doi: 10.1111/fcp.

12144

ORIGINAL Ameliorative effect of statin therapy on

ARTICLE
oxidative damage in heart tissue of
Fundamental & Clinical Pharmacology

hypercholesterolemic rabbits
Volkan Sozer*
Department of Biochemistry, Yildiz Technical University, Istanbul, Turkey

Keywords
atorvastatin,
hypercholesterolemia,
malondialdehyde,
paraoxonase-1,
protein carbonyl
Received 4 March 2015;
revised 5 July 2015;
accepted 21 August 2015
*Correspondence and reprints:
vsozer@gmail.com
ABSTRACT
The aim of this study was to investigate the effects of a high-cholesterol diet in the
presence and absence of statin on Cu-Zn-superoxide dismutase (Cu,Zn-SOD),
malondialdehyde (MDA), protein carbonyl (PCO), and nitric oxide (NO) of blood
and heart tissue, the antioxidant activity of serum paraoxonase-1 (PON-1), and on
the blood lipid profile of rabbits. The animals were divided into four groups each of
which included 10 rabbits. Rabbits in group 1 received a regular rabbit chow diet
(normal diet) for 8 weeks; those in group 2 received atorvastatin (0.3 mg atorvas-
tatin per day/kg body weight) for 8 weeks; those in group 3 received high-choles-
terol diet for 8 weeks; and those in group 4 received high-cholesterol diet for
4 weeks, a high-cholesterol diet + atorvastatin (0.3 mg atorvastatin per day/kg
body weight) for 8 weeks. The parameters were measured by spectrophotometric
methods. As expected, the atherogenic diet caused a pronounced increase in lipid
profile (not HDL) parameters. Rabbits in group 3 showed higher PCO, MDA, and
NO levels in circulating and heart tissue compared to the rabbits in group 1. Ator-
vastatin has prevented or limited LDL oxidation and has showed constitutively
beneficial effects in group 4. Increased LDL-C, PCO, MDA, and NO levels leading to
decreasing PON-1 activity thus create a predisposition to atherogenesis in this
model. But atorvastatin administration partly ameliorated oxidative damage in
heart injury of hypercholesterolemic rabbits. Atorvastatin which functions as a
potent antioxidant agent may inhibit this LDL-C oxidation by increasing PON-1
activity in atherogenesis.

Protein oxidation is the covalent modification of a

INTRODUCTION
Hypercholesterolemia is the common risk factor of
coronary artery disease (CAD), and it is highly associ-
ated with obesity, diabetes mellitus, and several other
metabolic syndromes. Statins are potent inhibitors of
cholesterol biosynthesis, and they may be efficient on
reduction of morbidity and mortality of cardiovascular
diseases. There are some pleiotropic effects of statins
such as improvement of endothelial function, inflam-
mation, function of the platelets, and stabilization of
atherosclerotic plaque independent from reduction in
the cholesterol levels [1].
protein induced either directly by reactive oxygen
species (ROS) or indirectly by reaction with secondary
by-products of oxidative stress. Protein carbonylation is
the most salient change in oxidized proteins. The most
commonly measured products of protein oxidation in
biological samples are the protein carbonyls (PCOs) [2].
Malondialdehyde (MDA) is the end product of lipid per-
oxidation (LPO) and the biomarker of oxidative stress
in various cases. Nitric oxide (NO) is a unique gaseous
molecule capable of regulating various bodily func-
tions, either directly or via cGMP-mediated mechanism.
NO has a dual effect: when it is produced under

