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Apple Cider Vinegar Modulates Serum Lipid Profile, Erythrocyte, Kidney, and Liver Membrane Oxidative Stress in Ovariectomized Mice Fed High Cholesterol


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The purpose of this study was to investigate the potentially beneficial effects of apple cider vinegar (ACV) supplementation on serum triglycerides, total cholesterol, liver and kidney membrane lipid peroxidation, and antioxidant levels in ovariectomized (OVX) mice fed high cholesterol. Four groups of ten female mice were treated as follows: Group I received no treatment and was used as control. Group II was OVX mice. Group III received ACV intragastrically (0.6 % of feed), and group IV was OVX and was treated with ACV as described for group III. The treatment was continued for 28 days, during which the mice were fed a high-cholesterol diet. The lipid peroxidation levels in erythrocyte, liver and kidney, triglycerides, total, and VLDL cholesterol levels in serum were higher in the OVX group than in groups III and IV. The levels of vitamin E in liver, the kidney and erythrocyte glutathione peroxidase (GSH-Px), and erythrocyte-reduced glutathione (GSH) were decreased in group II. The GSH-Px, vitamin C, E, and β-carotene, and the erythrocyte GSH and GSH-Px values were higher in kidney of groups III and IV, but in liver the vitamin E and β-carotene concentrations were decreased. In conclusion, ACV induced a protective effect against erythrocyte, kidney, and liver oxidative injury, and lowered the serum lipid levels in mice fed high cholesterol, suggesting that it possesses oxidative stress scavenging effects, inhibits lipid peroxidation, and increases the levels of antioxidant enzymes and vitamin.
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Apple Cider Vinegar Modulates Serum Lipid Profile, Erythrocyte,
Kidney, and Liver Membrane Oxidative Stress in Ovariectomized
Mice Fed High Cholesterol
Mustafa Nazırog
˘lu Mustafa Gu
Cemil O
¨zalp Saydam
Mustafa Ku
¨kayaz Ercan So
Received: 8 March 2014 / Accepted: 13 May 2014 / Published online: 4 June 2014
ÓSpringer Science+Business Media New York 2014
Abstract The purpose of this study was to investigate the
potentially beneficial effects of apple cider vinegar (ACV)
supplementation on serum triglycerides, total cholesterol,
liver and kidney membrane lipid peroxidation, and anti-
oxidant levels in ovariectomized (OVX) mice fed high
cholesterol. Four groups of ten female mice were treated as
follows: Group I received no treatment and was used as
control. Group II was OVX mice. Group III received ACV
intragastrically (0.6 % of feed), and group IV was OVX
and was treated with ACV as described for group III. The
treatment was continued for 28 days, during which the
mice were fed a high-cholesterol diet. The lipid peroxida-
tion levels in erythrocyte, liver and kidney, triglycerides,
total, and VLDL cholesterol levels in serum were higher in
the OVX group than in groups III and IV. The levels of
vitamin E in liver, the kidney and erythrocyte glutathione
peroxidase (GSH-Px), and erythrocyte-reduced glutathione
(GSH) were decreased in group II. The GSH-Px, vitamin
C, E, and b-carotene, and the erythrocyte GSH and GSH-
Px values were higher in kidney of groups III and IV, but in
liver the vitamin E and b-carotene concentrations were
decreased. In conclusion, ACV induced a protective effect
against erythrocyte, kidney, and liver oxidative injury, and
lowered the serum lipid levels in mice fed high cholesterol,
suggesting that it possesses oxidative stress scavenging
effects, inhibits lipid peroxidation, and increases the levels
of antioxidant enzymes and vitamin.
Keywords Oxidative stress Glutathione peroxidase
Ovariectomize Hypercholesterolemia Antioxidant
vitamins Liver
ACV Apple cider vinegar
GSH Glutathione
GSH-Px Glutathione peroxidase
HDL High-density lipoprotein
i.p. Intraperitoneal
LDL Low-density lipoprotein
OVX Ovariectomized
ROS Reactive oxygen species
VLDL Very low-density lipoprotein
Diseases of liver and kidney abnormalities constitute a
serious health problem in menopause (Poli 1993). These
diseases are usually treated with drugs, dietary or metabolic
changes, and vaccines. Liver disease in menopause also
induces lipid profile abnormalities because of its important
role in lipid metabolism (Luotola et al. 1986; Ulas¸ and Cay
2011). In this regard, finding new, safe, and effective food
supplements has become an important line of research.
Oxidative stress is the result of an imbalance between
the rates of free radical production and elimination via
endogenous antioxidant mechanisms such as the enzymes
glutathione peroxidase (GSH-Px) and catalase as well as
M. Nazırog
˘lu (&)M. Ku
¨kayaz E. So
Department of Biophysics, Medical Faculty, Suleyman Demirel
University, Isparta, Turkey
M. Gu
¨ler G. Saydam
Suleyman Demirel University, Isparta, Turkey
C. O
Regenerative and Restorative Medical Research Center,
Istanbul Medipol University, Istanbul, Turkey
J Membrane Biol (2014) 247:667–673
DOI 10.1007/s00232-014-9685-5
low molecular weight reductants like a-tocopherol, gluta-
thione (GSH), and ascorbate (Kovacic and Somanathan
2008). This imbalance can be initiated by numerous factors
including acidosis, transition metals, nitric oxide, LDL-
oxidation, and uncouplers of mitochondrial electron
transport (Nazırog
˘lu and Bransch 2006; Espino et al. 2012).
The major roles of GSH-Px are to catalyze the reduction
of hydrogen peroxide to water (Kovacic and Somanathan
2008) and the removal of organic hydroperoxides (Nazır-
˘lu 2009). GSH is a peptide involved in maintaining
oxidant homeostasis and in the cellular detoxification of
reactive oxygen species (ROS) in tissues including brain,
kidney, and liver (Halliwell 2006). Vitamin E is the most
important lipid antioxidant in cells. In addition to its role as
free radical scavenger, vitamin C also transforms vitamin E
to its active form (Frei et al. 1989).
