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Pharmacological Reports
Copyright © 2012
2012, 64, 643–649
by Institute of Pharmacology
ISSN 1734-1140
Polish Academy of Sciences
Granulocyte colony-stimulating factor improves
early remodeling in isoproterenol-induced cardiac
injury in rats
Ludimila Forechi, Marcelo P. Baldo, Diana Meyerfreund, José G. Mill
Department of Physiological Sciences, Federal University of Espirito Santo, Av. Marechal Campos 1468, Maruipe,
29042-755, Vitória, Espírito Santo, Brazil
Correspondence: José G. Mill, e-mail: [email protected]; [email protected]
Abstract:
Background: Granulocyte colony-stimulating factor (G-CSF) has been used in some animal models and humans with wellestablished cardiovascular diseases. However, its effects in the initial stage of progressive non-ischemic heart failure are unknown.
Methods: Wistar rats (260–300 g) were divided into three groups: control (without any intervention), ISO (150 mg/kg isoproterenol
hydrochloride sc, once a day for two consecutive days), and ISO-GCSF (50 µg/kg/d G-CSF for 7 days beginning 24 h after the last
administration of ISO). Echocardiography was performed at baseline and after 30 days of follow-up. Subsequently, animals were
anesthetized for hemodynamic analysis. The left ventricle was removed for analysis of interstitial collagen deposition and cardiomyocyte hypertrophy.
Results: Isoproterenol led to left ventricular dilation (control, 7.7 ± 0.14 mm; ISO, 8.7 ± 0.16 mm; ISO-GCSF 7.8 ± 0.09 mm; p <
0.05), myocardial fibrosis (control, 2.0 ± 0.18%; ISO, 9.1 ± 0.81%; ISO-GCSF 5.9 ± 0.58%; p < 0.05) and cardiomyocyte hypertrophy (control, 303 ± 10 µm2; ISO, 356 ± 18 µm2; ISO-GCSF 338 ± 11 µm2; p < 0.05). However, G-CSF partially prevented collagen
deposition and left ventricular enlargement, with a slight effect on hypertrophy. Characterizing a compensated stage of disease, hemodynamic analysis did not change.
Conclusion: G-CSF administered for 7 days was effective in preventing the onset of ventricular remodeling induced by high-dose
isoproterenol with decreased collagen deposition and chamber preservation.
Key words: isoproterenol, heart failure, G-CSF, collagen
Abbreviations: CVF – collagen volume fraction, dP/dt –
maximal rate of pressure, FS – fractional shortening, G-CSF –
granulocyte colony-stimulating factor, HR – heart rate, ISO –
isoproterenol, LVEDD – left ventricular end-diastolic dimension, LVEDP – left ventricle end-diastolic pressure, LVESD –
left ventricular peak systolic dimension, LVSP – left ventricle
systolic pressure, MCSA – myocyte cross sectional area
Introduction
Massive sympathetic discharge, as observed in extreme stress or with high activity of the central nervous system, has deleterious effects on the heart. In
these cases, high oxygen consumption by the myocardium leads to subendocardial ischemia and infarction
followed by high levels of oxidative stress, ventricular dilation, fibrosis, progressive systolic dysfunction
and heart failure [6, 17, 29].
Non-invasive animal models have been used to
mimic the pathophysiological characteristics of this
cardiomyopathy in humans [7]. Rats treated with high
doses of the nonselective b-adrenergic receptor agonist isoproterenol are a widely used model to assess
progressive left ventricular dysfunction. Although the
mechanisms by which isoproterenol induces myocardial injury are not completely understood, b-adrenergic receptor overstimulation leads to intense coroPharmacological Reports, 2012, 64,
643–649
643
nary vasoconstriction and the development of microinfarcts that more frequently affect the subendocardium or transmural region of the left ventricle wall
[10, 27, 28]. Teerlink et al. [29] showed progressive
left ventricular dilation and dysfunction associated
with long-term mortality rates after high doses of isoproterenol.