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Statin and oxidative damage
physiological concentration, it is beneficial; however,
excessive NO production under pathological conditions
can be destructive. Reduced bioavailability of NO is
thought to be one of the central factors common to
CAD, although it is unclear whether this is a cause or
a result of endothelial dysfunction [3].
Glutathione (GSH) is a sulfhydryl antioxidant; the
oxidized form is a sulfur–sulfur linked compound,
known as glutathione disulfide or GSSG. Paraoxonase
(PON) is an antioxidant enzyme closely associated with
high-density lipoproteins, which detoxifies lipid perox-
ides, and is widely distributed in many tissues. Oxidized
LDL phospholipids are physiological substrates for
PON-1 [4]. Superoxide dismutase (SOD) is an enzyme
that functions to catalytically convert O2• to O2 and
hydrogen peroxide. These are important antioxidant
defenses in nearly all cells exposed to oxygen. In
eukaryotic cells, two intracellular superoxide dismutase
exist: the Cu,Zn-SOD and the Mn-SOD. Cu,Zn-SOD is
the major intracellular SOD. These enzymes particu-
larly protect the red cells against O2• mediated LPO. In
addition to antioxidant effects, encapsulated SOD exhi-
bits lipid-lowering effects in fat-fed rats [5].
As the antioxidant activity of statins, particularly in
the heart, has not yet been well examined, we aimed
to investigate the role of atorvastatin in the heart of
rabbits. The action/impact of statin, a substance with
pleiotropic effect, on systemic circulation and heart has
been determined in a cholesterol-rich diet-induced
experimental atherogenesis model. PCO, MDA, NOx,
GSH, PON-1, and Cu,Zn-SOD activities have been
examined as a result of oxidative stress in rabbits.
MATERIALS AND METHODS
Animals and experimental procedures
Forty male rabbits (6-month-old New Zealand White)
weighing 2.5–3.4 kg and obtained from the center for
Experimental Animal-Research and Breeding Labora-
tory (Cerrahpasa Medical Faculty, Istanbul University,
Istanbul, Turkey) were used in this study. All rabbits
were housed in individual cages and were quarantined
for 7 days prior to the study and received standard pel-
lets (15% protein, 2.5% lipid, 15% cellulose, 14% clay,
13% water) during this time. The rabbits were housed
individually with 12-h light periods, at a temperature
of 20  2 °C. Food and fresh tap water were supplied
ad libitum throughout the experiment ‘The Guide for
the Care and Use of Laboratory Animals’. Interventions
concerning experimental animals were performed

