53 / 103
I
mplications
of
alcoholic
cirrhosis
in
atherosclerosis
of
autopsied
patients
R
ev
A
ssoc
M
ed
B
ras
2017; 63(4):336-340
337
endothelial function, limiting the effective diameter of
the vessels, and can also cause thrombosis, which can
trigger various kinds of strokes, such as sudden death
from myocardial infarction.
8
Several risk factors have been described for athero-
genic development, such as dyslipidemia, hypertension,
diabetes, smoking, alcohol abuse and physical inactivity.
Lipid profile has been widely studied for its strong asso-
ciation with atherogenesis.
9
The most characteristic com-
ponent of atherosclerosis is chronic inflammation of the
artery walls, pointing out the importance of inflammation
for the severity of the disease.
10
Regarding the advances in diagnostic and therapeutic
areas, cardiovascular disease remains a major cause of mor-
tality worldwide
11
and, in view of the epidemiological pro-
file of the Brazilian population, further research is essential
to identify risk groups and enable interventions to prevent
the incidence and progression of these diseases. In addition,
current treatment methods must be improved, seeking new
therapeutic targets and specific biomarkers that prove to
be useful in the medical practice. Classic research related
to the circulatory systemmainly involves histopathologic
descriptions of pathophysiological processes.
11
Our study aimed to quantify collagen fibers in the
aorta and liver of autopsied patients with and without
alcoholic cirrhosis, and to associate the percentage of
these fibers with the degree of atherosclerosis, age and
BMI of patients, in order to validate the hypothesis that
patients with cirrhosis have a higher degree of atheroscle-
rosis and higher percentage of collagen fibers both in the
liver and in the aorta.
M
ethod
This study was approved by the Ethics Committee of the
Federal University of Triângulo Mineiro (UFTM, in the
Portuguese acronym), under number 888.
The sampling comprised 30 samples of aorta and 30
of liver from autopsies routinely performed at the Clinical
Hospital of the Federal University of Triângulo Mineiro,
in the city of Uberaba, State of Minas Gerais, between 1983
and 2008. The information and the fragments were col-
lected by two pathologists responsible for the autopsies
performed by the Division of General Pathology of the
Clinical Hospital of the UFTM.
Based on autopsy reports, 15 patients were selected,
aged over 18 years and with alcoholic liver cirrhosis. For
the control cases, 15 patients without alcoholic cirrhosis
were selected, matched for gender, color and age and with
no cardiovascular disease, autopsied at the same period.
Thirty aortic fragments were removed transversely, mea-
suring about 20 x 1 mm, and 30 liver fragments of the
right lobe were collected, with approximately 2 x 2 cm.
As a contributor to the nutritional assessment of pa-
tients, the body mass index (BMI) was calculated using
the following formula: weight (kg)/height
2
(m). Patients
were classified according to BMI as underweight (BMI less
than 18.5 kg/m
2
), normal (between 18.5 and 24.99 kg/m
2
),
overweight (between 25.0 and 29.99 kg/m
2
) or obese (over
30.0 kg/m
2
). For this classification, the standards of the
World Health Organization were applied (WHO, 2004).
Macroscopic analysis of the aorta
Macroscopic degree of atherosclerosis was classified into
mild, moderate or severe. From this criterion, the obser-
vers of this study described quantitatively the atheroscle-
rotic arteriosclerosis. For this evaluation, the extent of
atheromatous plaques and the intensity of fibrosis and
calcification were considered.
12
The aortas were evaluated with the aid of a standard
scale of 12.0 cm. Each examiner subjectively measured
the degree of atherosclerosis using a non-millimetric scale,
marking a point on the scale. Next, with a ruler, the dis-
tance from 0.0 to the point marked on the scale was mea-
sured. The observers standardized atherosclerosis, accord-
ing to its intensity, categorizing as mild from 0.1 cm to
4.0 cm, moderate from 4.1 cm to 7.0 cm, and severe from
7.1 cm to 12.0 cm.
12
Microscopic analysis of aorta and liver fragments
The slides were stained with hematoxylin and eosin (HE)
for semi-quantitative evaluation of lipid deposits, analy-
zed under common light microscopy. After this analysis,
the degree of lipid deposit was classified as mild, when
the involvement of the section analyzed was less than 25%;
moderate from 26 to 50%, and severe if greater than 50%.
The morphological evaluation method of the aortas was
adapted from other studies.
13,14
Slides stained with picro-sirius were used for quantifi-
cation of collagen fibers in the inner layer of the aorta (Fig-
ures 1 and 2) and throughout the extension of the liver cut,
examined under polarized light with a 20x objective, final
magnification of 800x. The analyses of collagen fibers were
performed by morphometric software using the automat-
ic image analysis system KS-300
®
(Kontron-Zeiss).
Statistical analyses
Initially, a Microsoft Excel
®
spreadsheet was produced
and data were analyzed using Graphpad Prism
®
5.0 soft-
ware. The normality of the data was checked with Kol-
mogorov-Smirnov test. Since the distribution was non-