Recently, the therapeutic use of granulocyte
colony-stimulating factor (G-CSF) has been reported
in some tissues, such as heart, kidney and brain [4, 21,
31]. Studies have suggested that this cytokine can act
in damaged hearts by recruiting bone marrow-derived
cells and promoting angiogenesis, reducing apoptosis
and modulating collagen content, thus improving
overall cardiac function [1, 2, 12, 16, 19, 22, 23].
Most of these studies have been performed in different models of ischemic cardiomyopathy. Little is
known about the potential cardioprotective effects of
G-CSF in non-ischemic cardiomyopathies. Therefore,
this study sought to evaluate the effects of G-CSF
treatment on structural and functional cardiac parameters during cardiomyopathy induced by high-dose isoproterenol in rats.
Echocardiography
Transthoracic echocardiography was performed at
baseline and 4 weeks after isoproterenol treatment as
previously described [9]. Rats were intraperitoneally
anesthetized with ketamine (50 mg/kg; Agener, Brazil) plus xylazine (10 mg/kg; Bayer, Brazil) and
placed in the supine position, and electrodes were
placed in the subcutaneous tissue of limbs for continuous electrocardiogram recordings. M-mode images
were acquired with an echocardiograph (Aplio XG
SSA-790, Toshiba, Japan) using a 7.5 MHz transducer
transverse to the left ventricle at the level of papillary
muscles. M-mode values of left ventricular enddiastolic dimension (LVEDD) and left ventricular
peak systolic dimension (LVESD) were measured for
each animal as the average of four consecutive cardiac cycles under regular rhythm. Heart rate (HR),
fractional shortening {FS = ((LVEDD – LVESD)/
LVEDD) × 100)} and ejection fraction were obtained.
The measures were performed by an investigator
blinded to groups at both times.
Hemodynamic analysis
Materials and Methods
Animals and groups
Male Wistar rats (260–300 g) were acquired from the
animal facility at the Federal University of Espírito
Santo and randomly assigned into three groups: a control group, without any intervention; the ISO group,
which received isoproterenol hydrochloride (150 mg/kg,
sc, Sigma-Aldrich Inc., USA) once a day for two consecutive days; and the ISO-GCSF group, which received G-CSF (Biosintetica, 50 µg/kg, sc) treatment
for 7 days beginning 24 h after the last isoproterenol
dose. Cardiac function in these animals was studied
3 weeks after the last G-CSF dose. Animals had free
access to water and standard rat chow throughout the
experimental period, and all of the procedures were
performed in accordance with the Guide for the Care
and Use of Laboratory Animals (NIH Publication No.
85–23, revised 1996) and were approved by the institutional animal research committee (No. 012/2010).
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Four weeks after the first isoproterenol dose, animals
were anesthetized with ketamine (90 mg/kg) and xylazine (10 mg/kg), and hemodynamic measurements
were performed as previously reported [3]. Briefly,
the right common carotid artery was dissected to insert a fluid-filled polyethylene catheter (P50) connected to a blood pressure transducer (TRI 21, Letica
Scientific Instruments, Spain). The catheter was then
advanced into the left ventricle to record peak systolic
pressure (LVSP), end-diastolic pressure (LVEDP),
and heart rate (HR) in cardiac cycles under regular
cardiac beats. The maximal rate of pressure rise and
fall (+dP/dt and –dP/dt) was obtained electronically
(Powerlab/4SP ML750, AdInstruments, Australia)
and used as left ventricular contractility and relaxation indexes, respectively.
Determination of myocyte hypertrophy and
fibrosis
After hemodynamic recordings, the heart was removed and rapidly washed with cold saline solution,
and the ventricles were separated from the atria, blotted dry and weighed. The left ventricle was divided
into three slices of approximately 2 mm, which were
G-CSF prevents early cardiomyopathy in rats
Ludimila Forechi et al.
prepared for histology. Each slice was serially cut into
6 µm-thick transverse sections and stained with Sirius
red to determine collagen volume fraction (CVF).
Slices were also stained with hematoxylin-eosin to
determine the myocyte cross sectional area (MCSA).