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according to the US National Institutes of Health (NIH
Publication 85, 23, revised 1996) and under the 3 Rs
rule (Reduction, Replacement and Refinement).
Group 1 (normal diet, n = 10) were randomly cho-
sen to be fed with the normal diet. Group 2 (normal
diet+atorvastatin, n = 10) received standard pellets,
atorvastatin added until the end of week 8; a group of
rabbits (n = 20) were fed with normal rabbit chow diet
fortified with 1% cholesterol for 4 weeks. Hypercholes-
terolemic rabbits were randomly divided into two
groups, as group 3 (hypercholesterolemic diet, n = 10)
and group 4 (hypercholesterolemic diet+atorvastatin,
n = 10), for 8 weeks. Atorvastatin powder was sup-
plied from Sanovel, Istanbul. The dosage of atorvastatin
(0.3 mg atorvastatin per day/kg body weight) was
added to the diet of atorvastatin-treated rabbits under
study. Chow consumption of all the rabbits was moni-
tored and recorded daily.
Blood samples were drawn from the central ear
artery both at the beginning of the experimental period
and at 4 weeks later. At the end of the study period,
the animals were sacrificed using an overdose of pento-
barbital. Under anesthesia, the thorax was opened and
blood from the right ventricle was collected into dry
and heparinized tubes for determination of various
analytes. Heart tissue samples were obtained and kept
in a liquid nitrogen tank until analyses were per-
formed. The blood samples were centrifuged for 5 min
at 1000 9 g at 4 °C. The serum and plasma were
stored at 80 °C until analysis. The erythrocytes were
washed three times with an equal volume of cold sal-
ine (0.9%, v/v). One milliliter of washed-out erythro-
cytes was lysed with 3 mL deionized water (dH2O)
(1 : 3, v/v). All samples were extracted from lysates.
GSH was analyzed within 6 h after extraction.
Preparation of tissue samples
Heart tissues were removed and washed in saline.
About 180–200 mg of each sample was weighed and
diluted 20% w/v in 20 mM ice-cold Tris–HCl, pH = 7.4,
and homogenized with a Bosch Scintilla SA (Switzer-
land). The homogenate was centrifuged at 5000 9 g
for 10 min, and analyte determinations were per-
formed in the supernatant fraction.
Assay of PCO levels
Plasma and tissue PCO levels were measured spec-
trophotometrically using the method of Reznick and
Packer [6]. Protein content was determined on the HCl
blank pellets spectrophotometrically using a Folin kit
560
(Sigma Diagnostics, St. Louis, MO, USA). The coeffi-
cients of intra- and interassay variations for the PCO
assay were 4.0% (n = 15) and 7.2% (n = 15), respec-
tively.
Assay of MDA levels
Serum and tissue MDA levels were measured with the
colorimetric thiobarbituric acid method [7]. The coeffi-
cients of intra- and interassay variations for the MDA
assay were 3.7% (n = 15) and 5.2% (n = 15), respec-
tively.
Assay of NOx levels
Serum and tissue NOx levels were measured spec-
trophotometrically as its stable metabolites nitrate and
nitrite by the Griess reagent method with a commer-
cially available colorimetric assay (Boehringer Man-
nheim, GmBH, Germany). The intra-assay and
interassay coefficients of variation were 4.0% (n = 15)
and 5.2% (n = 15).
Assay of GSH levels
Erythrocyte and tissue GSH levels were measured spec-
trophotometrically using the method of Beutler et al.
[8]. The intra- and interassay coefficients of variation
for GSH were 3.2% (n = 15) and 4.3% (n = 15),
respectively.
Assay of PON-1 activity
Serum PON-1 activity was measured by spectropho-
tometry using synthetic paraoxon (diethyl-p-nitro-
phenyl phosphate) as substrate [9]. The intra-assay
and interassay coefficients of variation were 4.7%
(n = 15) and 5.3% (n = 15), respectively.
Assay of Cu,Zn-SOD activity
Serum and tissue Cu, Zn-SOD activity was measured
spectrophotometrically using the method of Sun
et al.[10]. The intra-assay and interassay coefficients of
variation were 3.5% (n = 15) and 4.0 (n = 15), respec-
tively.
Serum total protein, albumin, total cholesterol (TC),
high-density lipoprotein cholesterol (HDL-C), and
triglyceride (TG) levels were determined using kits from
Sigma (St Louis, MO, USA).
Data analysis
Statistical analyses were performed using SPSS 20.0
(SPSS Inc., Chicago, IL, USA) for Windows. For each
V. Sozer
variable, values were expressed as mean (SEM). Com-
parisons of parameters between the different groups
were made using the nonparametric Mann–Whitney
U-test. Statistical evaluations within the groups were
carried out using the Wilcoxon signed-rank test.
The relationships among the analyzed parameters
were investigated using Pearson’s correlation coeffi-
cients.
RESULTS
There were no significant differences in baseline serum
lipid concentrations among the groups. The hyperc-
holesterolemic diet caused a pronounced increase in
lipid parameters. Atorvastatin treatment reduced the
serum levels of TC, LDL-C, and TG compared to the
nontreated group (Table I).
There were no significant differences in initial PCO,
MDA, NOx, GSH, PON-1, and Cu,Zn-SOD activities
among all the groups (Table II). Supplementation of the
high-cholesterol diet for 4 weeks significantly increased
PCO, MDA, and NOx levels, whereas decreased activity
of GSH, PON-1, and Cu,Zn-SOD was observed in groups
3 and 4. Atorvastatin treatment caused an increase in
GSH, PON-1, and SOD activities and a decrease in
PCO, MDA, and NOx levels (Table II).
All oxidative stress parameters of the heart tissue
were not significantly different among groups 1 and 2
(Table III). PCO, MDA, and NOx levels (P < 0.001, for
all these parameters) in the heart tissues from the
high-cholesterol diet group were significantly higher
than the control groups (groups 1 and 2). Atorvastatin
therapy caused significant decreases in PCO, MDA, and
NOx levels (P < 0.001, P < 0.01, P < 0.05, respec-
tively). Tissue GSH of the high-cholesterol diet group
was significantly lower than the other groups
(P < 0.05). Atorvastatin treatment caused an increase
in GSH levels in group 4, but this increase was not sta-
tistically significant in comparison with other groups.
Cu, Zn-SOD activities of the tissue was not significantly
different among the studied groups.
The PON-1 activity was negatively correlated with
the TC, LDL-C, TC/HDL-C, and LDL-C/HDL-C levels in
the cholesterol-fed rabbits (r = 0.856, P = 0.002;
r = 0.859, P = P = 0.001; r = 0.766, P = 0.010;
r = 0.781, P = 0.008, respectively) (Figure 1). Pear-
son’s correlation coefficients between PCO and GSH
levels in heart tissue of the cholesterol-fed rabbits
(r = 0.843, P = 0.002) are shown in Figure 2.

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Statin and oxidative damage 561

Table I Effects on lipid levels of hypercholesterolemic and atorvastatin diet throughout the experiment.

