54
Effect of Pravastatin on Phenotypical Transformation
of Fibroblasts and Hypertrophy of Cardiomyocytes
in Culture
O. M. Moiseeva, E. G. Semyonova*, E. V. Polevaya*,
and G. P. Pinayev*
Translated from Byulleten’ Eksperimental’noi Biologii i Meditsiny, Vol. 143, No. 1, pp. 60-63, January, 2007
Original article submitted July 28, 2006
We studied the effects of pravastatin on the functional state of cardiomyocytes and
cardiac fibroblasts in culture. Pravastatin in doses of 0.1-10.0 µM suppressed prolifera-
tion of cardiac fibroblasts and their phenotypical transformation into myofibroblasts and
prevented the development of cardiomyocyte hypertrophy. Pleiotropic effects of pra-
vastatin can be explained by inhibition of production transforming growth factor β
1
production.
Key Words: myofibroblasts; cardiomyocyte hypertrophy; statins
V. A. Almazov Institute of Cardiology, Russian Ministry of Health;
*Institute of Cytology, Russian Academy of Sciences, St. Petersburg.
Address for correspondence:
omoiseeva@front.ru. O. M. Moiseyeva
The problems of cardiomyocyte (CMC) hypertro-
phy and apoptosis are closely related to the de-
velopment of interstitial and perivascular fibrosis,
and hence, the type of structural pathological chan-
ges in the heart is more accurately denoted by the
term “myocardial remodeling”. Some drugs can
not only induce regression of myocardial hyper-
trophy, but also modify stromal elements of the
heart as a component of remodeling. However, the
cell aspects of these structural changes are not stu-
died.
We studied the effect of pravastatin (PS; hydro-
xymethylglutaryl-coenzyme A-reductase) on the
functional state of CMC and cardiac fibroblasts in
culture.
MATERIALS AND METHODS
Cardiomyocytes were obtained from the hearts of
2-4-day-old rat pups; cardiac fibroblasts were iso-
lated by differentiated adhesion. Fibroblasts of pas-
sages 3-6 were used in the study.
Control CMC were cultured in DMEM supple-
mented with 5% FCS; CMC hypertrophy was mo-
deled by adding 10% FCS to the medium.
Cardiomyocytes were visualized by EA53 mono-
clonal antibodies to α-actinin and rhodamine falloi-
din (Molecular Probes Inc.). VA type myofibro-
blasts were detected by immunofluorescent ana-
lysis using monoclonal antibodies to vimentin
(Sigma) and α-smooth-muscle actin (clone 1A4;
DAKO). Proliferative activity of cardiac fibroblasts
was evaluated by an indirect immunohistochemical
method with monoclonal antibodies to proliferating
cell nuclear antigen (PCNA) and visualized by the
LSAB2 system (DAKO). Immunofluorescent ana-
lysis of CMC and cardiac fibroblasts for transfor-
ming growth factor-β
1
(TGF-β
1
) was carried out
using original antibodies obtained at Department of
Cell Cultures, Institute of Cytology.
Apoptosis was detected by the method of neu-
tral single-cell DNA-comet assay and staining of
DNA fragments with DAPI fluorescent dye [2].
The degree of CMC hypertrophy in vitro was
evaluated by cytomorphometrical parameters and
Bulletin of Experimental Biology and Medicine, Vol. 143, No. 1, 2007 PHARMACOLOGY AND TOXICOLOGY
0007-4888/07/14310054 © 2007 Springer Science+Business Media, Inc.