The percentage of picrosirius red staining, which
indicated CVF, was measured in images obtained with
a video camera (Olympus, X-750, Indonesia) coupled
to an optical microscope (Top Light B2, Bel Engineering, Italy) under 400× magnification. Nine areas
of high-power fields were analyzed in the subendocardial layer, and nine were analyzed in the subepicardial layer. For MCSA evaluation, 40 to 60 myocytes
positioned perpendicularly to the plane of the section
with a visible nucleus and a clearly outlined and unbroken cell membrane were selected in each animal.
Cell images viewed with a video camera were projected onto a monitor and traced. Images for CVF and
MCSA evaluation were processed with Image J software (v. 1.43u, National Institutes of Health, USA).
Statistical analysis
All of the data are expressed as the mean ± standard
error of the mean (SEM). The goodness of fit for normal distribution was evaluated using the Kolmogorov-Smirnov test. Levene’s test was used to assess
the equality of variances. One-way analysis of variance (ANOVA) was used to compare means between
the experimental groups. Comparison among repeated
measurements of echocardiography was assessed by
a two-way ANOVA, followed by Fischer’s post-hoc
test for multiples comparisons. Statistical significance
was set at p < 0.05.
Results
General characteristics
The mortality during the 48 h of isoproterenol treatment was 18%, with the majority (67%) occurring after the first dose. Soon after each isoproterenol administration, signs of agitation and pelage erection
were evident in treated animals. While body weight
gain was observed in control rats, body weight loss
was evident in both groups treated with isoproterenol
(control: 12 ± 2.5 g; ISO: –12 ± 2.4 g; ISO + G-CSF:
–17 ± 1.7 g; p < 0.05). However, the acute body
weight loss observed in isoproterenol-treated rats was
fully recovered during follow-up. As observed in
Table 1, no significant difference was observed in relation to the morphometric parameters at the end of
the follow-up period.
Functional and structural assessment
Comparing the evolution of the echocardiographic parameters, an increased end-diastolic dimension was
Tab. 1. Morphometric and hemodynamic parameters four weeks after isoproterenol treatment
Sample size (n)
BW (g)
LV/BW (mg/g)
RV/BW (mg/g)
HR (bpm)
SBP (mmHg)
DBP (mmHg)
LVSP (mmHg)
LVEDP (mmHg)
dP/dt+ (mmHg/s)
dP/dt– (mmHg/s)
Control
ISO
ISO-GCSF
12
401 ± 9
1.99 ± 0.04
0.51 ± 0.01
245 ± 6
114 ± 4.1
84 ± 4.1
127 ± 5
7 ± 0.7
4856 ± 188
3369 ± 196
19
411 ± 7
2.01 ± 0.03
0.49 ± 0.01
241 ± 6
113 ± 2.9
82 ± 2.9
126 ± 3
7 ± 0.6
4806 ± 133
3731 ± 173
16
396 ± 6
2.03 ± 0.04
0.50 ± 0.01
235 ± 6
113 ± 2.9
83 ± 2.4
126 ± 4
7 ± 0.9
4775 ± 181
3593 ± 177
BW – body weight; LV – left ventricle; RV – right ventricle; HR – heart rate; SBP – systolic blood pressure; DBP – diastolic blood pressure; LVSP –
left ventricular systolic pressure; LVEDP – left ventricular end diastolic pressure; dP/dt – maximum rate of pressure rise and fall. Data are depicted as the mean ± SEM
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Tab. 2. Echocardiographic parameters at baseline and after four weeks of follow-up
LVEDD (mm)
Baseline
4 weeks
Variation
LVESD (mm)
Baseline
4 weeks
Variation
EF (%)
Baseline
4 weeks
Variation
FS (%)
Baseline
4 weeks
Variation
Control
(n = 8)
ISO
(n = 8)
ISO-GCSF
(n = 7)
7.0 ± 0.24
7.7 ± 0.14
0.7 ± 0.25
7.1 ± 0.12
8.7 ± 0.16*
1.6 ± 0.12*
7.2 ± 0.11
7.8 ± 0.09#
0.6 ± 0.15#
3.5 ± 0.16
3.9 ± 0.21
0.4 ± 0.21
3.8 ± 0.13
4.8 ± 0.22*
1.0 ± 0.22*
3.9 ± 0.15
4.1 ± 0.10#
0.2 ± 0.09#
82.5 ± 2.55
84.9 ± 1.93
2.4 ± 1.53
81.8 ± 1.23
80.9 ± 1.82
-0.8 ± 2.51
79.0 ± 3.05
83.5 ± 1.55
4.6 ± 3.04
49.8 ± 1.00
49.2 ± 2.06
–0.6 ± 2.22
47.4 ± 1.18
45.6 ± 1.63
–1.8 ± 2.15
46.0 ± 1.58
47.9 ± 1.26
1.9 ± 0.85
LVEDD – left ventricular end diastolic dimensions; LVESD – left ventricular systolic dimensions; EF – ejection fraction; FS – shortening fraction.