Group 1 (n = 10) Group 2 (n = 10) Group 3 (n = 10) Group 4 (n = 10)

TC (mg/dL)
Baseline 100.10  14.68 95.60  13.41 100.40  16.97 96.70  16.19
Week 4 98.50  15.14 91.60  10.15 362.20  56.27***,a,b,x,z 386.10  35.39***,a,b,x,z
Week 8 103.00  18.32 87.70  11.96 349.50  58.7***,a,b,x,y 288.90  47.56***,a,b,c,x,y
LDL-C (mg/dL)
Baseline 56.76  14.95 54.98  14.23 58.42  18.40 54.68  15.63
Week 4 53.60  18.40 49.86  9.33 319.82  55.20***,a,b,x 339.32  35.40***,a,b,x
Week 8 61.74  15.77 47.38  10.41 310.10  59.63***,a,b,d,x 244.42  44.48***,a,b,x
HDL-C (mg/dL)
Baseline 24.50  4.88 23.00  4.24 23.30  4.55 24.20  5.22
Week 4 24.80  4.85 22.40  2.91 23.40  4.81 27.30  4.42*,a,b,x,z
Week 8 24.20  5.81 23.40  4.81 20.20  2.25***,a,b,c,x,*,y 31.80  4.16***,a,b,c,x,*,y
Triglycerides (mg/dL)
Baseline 94.20  18.20 88.10  17.82 93.40  19.55 89.10  15.85
Week 4 100.50  25.87 96.70  20.61 94.90  8.69 97.40  10.80*,x,***,z
Week 8 85.30  28.81 84.70  28.15 96.00  20.34*,a,b,***,d 63.40  7.38***,a,b,c,x,y
TC/HDL
Baseline 4.25  1.11 4.25  0.80 4.51  1.38 4.13  0.91
Week 4 4.21  1.51 4.14  0.64 16.01  3.67***,a,b,x 14.59  3.42***,a,b,x,z
Week 8 4.50  1.60 3.85  0.79 17.38  2.73 ***,a,b,d,x 9.12  1.14***,a,b,c,x,y
LDL/HDL
Baseline 2.45  0.92 2.46  0.77 2.65  1.17 2.34  0.72
Week4 2.37  1.31 2.26  0.50 14.17  3.54***,a,b,x 12.86  3.29***,a,b,x,z
Week 8 2.74  1.21 2.11  0.71 15.42  2.77***,a,b,x 7.71  1.10***,a,b,x,y

Group 1, normal diet; group 2, normal diet+atorvastatin; group 3, hypercholesterolemic diet; group 4, hypercholesterolemic diet + atorvastatin; TC, total
cholesterol; HDL-C, high-density lipoprotein; LDL-C, low density lipoprotein.
* P < 0.05, *** P < 0.001.
a
vs group 1; bvs group 2; cvs group 3; and dvs group 4.
x
vs baseline; yvs week 4; and zvs week 8 within the same group.

PON-1, and SOD are measured. The PCO measure reac-

DISCUSSION
The present study showed that PCO, MDA, and NOx
levels were higher in hypercholesterolemic diet as com-
pared to normal diet. As it was expected, the hyperc-
holesterolemic diet group had higher TC and LDL levels
than the normal-diet group. There were positive corre-
lations between LDL-C and PON-1 activities only in
cholesterol-rich diet-induced rabbits. The relationship
between LDL-C and PON-1 can be explained through
the effect of PON-1 on atherogenesis. PON-1 plays a
central role in determining LDL-C clearance from the
blood via hydrolyzes oxidized lipid or lipid hydroperox-
ides in LDL.
Consistent with our previous work as well as with
other reports [11–15], the present study demonstrated
that the coexistence of severe hypercholesterolemia
enhanced oxidative stress and atherogenesis in the
cholesterol-rich diet-induced model. As an index of
antioxidative defense system, PCO, MDA, NOx, GSH,
tive aldehydes formed as end products of oxidative
reactions. The increased aldehyde levels are evidently
shown by their considerable role in the pathogenesis of
atherogenesis. In the present study, plasma and heart
tissue PCO levels are increased in the cholesterol-fed
rabbits. The PCO levels were also negatively correlated
with the GSH levels in heart tissue. The oxidative mod-
ification of proteins is generally used as a marker of
oxidative damage, which is increased with the patho-
physiology of atherogenesis, and has even been sug-
gested as a causative factor of the progress of
atherogenesis. In the present study, atorvastatin ther-
apy, which is a potent inhibitor of cholesterol biosyn-
thesis, caused significant decreases in both plasma and
tissue PCO levels in cholesterol-rich diet-induced rab-
bits.
MDA, a convenient biomarker of LPO, was therefore
used to evaluate the robust effect of atorvastatin on
LPO in circulating and heart tissue. Atorvastatin

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562 V. Sozer

Table II Effects on oxidative stress parameters of hypercholesterolemic and atorvastatin diet throughout the experiment.

