantly at the
end of the study, but authors suggested that this result
may be the result from the effect of diet and not verjuice
(Aminian et al., 2003).
HDL-C is known to be a significant and independent
predictor of coronary heart disease (CHD) risk (Eccleston
et al., 2002). A significant increase was found in plasma
levels of HDL-C in subjects receiving verjuice for 4
weeks. However, Aminian et al. (2006) showed that
verjuice consumption had no incremental effect on serum
HDL-C levels in healthy individuals and hyperlipidemic
subjects. To the best of our knowledge, regardless limited
studies of Aminian et al. (2006), no animal or human
study regarding the effect of verjuice consumption on
plasma lipid profile have been published. On the other
hand, few studies show the effect of grape juice on
hyperlipidemia. In support of our results, it has been
reported that red grape juice consumption is associated
with parallel reduction in the LDL-C and elevation in HDL-
C concentrations (Castilla et al., 2006). However,
Coimbra et al. (2005) reported no effect on HDL-C levels
after consumption of purple grape juice in healthy
persons for 14 days. In addition, red grape juice consum-
ption is coupled with significantly lower cholesterol/HDL
as atherogenic index, which is usually used as the best
lipid parameter for determining human heart disease risk
(Vinson et al., 2001).
In vitro studies showed that grape juice has significant
antioxidant activity and can inhibit the oxidation of LDL,
and consequently, nonalcoholic red grape extract may
have a similar beneficial effect to red wine (Day et al.,
1997). According to a few published studies, general
composition and polyphenolic content of verjuice show
various differences in many regions and countries.
Polyphenolic content of Iranian verjuice reported about
780 to 1330 mg/L (Pour Nikfardjam, 2008). Discrepancies
between our results and other studies (related to lipid
profile) may be connected to differences in the
composition and polyphenols content of verjuice, time of
consumption, and amount of consumption. Grape juice
and verjuice are very similar in structure and both have a
flavonoid component, which may have an antioxidant
effect on plasma lipoproteins and other molecules. This
study was designed to assess whether verjuice
consumption could in particular contribute to the MDA
decrease and TAC increase as indicators of oxidative
stress conditions and changes in lipid profile. In this
study, we observed a significant increase in the MDA
level of hyperlipidemic and hypertensive patients before
treatment with verjuice which suggests the presence of
increased oxidant stress. The total antioxidant status in
these patients decreased significantly in comparison to
the after treatment with verjuice. Therefore, it seems that
a negative correlation between the total antioxidant status
and lipid peroxidation in the plasma of healthy,
hyperlipidemic, and hyperlipidemic-hypertensive patients.
MDA reflects both autoxidation and oxygen mediated
peroxidation of polyunsaturated fatty acids in particular. It
reflects the oxidative status of the biological system. MDA
causes damage to LDL which in turn forms foam cells
finally. Due to increased production of reactive oxygen
species (ROS) and increased oxidative stress, lipid
peroxidation products are found to be elevated in
hyperlipidemic and hypertensive patients (Meera, 2011).
In healthy conditions, a balance exists between free-
radical generation and antioxidant defense system which
prevents occurrence of disease. This study implies that
hyperlipidemia and hypertension shift the balance in favor
of free-radical generation which leads to oxidative tissue
damage (Das et al., 2000), whereas, it seems the
verjuice consumption increases the total antioxidant
capacity and eliminates the harmful effects of free
radicals. Thus, we can conclude that verjuice has
antioxidant properties, and its beneficial effects may be
related to this effect.
To the best of our knowledge, any information is
currently available about the effects of verjuice
consumption on plasma levels of MDA and TAC in
normal subjects, hyperlipidemia, and hypertension in both
animals and humans.
Furthermore, this is the first study that investigates the
effect of verjuice consumption on blood pressure in
human. Hypertension is linked with a number of alter-
ations in the vascular system, including augmentation of
vascular tone, amplified shear stress, and activation of
the sympathetic nervous and renin-angiotensin-
aldosterone systems, because these functional and
structural changes promote endothelial damage. Hyper-
lipidemia is prevalent in hypertension, but the cause of
this association is unknown.
Impaired endothelial function is associated with
increased total-C/HDL-C values, perhaps as the result of
increased vascular oxidative stress and inflammation
(Sugiura et al., 2011). Oxidative stress may contribute to

the production and/or preservation of hypertension via
the number of possible mechanisms, including quenching
of the vasodilator nitric oxide (NO) by ROS such as
superoxide, production of vasoconstrictor lipid
peroxidation products, such as F2-isoprostanes and
MDA, diminution of tetrahydrobiopterin (BH4), an
important NO synthase (NOS) cofactor, as well as
structural and functional alterations within the
vasculature. These vascular changes may be mediated in
different ways (Ward and Croft, 2006). In this study,
plasma TAC level was significantly reduced and plasma
MDA level was significantly raised in hyperlipidemic and
hyperlipidemic with hypertension. These changes
suggest an association between increased oxidative
stress and the mentioned disorders. Reverse relations in
MDA and TAC levels that were observed following
consumption of verjuice may be attributed to the
antioxidative properties of verjuice which resulted in the
reduction of oxidative stress and thus improvement of
blood pressure.
In conclusion, this study indicates that the consumption of
verjuice resulted in noticeable raise in TAC levels and
marked reduction in MDA levels in hyperlipidemic and
hyperlipidemic with hypertensive patients, which are
associated with the reduction of mean arterial pressure.
Although, blood pressure and lipid profile disorders did
not completely return to normal levels by verjuice
consumption; it seems that its drinking as a flavoring, is
not devoid of benefits, may be due to its antioxidant
properties.
ACKNOWLEDGMENT
The authors wish to express their gratitude to Dr. Mosavi
Nasb and Mr. Fallah for kind help in the statistical
analysis of data. This study was supported by grant from
the PyameNoor University of Zanjan, Zanjan, Iran.
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