It has been reported that a number of phenolic compound
extracted from fruit can protect against oxidative stress-
induced liver and kidney injury because of their antioxidant
properties (Yang et al. 2010;Nazırog
˘lu et al. 2011a,b). This
has promoteda substantial increase in the use of supplemental
dietary phenols and alternative therapy to treat menopause-
induced liver and kidney diseases in women and animals in
favor of hormone replacement treatment (Nazırog
˘lu et al.
˘lu et al. 2011a,b).
Apple cider vinegar is widely used in salad dressings,
marinades, vinaigrettes, food preservatives, chutneys, and
other common foods. The main classes of phenols in apples
and ACV are flavonoids and polyphenolic compounds (Denis
et al. 2013). A number of studies have shown that these foods
have a variety of pharmacological functions, including anti-
oxidant (Yang et al. 2010; Denis et al. 2013), antidiabetic
(Shishehbor et al. 2008), and cholesterol lowering (Budak
et al. 2011) properties, without adverse effects. However,
information on the effects of ACV on antioxidant systems is
very limited. For example, there are no scientific reports on the
use of ACV in menopausal women or ovariectomized animals
as a potential source of natural antioxidant phenols.
It is known, however, that the deficiency of ovarian hor-
mones promotes the generation of ROS, resulting in oxidative
stress, cell damage, or death (Budak et al. 2011). The bene-
ficial effects of estrogen include reducing total- and low-
density cholesterol (LDL) through enhanced LDL receptor
binding and clearance (Noh et al. 1999; Sanchez-Rodriguez
et al. 2011). Additionally, by reducing hepatic lipase activity,
estrogen promotes the formation of larger, less atherogenic
LDL (Nazırog
˘lu et al. 2004a,b; Sanchez-Rodriguez et al.
2011). Hence, the use of ACV instead of cholesterol lowering
drug therapy in menopause may improve oxidative stress-
induced lipid profile liver and kidney function in ovariecto-
mized mice as a model of postmenopausal women.
The aim of the current study was to investigate protec-
tive effects of ACV on ovariectomy-induced erythrocyte,
liver, and kidney oxidative damage as well as lipid profile
changes in mice fed high cholesterol and its free radical
scavenging activity in vivo.
Materials and Methods
Forty female Swiss mice weighing 36–40 g were used for the
experimental procedures. Twenty of them remained intact as
control and ACV groups, and the ovaries were removed in
the remaining 20 animals in order to constitute groups II and
IV. The mice were housed in individual plastic cages with
bedding. Standard food and tap water were available
ad libitum for the duration of the experiments unless other-
wise noted. The temperature was maintained at 22 ±2°C.
A 12/12 h light/dark cycle was maintained, unless otherwise
noted. The ACV was provided as a gift from the Agricultural
Faculty of Suleyman Demirel University and it was diluted
with water so that it would provide 0.6 % of the animals’
daily diet (Budak et al. 2011). This concentration was found
to be innocuous to mice in a pre-study in which various
concentrations of ACV were tested. Hypercholesterolemia
was induced by daily gavage administration of 1 ml/100 g
body weight of a cocktail containing 100 g cholesterol, 30 g
propylthiouracil, and 100 g cholic acid in 1 liter peanut oil
(Vogel and Vogel 1997).
The mice were maintained and used in accordance with
the Animal Welfare Act for the Care and Use of Laboratory
Animals of Suleyman Demirel University (SDU), and the
local ethical committee of the Medical Faculty, SDU,
approved the experimental protocol designed for this study
(Protocol Number; 2009: 27-08).
Experimental Groups
The mice were randomly divided into four groups of ten
that were treated for 28 days, as follows:
Group I was used as control and was fed a cholesterol-
rich feed (5 % cholesterol). The only treatment was intra-
gastrical application of physiological saline, as placebo
(Dilek et al. 2010).
Group II (OVX), the ovaries were surgically removed.
Otherwise, the mice were treated as described for the
Group III (ACV) for which in addition to the high-
cholesterol diet, the animals were given 0.6 % apple cider
vinegar instead of a placebo.
Group IV (ACV ?OVX) in which the animals were
ovariectomized and treated with ACV, as described for
group III.
668 M. Nazırog
˘lu et al.: Ovariectomy and Hypercholesterolemia
Animal Model of Menopause
For removal of the ovaries, the two OVX groups of mice
were anesthetized by intraperitoneal administration of a
cocktail of ketamine hydrochloride (50 mg/kg) and xylazine
(5 mg/kg) (Kireev et al. 2010; Dilek et al. 2010). Briefly, the
surgical procedure for ovariectomy equithesin and two
4-mm incisions were made through the skin and the muscle
back walls in parallel with the bodyline. The ovaries were
then located, and a silk thread was tightly tied around the
oviduct, including the ovarian blood vessels. The wound was
sutured with a synthetic absorbable thread. Depocilin was
used to prevent of infection. The ovariectomized mice were
allowed to fully recover before beginning the study protocol.
Blood and Tissue Samples
All the mice were sacrificed after 12 h of the last ACV
administration to obtain the necessary blood and tissue sam-
ples. The blood was separated into plasma and erythrocytes by
centrifugation at 1,5009gfor 10 min at ?4°C. The eryth-
rocyte samples were washed three timesin cold isotonic saline
(0.9 %, v/w), and hemolysis was accomplished by adding a
nine-fold volume of 50 mM, pH 7.4 phosphate buffer. The
hemolyzed samples were stored at -30 °C for not more than
months pending measurement of enzymatic activity.
All preparation procedures were performed on ice. The
liver and kidney samples were washed twice with cold
saline solution, placed into glass bottles, labeled, and stored
in at -33 °C until needed. After weighing, the tissue
samples were placed on ice, cut into small pieces with
scissors and mixed with five volumes (1:5 w/v) of ice cold
50 mM, pH 7.4 Tris-HCl buffer, and then homogenized for
2 min at 5,000 rpm using a glass Teflon homogenizer
(Caliskan Cam Teknik, Ankara, Turkey). The homogenate
was immediately used for establishing the extent of lipid
peroxidation and antioxidant enzyme levels. The antioxi-
dant vitamin analyses were performed within 3 months.