Group 1 (n = 10) Group 2 (n = 10) Group 3 (n = 10) Group 4 (n = 10)

Plasma PCO (mg/dL)

Baseline 0.76  0.13 0.67  0.15 0.70  0.16 0.72  0.13
Week 4 0.78  0.14 0.64  0.18 1.16  0.19***,a,b,x,**,z 1.04  0.15***,a,b,x,**,z
Week 8 0.77  0.17 0.65  0.18 1.37  0.15***,a,b,x,**,y 0.88  0.23***,a,b,c,x,**,y
Serum MDA (mg/dL)
Baseline 2.01  0.34 2.35  0.60 2.18  0.26 2.33  0.38
Week 4 2.26  0.42 2.09  0.50 4.16  0.30***,a,b,x,*,z 3.30  0.52***,a,b,x,**,z
Week 8 2.20  0.64 2.23  0.63 4.47  0.62***,a,b,x,*,y 2.82  0.28**,a,b,x,y,***,c
Serum NOx (mg/dL)
Baseline 38.00  6.70 43.50  8.83 42.90  7.77 42.50  6.55
Week 4 43.90  7.92 40.50  5.08 82.20  14.08***,a,b,x 79.40  11.65***,a,b,x,z
Week 8 43.10  7.16 44.10  10.43 82.20  22.17*** ,a,b,x
59.40  13.43**,a,b,x,***,c,y
Erythrocyte GSH (mg/dL)
Baseline 4.69  0.37 4.65  0.39 4.89  0.22 4.83  0.32
Week 4 4.75  0.31 4.94  0.35 3.29  0.31***,a,b,x,*,z 3.10  0.48***,a,b,x,**,z
Week 8 4.58  0.62 4.87  0.66 2.86  0.40***,a,b,x,*,y 3.59  0.60**,a,b,c,x,y
Serum PON-1 (mg/dL)
Baseline 127.20  24.62 125.90  21.31 125.80  16.19 122.70  15.54
Week 4 120.80  6.77 145.80  8.68 81.60  10.89***,a,b,x,z 90.10  10.31***,a,b,x,*,z
Week 8 126.30  19.66 143.90  20.29 52.90  7.09***,a,b,d,x,y 96.40  17.33***,a,b,c,x,*,y
Serum CuZn-SOD (mg/dL)
Baseline 25.40  3.39 23.67  3.50 26.78  3.22 25.15  2.88
Week 4 24.09  3.66 24.29  2.27 21.86  2.76*,a,b,,z,**,x 21.51  2.04*,a,b,x
Week 8 25.22  2.04 24.43  3.13 19.24  2.28 ***,a,b,,x,**,d,*,,y 23.13  2.88**,c,*,y

SOD, superoxide dismutase; NOx, total nitric oxide; PCO, protein carbonyl; GSH, glutathione; Hb, hemoglobin.
* P < 0.05, ** P < 0.01, *** P < 0.001.
a
vs group 1; bvs group 2; cvs group 3; and dvs group 4.
x
vs baseline; yvs week 4; and zvs week 8 within the same group.

Table III Effects of atorvastatin supplementation (12 weeks) on heart tissue of rabbits fed with hypercholesterolemic diet.