* p < 0.05 vs. CONTROL; # p < 0.05 vs. ISO
observed in all three groups (Tab. 2). In animals
treated with isoproterenol, the increase was higher
compared with other groups. The left ventricular
end-systolic dimension was also increased in these
animals. Treatment with G-CSF was effective in preventing left ventricular enlargement, as evaluated by
the end-diastolic diameter. Fractional shortening and
ejection fraction did not differ significantly between
groups. Moreover, hemodynamic analysis did not
change in any parameter evaluated (Tab. 1).
Morphological evaluation
When myocardial fibrosis was assessed in Sirius redstained sections, we found higher myocardium collagen content in rats receiving isoproterenol than in the
control group. G-CSF treatment attenuated the increase
in the collagen fractional volume induced by isoproterenol (control: 2.0 ± 0.18%; ISO: 9.1 ± 0.81%;
ISO-GCSF: 5.9 ± 0.58%; p < 0.05). The transverse
area of cardiomyocytes was higher after isoproterenol
administration, which was partially reversed by
G-CSF (control: 303 ± 10 µm2; ISO: 356 ± 18 µm2;
ISO-GCSF: 338 ± 11 µm2; p < 0.05) (Fig. 1).
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Discussion
Our study showed that a high isoproterenol dose administered during two consecutive days induces ventricular remodeling and increases collagen in the left
ventricle wall in rats. These changes can be partially
prevented by G-CSF because animals treated with this
cytokine 7 days after isoproterenol administration
clearly showed less ventricular dilation and reduced
collagen content in the left ventricular myocardium.
Teerlink et al. [29] described the kinetics of cardiac
structural and functional changes after different doses
of isoproterenol in rats. They showed progressive left
ventricular dilation at all tested doses (85, 170, and
340 mg/kg) up to 16 weeks of follow-up through passive pressure-volume curves but without affecting
hemodynamic parameters. Similar results were found
in our study, in which isoproterenol-treated rats
(150 mg/kg) showed similar results regarding left
ventricular dilation, as evaluated by echocardiographic analysis, but without hemodynamic repercussions. These data are in accordance with Teerlink et
al. [29] because the animals presented an initial stage
of cardiomyopathy development. Grimm et al. [11], in
G-CSF prevents early cardiomyopathy in rats
Ludimila Forechi et al.
Fig. 1. Histological analysis of the left ventricle 4 weeks after isoproterenol. (A) Representative images of hematoxylin-eosin- (top) and picrosirius red-stained (below) sections. (B) Myocyte cross-sectional area and (C) collagen volume fraction. Images were acquired at 400´ magnifi-
cation. * p < 0.05 vs. control; # p < 0.05 vs. ISO
contrast, reported increased left ventricular enddiastolic pressure only after two weeks of a single
dose of isoproterenol (150 mg/kg). These data, however, were not confirmed in our study.