Lipid Peroxidation Determination
The lipid peroxidation levels were determined by the TBA
method of Placer et al. (1966) using a UV-1800 Spectrom-
eter (Schimadzu, Kyoto, Japan). The values are expressed as
lmol/gram protein and as lmol/gram hemoglobin (Hb).
Reduced Glutathione (GSH), Glutathione
Peroxidase (GSH-Px), and Protein Assays
The GSH content in erythrocyte, kidney, and liver samples
was measured by the spectroscopic method of Sedlak and
Lindsay (1968), as described in a previous study (Nazır-
˘lu et al. 2004a). The GSH-Px activity in erythrocyte,
kidney, and liver was measured spectrophotometrically at
37 °C according to the method of Lawrence and Burk
(1976). The protein contents in the liver and kidney were
measured by the method of Lowry et al. (1951) with bovine
serum albumin as the standard. Drabkin’s reagent was used
in the erythrocyte for determination of Hb.
Vitamins A, C, and E and b-carotene Analyses
The levels of vitamins A and E were determined in the
kidney and liver samples by a modification of the methods
described by Desai (1984) and Suzuki and Katoh (1990)as
described in a previous study (Nazırog
˘lu et al. 2004a).
Liver and kidney samples (0.25 g) were saponified by the
addition of 0.3 ml KOH (60 % w/v in water) and 2 ml of
1 % (w/v in ethanol) ascorbic acid, followed by heating at
70 °C for 30 min. After cooling on ice, 2 ml of water and
1 ml of n-hexane were added to the samples, mixed, and
allowed to separate into phases. An aliquot of 0.5 ml of
n-hexane extract was taken, and the vitamin A concentra-
tions were measured at 325 nm. Then, reactants were
added and the absorbance value of hexane was measured in
a spectrophotometer at 535 nm. Calibration was performed
using standard solutions of all-trans retinol and a-tocoph-
erol in hexane.
The concentrations of b-carotene in the kidney and liver
samples were determined according to the method of Su-
zuki and Katoh (1990). 2 ml of hexane was mixed with
0.25 g tissue sample. The concentration of b-carotene in
hexane was measured spectrophotometrically at 453 nm.
The quantitative determination of ascorbic acid in the
kidney and liver samples was performed according to the
method of Jagota and Dani (1982). The absorbance of the
samples was measured spectrophotometrically at 760 nm.
Measurement of Triglyceride and Total Cholesterol
Serum triglycerides, total cholesterol, VLDL cholesterol,
and element levels were measured in an autoanalyzer
(Olympus AV 2700) at the Isparta State Hospital by using
standard laboratory techniques (Nazırog
˘lu et al. 2011a,b).
Statistical Analysis
All results were expressed as mean ±SD. Significant
values in the four groups were assessed by the unpaired
Mann–Whitney Utest. Data were analyzed using the SPSS
statistical software (v. 17.0, SPSS Inc. Chicago, Illinois,
USA). pvalues of less than 0.05 were regarded as
M. Nazırog
˘lu et al.: Ovariectomy and Hypercholesterolemia 669
Serum Element, Total Cholesterol and Triglycerides
Serum total cholesterol, triglycerides, and VLDL choles-
terol levels are shown in Figs. 1and 2, respectively. The
total cholesterol levels as in groups I–IV were 113, 121, 79,
and 89 mg/dl, respectively. The triglycerides levels in
these groups were 111, 142, 78, and 73 mg/dl, respectively.
In the same order, the total cholesterol levels were 22, 43,
16, and 20 mg/dl, respectively. The serum triglycerides
(p\0.05), total (p\0.05) and VLDL (p\0.001) cho-
lesterol levels were significantly higher in group II relative
to controls, but significantly lower in groups III and IV
relative to groups I and II (p\0.001). The levels of
sodium, potassium, and chlorine in serum samples are
shown in Table 1. No significant changes in the levels of
these elements were detected in all study groups.
Lipid Peroxidation Results
The lipid peroxidation levels in erythrocytes, liver, and
kidney are shown in Tables 2,3, and 4, respectively. Lipid
peroxidation is considered to be one of the principal indi-
cators of OVX-induced oxidative erythrocytes, liver, and
kidney injury (Nazırog
˘lu et al. 2004a,b; Ulas¸ and Cay
2011; Sanchez-Rodriguez et al. 2011). The lipid peroxi-
dation levels in the erythrocytes (p\0.05), liver
(p\0.05), and kidney (p\0.01) were significantly higher
in group II than in the controls. However, pretreatment of
mice with ACV effectively inhibited OVX-induced lipid
peroxidation levels in l erythrocytes (p\0.05), liver
(p\0.05) and kidney (p\0.01).
GSH and GSH-Px Values in Liver and Kidney
GSH-Px is a strong antioxidant enzyme within enzymatic
ROS scavenger systems, while GSH is a non-enzymatic
thiol-containing antioxidant against OVX-induced cellular
toxicity (Nazırog
˘lu 2009; Kovacic and Somanathan 2008).
The GSH levels and GSH-Px activity in erythrocytes, liver,
and kidney are shown in Tables 2,3, and 4, respectively.
The data show that the GSH-Px activity in erythrocytes
(p\0.05) and kidney (p\0.01), and GSH levels in
erythrocytes (p\0.05) significantly decreased in group II.
However, the GSH-Px activity in erythrocytes (p\0.05)
and kidney (p\0.01), and the GSH levels in erythrocytes
(p\0.05) were higher in groups III and IV relative to
group II. The GSH level in liver and kidney and GSH-Px
activity in liver did not change in the four groups
Antioxidant Vitamin Concentrations in Liver
and Kidney
The concentrations of b-carotene and the levels of vitamins
A, C, and E in liver and kidney are shown in Tables 3and
4, respectively. The vitamin E stores in liver were mark-
edly depleted in group II (p\0.05), and the vitamin E and
b-carotene concentrations in liver were significantly
depleted in groups III and IV (p\0.01) relative to groups
II and I.