Group 1 (n = 10) Group 2 (n = 10) Group 3 (n = 10) Group 4 (n = 10)

GSH (lmol/g pr.) 77.65  12.92 73.88  16.19 59.88  14.04*,a,b 68.58  14.88
SOD (U/g pr.) 30.45  4.59 31.62  4.84 30.12  7.15 31.19  4.71
PCO (nmol/mg pr.) 1.66  0.27 1.43  0.25 2.49  0.36***,a,b,d 1.80  0.43***,c
MDA (nmol/g pr.) 2.08  0.41 1.99  0.52 3.28  0.53*** ,a,b,d
2.37  0.36**,c
NOx (lmol/g pr.) 13.88  2.39 13.85  2.29 21.94  2.98***,a,b,*,d 17.75  3.73*,a,b,c

* P < 0.05, ** P < 0.01, *** P < 0.001.

a
vs group 1, bvs group 2, cvs group 3, dvs group 4.

treatment caused a decrease in MDA, whereas the
high-cholesterol diet increased MDA of blood and heart
tissue in cholesterol-rich diet-induced rabbits. MDA, as
an indicator of LPO and biomarker of oxidative stress,
increases in cholesterol-rich-diet. Hypercholesterolemia
might be attributed to LPO and excessive MDA
production has been associated with atherogenesis.
Increased MDA seems to play a key role in the onset of
atherogenesis, which is considered one of the main
processes leading to CAD. The possible antihypercholes-
terolemia mechanism of atorvastatin may regulate the
disorder of lipid metabolism as well as enhance cardiac
function. These results were consistent with previous
findings [13–20] and further supported the concept of
the pleiotropic vascular protection effects of atorvas-
tatin in hypercholesterolemia.

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Statin and oxidative damage
(a)
(c)
Figure 1 The relationship between PON-1 with TC (a), LDL-C (b), TC/HDL-C (c), and LDL-C/HDL-C (d) in the cholesterol-fed rabbits.
Figure 2 The relationship between PCO and GSH levels in heart
tissue of the cholesterol-fed rabbits.
In the presence of atherogenesis or risk factors such
as hypercholesterolemia, NO levels are decreased [21].
However, the results regarding NO levels increasing/de-
creasing effect of statins are conflicting. In the present
study, serum and tissue NO of the cholesterol-rich-diet