Several studies have shown that G-CSF improves
cardiac function in animals submitted to ischemic injury by preventing enlargement of cardiac cavities
and reducing mortality [14, 16, 20, 25]. The mecha-
nism by which G-CSF acts on the cardiovascular system remains under intense discussion. Several studies
have shown that these beneficial effects may depend
on the interaction of different mechanisms, such as interference in anti-apoptotic signaling [1, 12], myocyte
regeneration [8], and modulation of synthesis and
degradation of components in the extracellular matrix
[19]. In this paper, we observed that G-CSF prevents
Pharmacological Reports, 2012, 64,
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early dilation, as evaluated by left ventricular diastolic dimension four weeks after a high dose of isoproterenol. These results suggest that acute G-CSF
can prevent the early structural changes leading to
systolic dysfunction and heart failure development after catecholaminergic aggression. It was previously
described that adrenergic overstimulation leads to
myocyte apoptosis [26]. Moreover, apoptosis is directly involved in heart failure progression [24]. Studies have shown that G-CSF therapy seems to reduce
heart failure progression through an antiapoptotic effect [1, 12, 13]. For example, G-CSF reduced FasL
expression and cardiomyocyte apoptosis in rats with
heart failure [13]. Thus, the reduction in left ventricular dilation observed in our work could be explained
by a reduction in cardiomyocyte apoptosis by early
G-CSF treatment.
Cardiac hypertrophy is another effect associated
with high-dose isoproterenol administration and the
first step to cardiac decompensation. It was reported
that high isoproterenol doses (85 and 150 mg/kg) led
to left ventricular hypertrophy [5, 11]. We observed
slight myocyte hypertrophy caused by isoproterenol,
despite no significant changes in the left ventricular
mass. It is likely that the increased volume of remaining myocytes compensated for the myocyte loss,
mainly in the subendocardial layer, in which fibrosis
is more intense. Similarly, Teerlink et al. [29] did not
observe differences in left ventricular weight early
after isoproterenol administration. G-CSF reduced
myocyte hypertrophy, which can be explained by
reduction of left ventricular dilation in G-CSF-treated
animals.
In the present study, isoproterenol increased collagen deposition in the myocardium. This finding is in
accordance with a previous description in which
a high dose of isoproterenol induced intense collagen
deposition [5]. G-CSF partially prevented collagen
deposition. Several studies have also shown that GCSF decreases myocardial fibrosis after cardiac damage [15, 19, 22]. This G-CSF effect does not seem to
be restricted to hypoxic damage because Macambira
et al. [17] recently observed that repeated injections
of G-CSF decreased inflammation and fibrosis in
a mouse model of Chagasic cardiomyopathy.
We cannot rule out the possibility that the absence
of hemodynamic impairment could be due the cardiodepressor effect of ketamine plus xylazine anesthesia.
However, similar results were found by Teerlink et al.
[29] using the same pharmacological model and a dif-
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Pharmacological Reports, 2012, 64,
643–649
ferent anesthetic agent. Another possibility lies in the
difficulty in detecting small differences with a fluidfilled catheter. The tip catheter has important properties that improve the sensitivity of hemodynamic
measurements. Conversely, the means from the three
groups are closely similar and seem to be true and not
due to the method used.
In summary, our results showed that acute administration of G-CSF can modulate cardiac remodeling
by preventing collagen deposition, myocyte hypertrophy and left ventricular dilation, contributing to
avoiding hemodynamic deterioration and heart failure
development. Further experiments need to be performed before these results can be used in clinical settings. However, it has been proven that G-CSF is safe
for human use, and thus, could help as a possible adjuvant therapy for heart failure induced by sympathetic overload.
Acknowledgments:
This study was supported by grants from National Council for
Scientific and Technological Development (CNPq) and Espírito
Santo Research Foundation (FAPES/PRONEX).
References:
1. Baldo MP, Davel AP, Damas-Souza DM, NicolettiCarvalho JE, Bordin S, Carvalho HF, Rodrigues SL
et al.: The antiapoptotic effect of granulocyte colonystimulating factor reduces infarct size and prevents heart
failure development in rats. Cell Physiol Biochem, 2011,
28, 33–40.