Pretreatment of mice with ACV effectively increased
the levels of vitamins C (p\0.05), E (p\0.05), and
b-carotene (p\0.01) in kidney. The vitamin A concen-
trations in liver and kidney did not significantly change in
any of the four groups.
It was observed that the lipid peroxidation values for
erythrocytes, liver, and kidney and the serum triglycerides,
total and VLDL cholesterol concentrations were increased
and that the GSH-Px activity and level of GSH were
decreased in ovariectomized mice, proving that the proto-
col worked as an experimental menopause model. Sup-
plementation with ACV resulted in an increase of the
Fig. 1 Effects of apple cider vinegar (ACV) on total cholesterol and
triglyceride levels in serum of ovariectomized (OVX) mice
(mean ±SD and n=10).
p\0.05 and
p\0.001 as compared
with group control.
p\0.001 as compared with OVX group
Fig. 2 Effects of apple cider vinegar (ACV) on VLDL cholesterol
levels in serum of ovariectomized (OVX) mice (mean ±SD and
p\0.001 p\0.001 versus control.
p\0.001 versus
OVX group
670 M. Nazırog
˘lu et al.: Ovariectomy and Hypercholesterolemia
antioxidants GSH and GSH-Px and a decrease of the tis-
sues lipid peroxidation and serum lipid profiles.
Lipid peroxidation has been implicated in the patho-
genesis of OVX-induced oxidative injury due to meno-
pause-induced estrogen deficiency, which results in cell
membrane damage. Sexual hormones play an important
role in the progression of liver diseases. The lipid
peroxidation level was higher in the OVX mice, revealing a
deficient antioxidant defense system. This observation
confirms previous studies showing that OVX led to an
increase of lipid peroxidation and decrease of GSH-Px
activity and antioxidant vitamin levels (Poli et al. 1993;
˘lu et al. 2011a,b; Nazırog
˘lu et al. 2004a,b; Dilek
et al. 2010; Lucas et al. 2006).
Table 1 Effects of apple cider
vinegar (ACV) supplementation
on serum sodium, potassium,
and chloride levels in
ovariectomized (OVX) mice
(mean ±SD)
Parameters Control (n=10) OVX (n=10) ACV (n=10) OVX ?ACV (n=10)
Sodium (mg/dl) 163.2 ±24.8 145.2 ±9.2 142.5 ±7.4 155.5 ±21.2
Potassium (mg/dl) 4.60 ±0.89 4.91 ±0.81 4.70 ±0.40 4.68 ±0.59
Chloride (mg/dl) 124.8 ±16.7 110.2 ±4.9 107.0 ±7.8 113.3 ±14.8
Table 2 Effects of apple cider vinegar (ACV) supplementation on erythrocytes lipid peroxidation (LP), reduced glutathione (GSH), and
glutathione peroxidase levels in ovariectomized (OVX) mice (mean ±SD)
Parameters Control (n=10) OVX (n=10) ACV (n=10) OVX ?ACV (n=10)
GSH-Px (IU/g Hb) 13.00 ±1.49 10.90 ±3.52
12.80 ±3.07
12.90 ±2.90
GSH (lmol/g Hb) 10.8 ±1.28 9.37 ±1.55 10.40 ±1.73 10.20 ±1.51
LP (lmol/g Hb) 11.80 ±1.44 14.30 ±1.31
12.00 ±1.85
11.70 ±1.42
p\0.05 versus control group
p\0.05 versus OVX group
Table 3 Effects of apple cider vinegar (ACV) supplementation on liver lipid peroxidation (LP), reduced glutathione (GSH), glutathione
peroxidase (GSH-Px), vitamin A, C, E and b-carotene levels in ovariectomized (OVX) mice. (mean ±SD)
Parameters Control (n=10) OVX (n=10) ACV (n=10) OVX ?ACV (n=10)
GSH-Px (IU/g protein) 16.90 ±1.39 17.70 ±2.06 16.50 ±3.32 16.30 ±3.52
GSH (lmol/g protein) 6.93 ±0.87 6.56 ±0.39 7.55 ±1.31 6.86 ±1.58
LP (lmol/g protein) 10.80 ±1.26 13.30 ±1.59
9.86 ±1.52
10.70 ±1.00
Vitamin A (lmol/g tissue) 50.16 ±0.45 53.89 ±1.84 49.84 ±1.17 53.07 ±0.95
b-carotene (lmol/g tissue) 2.62 ±0.11 2.35 ±0.09 2.06 ±0.39
1.84 ±0.17
Vitamin C (lmol/g tissue) 0.68 ±0.09 0.66 ±0.14 0.63 ±0.09 0.69 ±0.08
Vitamin E (lmol/g tissue) 9.54 ±0.56 9.10 ±0.46
7.62 ±1.80
6.08 ±0.96
p\0.05 and
p\0.01 versus control group
p\0.05 and
p\0.01 versus OVX group
Table 4 Effects of apple cider vinegar (ACV) supplementation on kidney lipid peroxidation (LP), reduced glutathione (GSH), glutathione
peroxidase (GSH-Px), vitamin A, C, E and b-carotene levels in ovariectomized (OVX) mice (mean ±SD)
Parameters Control (n=10) OVX (n=10) ACV (n=10) OVX ?ACV (n=10)
GSH-Px (IU/g protein) 14.70 ±0.83 12.80 ±1.21
15.90 ±1.12
14.88 ±1.88
GSH (lmol/g protein) 5.07 ±0.39 5.26 ±0.29 5.07 ±0.47 5.26 ±0.73
LP (lmol/g protein) 4.60 ±0.89 6.11 ±1.05
4.93 ±0.48
6.34 ±0.78
Vitamin A (lmol/g tissue) 10.47 ±1.97 10.37 ±1.26 12.01 ±1.38 11.13 ±1.54
b-Carotene (lmol/g tissue) 0.81 ±0.07 0.79 ±0.09 1.06 ±0.17
1.83 ±0.13
Vitamin C (lmol/g tissue) 0.35 ±0.09 0.34 ±0.09 0.50 ±0.07
0.50 ±0.06
Vitamin E (lmol/g tissue) 6.72 ±0.67 6.17 ±0.71 7.62 ±1.80
7.65 ±0.99
p\0.05 and
p\0.01 versus control group
p\0.01 and
p\0.001 versus OVX group
M. Nazırog
˘lu et al.: Ovariectomy and Hypercholesterolemia 671
It is well known that menopause and removal of ovaries
induce kidney and liver oxidative injury (Ulas¸ and Cay 2011;
Barp et al. 2012). Additionally, the present study was also
designed to explore the protective effects of ACV as free
radical scavenger on OVX-induced oxidative damage to
erythrocyte, liver, and kidney tissues. Administration of ACV
improved the antioxidant defense system and decreased lipid
peroxidation in erythrocytes, kidney, and liver.