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(b)
(d)
group was higher than normal-diet groups, while NO
was decreased in group 4. Hypercholesterolemic ani-
mals had higher serum NO compared with normal-diet
animals, which supports the results of previous studies
[22–26]. The novelty of the present study lies in the
fact that increased NO production in circulating and
heart tissue worsens atherogenic conditions. But, ator-
vastatin may be a protective response of endothelial
cells in the early stage of atherogenesis and these
effects were independent of lipoprotein changes. Serum
NO levels have been also reported to be lower in vari-
ous studies [13,21,27,28]. Bolayirli et al. [13] reported
that NO levels significantly decreased in hypercholes-
terolemic rabbits, whereas atorvastatin therapy did not
result in a significant change in NO in hypercholes-
terolemic rabbits. Increased NO by modulating eNOS
may be another possible mechanism of the vasoprotec-
tive effects of statins. Further studies are needed to
show the effects of statins for damage induced by
hypercholesterolemia.
564
GSH can produce coronary vasodilation when added
to isolated, perfused rodent heart, very likely due to its
normalizing effect on prostaglandin synthesis. Results
of the present study have indicated that the erythro-
cyte GSH levels decreased in cholesterol-fed animals
compared with normal diet and hypercholesterolemic
diet + atorvastatin. This study showed that the athero-
genic diet caused a significant decrease in GSH levels of
erythrocyte and heart tissue. Atorvastatin therapy
caused significant increase in GSH levels in cholesterol-
rich diet-induced rabbits. Atorvastatin therapy has sub-
stantially beneficial effects on the oxidant stress
induced by hypercholesterolemia. The effect of statins
on GSH levels is paradoxical [11,29–32]. Save et al.
[29] reported that plasma levels of GSH were increased
by 10 mg/day atorvastatin in hypercholesterolemic
patients. Our other study [11] demonstrated that ator-
vastatin therapy caused significant decreases in both
PCO and MDA levels, but a significant increase in ery-
throcyte GSH levels. In the study by Liu et al. [30],
10 mg/day atorvastatin did not affect the erythrocyte
GSH level. In a comparative dose-dependent study of
atorvastatin, simvastatin, and pravastatin, erythrocyte
GSH remained unchanged after statin treatment in
hypercholesterolemic patients [31]. The GSH system (in
erythrocyte and heart tissue), an important protective
system against oxidative damage, may be affected by
statins. However, different types of statins may expend
different effects on GSH levels. The GSH, an important
protective system against oxidative damage in hyperc-
holesterolemic state, may be prevented or modulated
by atorvastatin. GSH as a potent antioxidant may be a
therapeutic option. Future trials are indicated with sta-
tins, administrated in combination with other antioxi-
dants such as GSH in hypercholesterolemia.
Supplementation of the high-cholesterol diet for
4 weeks decreased in serum PON-1 activity, whereas
4 weeks after atorvastatin treatment, the PON-1 activity
increased in the present study. The decreased PON-1
may be caused by the increase in oxidative stress
induced by high-cholesterol. Similar to other studies’
results, this study has shown that PON-1 is another
antioxidant enzyme like platelet activating factor acetyl-
hydrolase associated with HDL, which detoxifies LPO.
Harangi et al. [33] has demonstrated that atorvastatin
treatment favorably affected the lipid profile, increasing
the HDL-associated PON activity in type-II/a hyperlipi-
demic patients. In another study, atorvastatin therapy in
dyslipidemic patients decreases the level of oxidative
stress and increases PON activity, especially in patients
V. Sozer
with HDL levels above 35 mg/dL [34]. Paragh et al.
[35] have shown that short-term administration of sim-
vastatin did not increase PON activity and atorvastatin
treatment had a favorable effect on lipid profile and
increased the activity of HDL-associated PON. PON,
which prevents LDL oxidation and inactivates LDL-
derived oxidized phospholipids, showed a pronounced
decrease in the group receiving the atherogenic diet, and
atorvastatin increased PON activity [18]. In the present
study, the changes in PON-1 activity overlap with the
results of the study reported by other authors
[13,18,33–37]. These profitable effects may be attributed
to the antioxidant properties of statins and the increase
in PON-1 activity in cholesterol-rich diet-induced rabbits.
Oxidative stress plays a critical role in the develop-
ment of atherogenesis. In addition, decreased activities
of SOD, catalase (CAT), and glutathione peroxidase
(GPx) may lead to the accelerated progression of
atherogenesis. There has been substantial interest in
the level of expression of various antioxidant defense
mechanisms, and in particular that of SOD in
atherosclerotic vessels [38]. SOD activity has been
reported to be unaffected and higher in previous stud-
ies with atherogenesis [38–43]. In vessels of apo E–defi-
cient mice, extracellular SOD activity and protein
expression are increased by two- to threefold, whereas
the activities of the cytosolic CuZnSOD and MnSOD are
not changed. Codo~ ner-Franch et al. [44] found that
SOD activity was reduced in hypercholesterolemic chil-
dren compared to the control group. Hypercholes-
terolemia decreased serum CuZn-SOD, but not in heart
tissue in the present study. Atorvastatin did not have
any effect on heart tissue, while serum CuZn-SOD
activity was increased in the rabbits treated with ator-
vastatin in comparison with high-cholesterol diet
group. Sezer et al. [18] show that an atherogenic diet
caused a prominent increase in SOD activity. Increased
SOD activity represents an important physiological
adaptation counteracting the increase in oxidative
stress. Decreased SOD activity in erythrocytes, after
8 weeks of atorvastatin treatment, indicates the antiox-
idant effect of the drug [27,45–48]. Statins have been
widely used in clinical practice owing to its both potent
lipid-modifying effects and other cardio-protective
effects including increasing/decreasing SOD generation.
CONCLUSION
The data indicated that hypercholesterolemia
contributes to atherogenesis progression and that

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Fundamental & Clinical Pharmacology 29 (2015) 558–566
Statin and oxidative damage
atherogenesis induces oxidative damage in circulating
and heart tissue. Atorvastatin attenuates the hyperc-
holesterolemic atherogenesis in the rabbit model.
Because of the all aforementioned benefits regarding
atorvastatin therapy, that is decreased oxidative stress
in the heart and improved the lipid profile in choles-
terol-fed rabbits, it may be considered a useful tool for
the reduction of oxidative stress and improvement of
lipid profiles in diseases related to atherosclerosis. Fur-
ther studies are needed to understand the complete
mechanism and potential actions of statins.
ACKNOWLEDGEMENTS
In memoriam: This article is dedicated to the memory
of Professor Tuncay Altug, Experimental Animal-Re-
search and Breeding Laboratory, Istanbul University,
Cerrahpasa Faculty of Medicine, who kindly provided
experimental animals for my current research. I want
to thank Prof. Dr. Hafize Uzun for giving me the
chance to study in her laboratory and supporting me
along the study period.
DECLARATION OF INTEREST
There was no pharmaceutical company involvement
with this article.
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