2. Baldo MP, Davel AP, Nicoletti-Carvalho JE, Bordin S,
Rossoni LV, Mill JG: Granulocyte colony-stimulating
factor reduces mortality by suppressing ventricular arrhythmias in acute phase of myocardial infarction in rats.
J Cardiovasc Pharmacol, 2008, 52, 375–380.
3. Baldo MP, Forechi L, Morra EAS, Zaniqueli D,
Machado RC, Lunz W, Rodrigues SL, Mill JG: Longterm use of low dose spironolactone in spontaneously
hypertensive rats. Effects on left ventricular hypertrophy
and stiffness. Pharmacol Rep, 2011, 63, 975–982.
4. Baldo MP, Rodrigues SL, Mill JG: Granulocyte colonystimulating factor for ischemic heart failure: should we
use it? Heart Fail Rev, 2010, 15, 613–623.
5. Brooks WW, Conrad CH: Isoproterenol-induced myocardial injury and diastolic dysfunction in mice: structural
and functional correlates. Comp Med, 2009, 59, 339–343.
6. Broskova Z, Knezl V: Protective effect of novel pyridoindole derivatives on schemia/reperfusion injury of the
isolated rat heart. Pharmacol Rep, 2011, 63, 967–974.
G-CSF prevents early cardiomyopathy in rats
Ludimila Forechi et al.
7. Carll AP, Willis MS, Lust RM, Costa DL, Farraj AK:
Merits of non-invasive rat models of left ventricular
heart failure. Cardiovasc Toxicol, 2011, 11, 91–112.
8. Dawn B, Guo Y, Rezazadeh A, Huang Y, Stein AB,
Hunt G, Tiwari S et al.: Postinfarct cytokine therapy regenerates cardiac tissue and improves left ventricular
function. Circ Res, 2006, 98, 1098–1105.
9. dos Santos L, Mello AF, Antonio EL, Tucci PJ: Determination of myocardial infarction size in rats by echocardiography and tetrazolium staining: correlation, agreements, and simplifications. Braz J Med Biol Res, 2008,
41, 199–201.
10. Feng W, Li W: The study of ISO induced heart failure rat
model. Exp Mol Pathol, 2010, 88, 299–304.
11. Grimm D, Elsner D, Schunkert H, Pfeifer M, Griese D,
Bruckschlegel G, Muders F et al.: Development of heart
failure following isoproterenol administration in the rat:
role of the rennin-angiotensin system. Cardiovasc Res,
1998, 37, 91–100.
12. Harada M, Qin Y, Takano H, Minamino T, Zou Y, Toko
H, Ohtsuka M et al.: G-CSF prevents cardiac remodeling
after myocardial infarction by activating the Jak-Stat pathway in cardiomyocytes. Nat Med, 2005, 11, 305–311.
13. Hou XW, Son J, Wang Y, Ru YX, Lian Q, Majiti W,
Amazouzi A et al.: Granulocyte colony-stimulating factor reduces cardiomyocyte apoptosis and improves cardiac function in adriamycin-induced cardiomyopathy in
rats. Cardiovasc Drugs Ther, 2006, 20, 85–91.
14. Kanellakis P, Slater NJ, Du XJ, Bobik A, Curtis DJ:
Granulocyte colony-stimulating factor and stem cell factor improve endogenous repair after myocardial infarction. Cardiovasc Res, 2006, 70, 117–125.
15. Li L, Takemura G, Li Y, Miyata S, Esaki M, Okada H,
Kanamori H et al.: Granulocyte colony-stimulating factor improves left ventricular function of doxorubicininduced cardiomyopathy. Lab Invest, 2007, 87, 440–455.
16. Li Y, Takemura G, Okada H, Miyata S, Esaki M,
Maruyama R, Kanamori H et al.: Treatment with granulocyte colony-stimulating factor ameliorates chronic
heart failure. Lab Invest, 2006, 86, 32–44.
17. Macambira SG, Vasconcelos JF, Costa CR, Klein W,
Lima RS, Guimarães P, Vidal DT et al.: Granulocyte
colony-stimulating factor treatment in chronic Chagas
disease: preservation and improvement of cardiac structure and function. FASEB J, 2009, 23, 3843–3850.