Flavonoids and phenolic agents occur widely in many
plants. Recently, the role of phenolic compound and
flavonoids in the prevention of menopause-induced oxi-
dative stress and disease has gained great interest (Yang
et al. 2010; Nazırog
˘lu et al. 2011a,b; Nikolic
´et al. 2012).
Apple cider vinegar contains high concentrations of poly-
phenols and flavonoids (Yang et al. 2010; Budak et al.
2011; Denis et al. 2013), which explains its antioxidant
properties against oxidative damage to erythrocytes and
In mammalians, the combined actions of various cellular
antioxidants are critical for the effective detoxification of
free oxygen radicals. Among the cellular antioxidant
enzymes, GSH-Px has been extensively studied. It converts
hydrogen peroxide to water by using GSH as substrate
(Kovacic and Somanathan 2008 Nazırog
˘lu 2009 ). Ovari-
ectomized mice showed decreased antioxidant capacity in
kidney and erythrocytes, as evidenced by decreased
activity of the antioxidant enzymes, which is in agreement
with earlier apple and apple juice reports (Poli 1993;
˘lu et al. 2004a,b; Avci et al. 2007; Dilek et al.
2010; Nazırog
˘lu et al. 2011a,b). Treatment with ACV
prevented the reduction of the antioxidant enzyme activity
and subsequent oxidative injury to erythrocytes and kidney
(Avci et al. 2007; Ulas¸ and Cay 2011; Yuan et al. 2011).
Vitamin C has been shown to be an important antioxi-
dant, to regenerate vitamin E through redox cycling, and to
raise intracellular GSH levels (Frei et al. 1989). As such,
vitamin C plays an important role in protein -SH group
protection against oxidation. High concentrations of vita-
min C have are found in ACV (Denis et al. 2013). The
GSH-Px activity, vitamin C, vitamin E, and b-carotene
concentrations in kidney, and GSH-Px and GSH level in
erythrocytes increased in the ACV-treated groups. These
observed increases indicate an important role of ACV in
normalizing GSH-Px and antioxidant vitamin concentra-
tion in OVX mice.
The vitamin E and total cholesterol concentrations in
liver were also decreased in the ACV- and OVX ?ACV
groups. Vitamin E is an important antioxidant in biological
systems (Nazırog
˘lu 2007). Vitamin E promotes homeo-
stasis in living cells by a mechanism of incorporation into
cell membranes or by entering the cells. Liver possesses an
a-tocopherol-binding protein specific for vitamin E, facil-
itating its incorporation into cells and subsequent transfer
from liver to cells through cholesterols (Hacquebard and
Carpentier 2005; Traber 2007). The property of tocopherol
that appears to be related to most manifestations of defi-
ciency is its inhibitory effect on the auto-oxidation of
unsaturated fatty acids.
Estrogen deficiency increases generation of free oxygen
radicals, which induces oxidative stress and results in cell
damage or death (Budak et al. 2011). Beneficial effects of
estrogen in liver and blood include reducing cholesterol
through enhanced LDL receptor binding and clearance
(Noh et al. 1999; Sanchez-Rodriguez et al. 2011). Addi-
tionally, by reducing hepatic lipase activity, treatment with
this hormone promotes the formation of larger and less
atherogenic LDL cholesterol particles (Noh et al. 1999;
˘lu et al. 2004a,b).
In the present study, we were not able to measure all
serum lipid values such as HDL and LDL values in mice
due to limited amounts of serum. However, we observed
that OVX-induced changes of triglycerides, total and
VLDL cholesterol are modulated by ACV supplementa-
tion. Similarly, Shishehbor et al. reported that ACV
improved the serum lipid profile in normal and diabetic rats
by decreasing serum triglycerides, LDL and increasing
serum HDL-cholesterol (Shishehbor et al. 2008). It was
also recently reported that ACV decreased triglyceride and
VLDL levels in rats fed high cholesterol when compared to
animals on high-cholesterol diets without ACV supple-
mentation (Budak et al. 2011). In agreement with these
observations, total cholesterol and triglycerides were lower
in the ACV-treated mice fed high cholesterol.
In conclusion, the results presented in this study suggest
that ovariectomy is associated with an increase of serum
triglycerides, total and VLDL cholesterol and with eryth-
rocyte, liver, and kidney lipid peroxidation and reduction
of some antioxidants in tissues and cells. The administra-
tion of apple cider vinegar possesses a protective effect
against ovariectomy-induced blood, liver, and kidney oxi-
dative injury in mice fed high cholesterol and that such
protection may be due to free oxygen radical scavenging
effects, reduced lipids and lipid peroxidation, and increased
antioxidant enzyme and vitamin levels. The results in
blood, liver, and kidney may be of help to physicians and
nutritionists in the use of apple cider vinegar as supplement
in the treatment of oxidative stress-induced toxicity.
Acknowledgments MN formulated the present hypothesis and was
responsible for writing the report. MG, GS, CO
¨, MK, and ES were
responsible for analysis of the data. The authors wish thanks to Dr.
Manuel Flores-Arce (Tijuana University, Mexico) for polishing
English of the manuscript. The study was partially supported by
Scientific Turkish Scientific and Technical Research Institute
Conflict of interest The authors declare that there are no conflicts
of interest in the current study.
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... Additionally, fruit by-products such as apple vinegar have been documented as having several biological properties due to its rich composition in phenolic compounds [30][31][32][33][34][35][36][37][38]. Apple vinegar is considered to be an antioxidant, antihyperglyceamic, antihyperlipidimic and hepato-nephroprotective agent [39][40][41][42]. Apple vinegar contains a large group of bioactive compounds with high antioxidant potential, which counteract reactive oxygen species (ROS) [43]. ...
... A study published by [32] revealed that apple vinegar enhanced the antioxidant defense system, which could decrease enzyme leakage into plasma. The same findings were evoked by [41]. In relation to our previous studies, recently, we proved that apple vinegar exhibited an important antioxidant effect in vivo against hydrogen peroxide at the dose of 10 mL/kg b.w during 22 days of treatment by reducing liver and kidney damage and decreasing hepatic enzymes and creatinine [38]. ...
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This study aims to examine the ability of apple vinegar on phenylhydrazine (PHZ)-induced hemolytic anemia in Wistar rats. In vitro, phenolic and flavonoid content and antioxidant activity were determined. In vivo, phenylhydrazine (10 mg/kg) was injected intravenously into rats for 4 days and then treated with apple vinegar daily by gavage (1 mL/kg) for five weeks. high level of polyphenols and flavonoids (90 ± 1.66 mg GAE/100 mL and 7.29 ± 0.23 mg QE/100 mL, respectively) were found in the apple vinegar which gives it a good ability to scavenge free radicals (TAC = 4.22 ± 0.18 mg AAE/100 mL and DPPH, IC50= 0.49 ± 0.004 µL/mL). The phytochemical composition of apple vinegar revealed the presence of numerous bioactive compounds including arbutin, apig-enin, sinapic, ferulic and trans-ferulic acids. The major antioxidant components in apple vinegar were ferulic and trans-ferulic acids (40% and 43%, respectively). PHZ treatment induced changes in platelets, blood cell count, mean corpuscular volume, hemoglobin concentration and mean capsu-lated hemoglobin. However, the co-administration of apple vinegar revealed its capacity to ameliorate the changes induced by phenylhydrazine. Therefore, apple vinegar use could have a positive impact on the prevention of hemolytic anemia induced by phenylhydrazine due to the antioxidant properties of its major components.
... The main component of ACV is acetic acid which majorly confers on the vinegar the beneficial effect it possesses (Kondo et al. 2009;Abdulrauf et al. 2018). Apple cider vinegar has been extensively reported as having protective effect against erythrocyte, kidney and liver injuries (Naziroǧlu et al. 2014), acting as antidyslipidemic agent (Beheshti et al. 2012;Halima et al. 2018), and possessing antioxidant activity (Xia et al. 2020). It is also commonly used as antidiabetic, antiobesogenic and antihypertensive agent (Johnston and Buller 2005;Bouderbala et al. 2015). ...
... It has been reported severally that ACV acts upon body fat which is a key factor in the reduction of body weight (Hamed and Matar 2014;Naziroǧlu et al. 2014). The body mass index (BMI) being relatively lower in all the treated groups may be attributed to this factor. ...
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Apple cider vinegar (ACV) is used globally as food supplement and widely acclaimed as therapeutic agent for various ill-health conditions. This study investigated the effect of consuming ACV during pregnancy on the haematological and biochemical profile of adult female albino rats in correlation with morphometric of the litters. Graded concentrations of ACV were administered to gestating albino rats to evaluate their effects on haematological and biochemical parameters of gestating rats and morphology of their pups at delivery. Ninety-six rats comprising 48 females (12 to 13 weeks old) and 48 males (15 weeks old) were used. The impregnated rats were randomly assigned to four groups (A, B, C and D) of 12 rats each. Each group was further sub-assigned into three replicates of four rats. Group A served as the untreated control while groups B, C and D were treated. Before and after 3-week period of treatment, blood samples were taken from the rats for haematological and serum biochemical evaluations following standard procedures. There were no significant variations (p > 0.05) between the groups in pup litter size, crown rump length and body mass index (BMI). There were also no significant variations (p > 0.05) between the groups in post-treatment values of packed cell volume, haemoglobin concentration, red blood cell count, absolute eosinophil counts and serum levels of calcium. Lower pup weight and higher litter size (at lower doses), lower lymphocyte and neutrophil counts, and lower serum activity of ALT, AST and ALP enzymes were recorded. ACV as used in this study has no adverse effects on haematological and biochemical profiles of gestating rats and their pups. It may therefore be safe to take ACV during pregnancy apart from other uninvestigated consequences.
... 17 These results were in line with Nazıroğlu et al. 2014 who observed in their study that triglycerides and cholesterol were lesser among mice treated with ACV and fed high cholesterol. 18 Cinnamon also has hypocholesterolemic properties which were proven by a study conducted by Alsoodeeri et al. on hypercholesterolemic rats. It was found in the study that cinnamon extract given as a daily dose of 4g/kg presented antihyperlipidemic effect among hypercholesterolemic rats. ...
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Introduction: Metabolic syndrome is constituted by hyperlipidemia with presentation of diverse lipid profile including hypertriglyceridemia, hypercholesterolemia and hereditary combined hyperlipidemia. Objectives: To find the synergistic effect Cinnamomum cassia and apple cider vinegar on combined administration in albino mice. Methodology: This was an experimental study and conducted at Department of Pharmacology, Al-Nafees Medical College and Hospital Islamabad in collaboration with Animal House of the National Institute of Health, Islamabad-Pakistan. A total of 50 adult albino mice were randomly divided into 5 groups. Hyperlipidemia was induced during the first two weeks of experimentation, and this was followed by treatment period of 6 weeks. Groups were categorized as A (Normal Control), B (Treated Control), C (Treatment Group I), D (Treatment Group II), and E (Treatment Group III). Diet pattern for these groups remained to be high cholesterol, Simvastatin 0.6 mg/Kg BW, ACV upto 15% of animal feed, Cinnamon powder 6mg/Kg BW and Cinnamon with high cholesterol diet respectively. Blood samples to estimate parameters of lipid profile were collected at 0,15th, 30th, 45th and 60th days. Results: A significant decrease (p<0.05) was observed in total cholesterol, triglyceride and low density lipoprotein at 45th and 60th days as compared to normal control group. Serum high density lipoprotein was low at 15th day among all groups because of induction of hypercholesterolemia, which was then noted to be reversed at 60th day. Highest surge was noted to be in group E and significant increase in all groups (p<0.05) was noted in all groups as compared to normal control. Conclusion: Apple Cider Vinegar keeps the anti-hyperlipidemia effect which was found to be significantly high (p<0.05) in combination with Cinnamomum cassia when consumed by male albino mice. Key Words: Apple cider vinegar, Cinnamon, Lipid profile, Albino mice, Metabolic syndrome, Lipoprotein
... *P > 0.05 nonsignificant relative to control; # P < 0.05 significant relative to control; and @ P < 0.05 significant relative to HgCl 2 group with its major constituents as flavonoids and polyphenols provide several pharmacological actions, including antioxidant properties; so could reverse the oxidative stress on hepatocytes, consequently decreasing the cellular levels of HSP60. [26] conclusIon ACV could attenuate HgCl 2 -induced ultrastructural and immunohistochemical changes in rat hepatocytes through its strong antioxidant properties. Hence, ACV could be promising for attenuation of liver cell damages induced by various toxins. ...