18. Mill JG, Stefanon I, dos Santos L, Baldo MP: Remodeling in the ischemic heart: the stepwise progression for
heart failure. Braz J Med Biol Res, 2011, 44, 890–898.
19. Minatoguchi S, Takemura G, Chen XH, Wang N, Uno Y,
Koda M, Arai M et al.: Acceleration of the healing process and myocardial regeneration may be important as
a mechanism of improvement of cardiac function and remodeling by postinfarction granulocyte colony-stimulating
factor treatment. Circulation, 2004, 109, 2572–2580.
20. Miyata S, Takemura G, Kawase Y, Li Y, Okada H,
Maruyama R, Ushikoshi H et al.: Autophagic cardiomyocyte death in cardiomyopathic hamsters and its prevention by granulocyte colony-stimulating factor. Am J
Pathol, 2006, 168, 386–397.
21. Nogueira BV, Palomino Z, Porto ML, Balarini CM,
Pereira TM, Baldo MP, Casarini DE et al.: Granulocyte
colony stimulating factor prevents kidney infarction and
attenuates renovascular hypertension. Cell Physiol Biochem, 2012, 29, 143–152.
22. Okada H, Takemura G, Li Y, Ohno T, Li L, Maruyama
R, Esaki M et al.: Effect of a long-term treatment with
a low-dose granulocyte colony-stimulating factor on
post-infarction process in the heart. J Cell Mol Med,
2008, 12, 1271–1283.
23. Orlic D, Kajstura J, Chimenti S, Limana F, Jakoniuk I,
Quaini F, Nadal-Ginard B et al.: Mobilized bone marrow
cells repair the infarcted heart, improving function and survival. Proc Natl Acad Sci USA, 2001, 98, 10344–10349.
24. Palojoki E, Saraste A, Eriksson A, Pulkki K, Kallajoki
M, Voipio-Pulkki LM, Tikkanen I: Cardiomyocyte apoptosis and ventricular remodeling after myocardial infarction in rats. Am J Physiol Heart Circ Physiol, 2001, 280,
H2726–2731.
25. Park C, Yoo JH, Jeon HW, Kang BT, Kim JH, Jung DI,
Lim CY et al.: Therapeutic trial of granulocyte-colony
stimulating factor for dilated cardiomyopathy in three
dogs. J Vet Med Sci, 2007, 69, 951–955.
26. Prabhu SD, Wang G, Luo J, Gu Y, Ping P, Chandrasekar
B: b-Adrenergic receptor blockade modulates Bcl-XS expression and reduces apoptosis in failing myocardium.
J Mol Cell Cardiol, 2003, 35, 483–493.
27. Ribeiro DA, Buttros JB, Oshima CT, Bergamaschi CT,
Campos RR: Ascorbic acid prevents acute myocardial
infarction induced by isoproterenol in rats: role of inducible nitric oxide synthase production. J Mol Hist,
2009, 40, 99–105.
28. Rona G: Catecholamine cardiotoxicity. J Mol Cell Cardiol, 1985, 17, 291–306.
29. Teerlink JR, Pfeffer JM, Pfeffer MA: Progressive ventricular remodeling in response to diffuse isoproterenolinduced myocardial necrosis in rats. Circ Res, 1994, 75,
105–113.
30. Wittstein IS, Thiemann DR, Lima JA, Baughman KL,
Schulman SP, Gerstenblith G, Wu KC et al.: Neurohumoral features of myocardial stunning due to sudden
emotional stress. N Engl J Med, 2005, 352, 539–548.
31. Yung MC, Hsu CC, Kang CY, Lin CL, Chang SL, Wang
JJ, Lin MT et al.: A potential for granulocyte colony
stimulating factor for use as a prophylactic agent for
heatstroke in rats. Eur J Pharmacol, 2011, 661, 109–117.
Received: July 14, 2011; in the revised form: January 6, 2012;
accepted: February 17, 2012.
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