... Moreover, it is suggested by literature and clinical reports that a proper diet including vitamins, proteins, and minerals will surely complement the medicine for the treatment of AD [89][90][91]. Apple cider has been reported to increase the activity of SOD, CAT, and glutathione peroxidase (GPx) to reduce lipid peroxidation [92,93]. ...
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Alzheimer’s disease (AD) rate is accelerating with the increasing aging of the world’s population. The World Health Organization (WHO) stated AD as a global health priority. According to the WHO report, around 82 million people in 2030 and 152 million in 2050 will develop dementia (AD contributes 60% to 70% of cases), considering the current scenario. AD is the most common neurodegenerative disease, intensifying impairments in cognition, behavior, and memory. Histopathological AD variations include extracellular senile plaques’ formation, tangling of intracellular neurofibrils, and synaptic and neuronal loss in the brain. Multiple evidence directly indicates that oxidative stress participates in an early phase of AD before cytopathology. Moreover, oxidative stress is induced by almost all misfolded protein lumps like α-synuclein, amyloid-β, and others. Oxidative stress plays a crucial role in activating and causing various cell signaling pathways that result in lesion formations of toxic substances, which foster the development of the disease. Antioxidants are widely preferred to combat oxidative stress, and those derived from natural sources, which are often incorporated into dietary habits, can play an important role in delaying the onset as well as reducing the progression of AD. However, this approach has not been extensively explored yet. Moreover, there has been growing evidence that a combination of antioxidants in conjugation with a nutrient-rich diet might be more effective in tackling AD pathogenesis. Thus, considering the above-stated fact, this comprehensive review aims to elaborate on the basics of AD and antioxidants, including the vitality of antioxidants in AD. Moreover, this review may help researchers to develop effectively and potentially improved antioxidant therapeutic strategies for this disease as it also deals with the clinical trials in the stated field.
... Previous studies reported that apple vinegar exhibited a remarkable antioxidant activity [24][25][26]. It has been proved that apple vinegar has several pharmacological properties including antidiabetic effect, antibacterial effect, antihypertensive effect, and anti-inflammatory effect [25,[27][28][29][30][31][32]. ...
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The main objective of the current study was to determine the physicochemical properties, antioxidant activities, and α-glucosidase and α-amylase inhibition of apple vinegar produced by artisanal and industrial methods. Apple vinegar samples were analyzed to identify their electrical conductivity, pH, titratable acidity, total dry matter, Brix, density, mineral elements, polyphenols, flavonoids, and vitamin C. The antioxidant activity of apple vinegar samples was evaluated using two tests, total antioxidant capacity (TAC) and DPPH radical scavenging activity. Finally, we determined α-glucosidase and α-amylase inhibitory activities of artisanal and industrial apple vinegar. The results showed the following values: pH (3.69–3.19); electrical conductivity (2.81–2.79 mS/cm); titratable acidity (3.6–5.4); ash (4.61–2.90); ◦Brix (6.37–5.2); density (1.02476–1.02012), respectively, for artisanal apple vinegar and industrial apple vinegar. Concerning mineral elements, potassium was the most predominant element followed by sodium, magnesium, and calcium. Concerning bioactive compounds (polyphenols, flavonoids, and vitamin C), the apple vinegar produced by the artisanal method was the richest sample in terms of bioactive compounds and had the highest α-glucosidase and α-amylase inhibition. The findings of this study showed that the quality and biological activities of artisanal apple vinegar were more important than industrial apple vinegar
... In our previous studies, we noticed that apple cider vinegar can ameliorate the metabolic disorder induced by a hypercaloric diet, and it possesses antibacterial and antifungal activities [18][19][20]. Recent researchers have explored that vinegar exhibited antioxidative properties by decreasing ROS accumulation and upregulating the expression of antioxidative enzymes [21][22][23]. It has been shown that vinegar administration exhibits a potent hepatoprotective effect against oxidative damages induced by hydrogen peroxide [24]. ...
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Oxidative stress is the trigger of several diseases. It is an imbalance between the production of free radicals and antioxidants. This study aims to evaluate the antioxidant capacity and the protective property of Rosa canina fruits and apple cider vinegar combined or not against hydrogen peroxide (H2O2)-induced toxicity in Wistar rats. The experiment included five groups: group 1 received distilled water (10 mL/kg b.wt), group 2 received H2O2 10% (10 mL/kg b.wt), group 3 received H2O2 10% (10 mL/kg b.wt) and apple vinegar (2 mL/kg b.wt); group 4 received H2O2 10% (10 mL/kg b.wt) and apple vinegar supplemented with Rosa canina fruits extract (300 mg/kg b.wt); group 5 received H2O2 10% (10 mL/kg b.wt) and extract of Rosa canina fruits (300 mg/kg b.wt). The doses were given once daily via a gavage. The antioxidant capacity of apple vinegar and Rosa canina extract was analyzed, and AST, ALT, PAL, urea, and creatinine were determined on day 22 of the experiment. In addition, the kidney and the liver tissues were analyzed. The results showed that H2O2 caused a significant elevation of blood urea, blood creatinine, and transaminases. The histopathology examination revealed that H2O2 caused congestion, hemorrhage, and Bowman’s space enlarged. On the other hand, the results clearly showed that apple vinegar and Rosa canina fruits counterbalance the biochemical and histological changes induced by H2O2. In conclusion, the two natural products studied in this work are effective against the harmful effect of oxidative stress, which explains their use in traditional medicine.
... Acetic acid as the main organic acid in vinegars is the key factor of numerous beneficial properties through the activation of the MAPK pathway, which induces reduction of blood glucose, increases glycogen storage, reduces triglycerides levels, increases insulin sensitivity, and decreases insulin resistance [18]. The aforementioned features revealed that acetic acid is the main acid of vinegar that can ameliorate metabolic disorders and ameliorate disease markers [16,18,22,[63][64][65][66][67][68][69][70][71]. ...
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The popularity of fruits vinegar (FsV) has been increased recently as a healthy drink wealthy in bioactive compounds that provide several beneficial properties. This review was designed in the frame of valorization of fruits vinegar as a by-product with high value added by providing overall information on its biochemical constituents and beneficial potencies. It contains a cocktail of bioactive ingredients including polyphenolic acids, organic acids, tetramethylperazine, and melanoidins. Acetic acid is the most abundant organic acid and chlorogenic acid is the major phenol in apple vinegar. The administration of fruits vinegar could prevent diabetes, hypercholesterolemia, oxidative stress, cancer, and boost immunity as well as provide a remarkable antioxidant ability. The production techniques influence the quality of vinegar, and consequently, its health benefits.
In this work, a colorimetric sensor array (CSA) for quantitative determination of total acids in apple vinegar during fermentation was constructed. The sensor array was properly designed based on indicators displacement assay (IDA) using three metal ions (Cu²⁺, Zn²⁺ and Ni²⁺) as receptors to organic acids. The time stability results showed that the prepared CSA had good operational stability. Three quantitative models, including one linear (partial least square, PLS) and two nonlinear (support vector regression, SVR and back propagation artificial neural network, BP-ANN) models were used to estimate the content of total acids in fermentation broth of apple vinegar through image analysis. The correlation coefficient (RP), root mean square error of prediction (RMSEP) and residual predictive deviation (RPD) of the better SVR model were 0.8708, 0.0545 and 10.91, respectively. The results implied that the CSA had an excellent potential for quantitative monitoring of total acids in apple vinegar during fermentation.
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Background and Objectives The fetal period is an important stage in a person’s life. Exercise during pregnancy has been considered as a positive factor in preventing chronic diseases in adulthood. The aim of this study was to investigate the role of maternal high intensity exercise before and during pregnancy on serum levels of lipid profile of adult offspring. Subjects and Methods Twenty-four rats (170-200g) were divided into three maternal groups: Control (C), exercise Before Pregnancy (BP) and exercise before and during pregnancy (BDP). Exercise performed before pregnancy for six weeks and during pregnancy for three weeks; one minute with 80% -95% vo2max switching with two minutes of active recovery (65% vo2max) for five days/week. After end of the training birth process and breastfeeding, the offspring were divided according to theirs’s mother group and they were kept until adult age. The serum levels of LDL, HDL, TG and Cho were measured by enzymic method. Results The one-way ANOVA result showed that maternal exercise before and during pregnancy significantly reduced LDL, Cho and LDL/HDL cholesterol ratio in male (P=0.03, P=0.2, P=0.04) and female (P=0.00, P=0.02, P=0.04) offspring. Conclusion This finding suggests that maternal high-intensity-interval training as an appropriate environmental intervention can help to improve the health of the next generation.
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Since gastrointestinal mucosa is constantly exposed to reactive oxygen species from various sources, the presence of antioxidants may contribute to the body’s natural defenses against inflammatory diseases. Hypothesis To define the polyphenols extracted from dried apple peels (DAPP) and determine their antioxidant and anti-inflammatory potential in the intestine. Caco-2/15 cells were used to study the role of DAPP preventive actions against oxidative stress (OxS) and inflammation induced by iron-ascorbate (Fe/Asc) and lipopolysaccharide (LPS), respectively. Results The combination of HPLC with fluorescence detection, HPLC-ESI-MS TOF and UPLC-ESI-MS/MS QQQ allowed us to characterize the phenolic compounds present in the DAPP (phenolic acids, flavonol glycosides, flavan-3-ols, procyanidins). The addition of Fe/Asc to Caco-2/15 cells induced OxS as demonstrated by the rise in malondialdehyde, depletion of n-3 polyunsaturated fatty acids, and alterations in the activity of endogenous antioxidants (SOD, GPx, G-Red). However, preincubation with DAPP prevented Fe/Asc-mediated lipid peroxidation and counteracted LPS-mediated inflammation as evidenced by the down-regulation of cytokines (TNF-α and IL-6), and prostaglandin E2. The mechanisms of action triggered by DAPP induced also a down-regulation of cyclooxygenase-2 and nuclear factor-κB, respectively. These actions were accompanied by the induction of Nrf2 (orchestrating cellular antioxidant defenses and maintaining redox homeostasis), and PGC-1α (the “master controller” of mitochondrial biogenesis). Conclusion Our findings provide evidence of the capacity of DAPP to reduce OxS and inflammation, two pivotal processes involved in inflammatory bowel diseases.
This reference book contains a comprehensive selection of the most frequently used assays for reliably detecting pharmacological effects of potential drugs, including tests for cardiovascular, analgesic, psychotropic, metabolic, endocrine, respiratory, renal, and immunomodulatory activities. Each of the over 700 assays comprises a detailed protocol with the purpose and rationale of the method, a description of the experimental procedure, a critical assessment of the results and their pharmacological and clinical relevance, and pertinent references. Identification of specific tests is facilitated by the enclosed CD-ROM which allows for a quick and full text research. An appendix with guidelines and legal regulations for animal experiments in various countries will help to plan these experiments properly in accordance with the welfare of laboratory animals.