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Kamalov
et al
. European Journal of Pharmaceutical and Medical Research
17
STATE OF THE CYTOKINE PROFILE AND APOPTOSIS FACTORS OF CHILDREN
WITH DILATED CARDIOMYOPATHY
Kamalov Z. S.*, Aripova Sh. Sh., Achmedova D. I. and Sabirova F. B.
Institute of Human Immunology and Genomics, Academy of Sciences of the Republic of Uzbekistan Tashkent
Pediatric Medical Institute.
Article Received on 24/03/2020 Article Revised on 13/04/2020 Article Accepted on 03/05/2020
INTRODUCTION
In recent years, a wide study of apoptosis processes in
the development of pathological processes has begun.
This problem is relevant, reveals the importance of the
activation
of
apoptosis
factors
in
dilated
cardiomyopathies (DCMP) in children. DCMP is a
disease characterized by dilatation of the left ventricle
with the development of systolic heart failure and is not
associated with coronary heart disease or valvular
dysfunction.
[3,4,9,12]
Apoptotic cell death is known to be a fundamental
process in regulating immune responses. The classic
specific apoptosis induction receptor is CD95 + (Fas /
Apo-1). When cells are activated, CD95 + is most
expressed on neutrophils, CD4 + T-lymphocytes, which
characterizes their high sensitivity to FasL-induced
apoptosis, but the action of inducers is necessary to start
the process.
[1,5]
The pro-inflammatory cytokines TNF-α,
which is the main pro-apoptogenic cytokine and, at the
system level, leads to the induction of apoptosis of blood
cells, act as factors inducing apoptosis. Another inducing
factor is interleukin-18 (IL-18), which, affects the
secretion of IFNγ, activates the cells of the monocyte-
macrophage system, which leads to the activation of
many antibacterial and antiviral responses. It is believed
that the stress-induced release of IL-18 can lead to an
increase in the IFNγ cycle, which is involved in the
activation of the expression of Fas proteins (CD95 +),
the stimulation of which leads to apoptosis.
[2,6,7]
The aim of this
study was to study the state of the
cytokine profile and apoptosis factors in schoolchildren
with DCMP.
MATERIAL AND METHODS
The present study included 50 schoolchildren from 7 to
14 years old with an established diagnosis of DCMP.
The control group consisted of 20 children of the same
age.
Immunological studies in the examined children were
carried out in the laboratory of immunoregulation of the
Institute of Human Immunology and Genomics,
Academy of Sciences of the Republic of Uzbekistan.
The expression of CD95 + receptors was performed
using monoclonal antibodies produced by Sorbent LLC,
Russia (Moscow).
The concentration of tumor necrosis factor-α and
interleukin 18 (TNF-α and IL-18) in the blood serum was
determined
by
the
method
of
enzyme-linked
immunosorbent
assay
using
test
systems
ZAO
"VECTOR-BEST" (Russia, Novosibirsk). Quantitative
assessment of the results was carried out by constructing
a calibration curve or using the commercial computer
program
―Microplate
manager‖,
reflecting
the
dependence of optical density on concentration for a
standard antigen and allowing comparison of the studied
samples with it.
Statistical processing of the obtained data was carried out
using the computer program Statistica 6.0. The
SJIF Impact Factor 6.222
Research Article
ISSN 2394-3211
EJPMR
EUROPEAN JOURNAL OF PHARMACEUTICAL
AND MEDICAL RESEARCH
ejpmr, 2020,7(6), 17-19
ABSTRACT
A clinical and immunological examination of 21 school children from 7 to 14 years old with dilated
cardiomyopathy (DCMP) was carried out. As a control, 20 practically healthy children of comparable age were
examined. Significant changes in the studied parameters of the immune system were found in children with DCMP.
A significant increase in the concentration of pro-inflammatory (TNF-α and IL-18 increased by 4.2 and 3.0 times)
cytokines was revealed. The number of expressing apoptosis antigen - CD95 + cells in peripheral blood rises 2.0
times.
KEYWORDS:
children, dilated cardiomyopathy, immunity, lymphocytes, apoptosis, cytokines.
*Corresponding Author:
Kamalov Z. S.
Institute of Human Immunology and Genomics, Academy of Sciences of the Republic of Uzbekistan Tashkent Pediatric Medical Institute.
www.ejpmr.com
Kamalov
et al
. European Journal of Pharmaceutical and Medical Research
18
significance of differences in the average values of the
compared indicators was evaluated by Student's criterion
(t)
RESULTS AND DISCUSSION
Apoptosis, often called physiological cell death, is an
energetically active, genetically controlled process that
serves to eliminate defective or damaged cells. Apoptosis
helps preserve the order and normal functioning of the
biological system, cleansing of unclaimed, sick (who
have completed their life cycle or resulting from
mutations of potentially dangerous) cells and is a
fundamental process of maintaining homeostasis: both an
increase and a decrease in the level of apoptosis lead to
disruption of homeostasis and the development of
various diseases. IL-18 and apoptosis expressing antigen
- CD95 + lymphocyte cells are involved in the process of
initiating programmed cell death.
Interleukin 18, being a pleiotropic pro-inflammatory
cytokine, stimulates the production of IFNγ, TNFα, IL-1,
IL-2, apoptosis factors, increases the proliferative
activity of T-lymphocytes, increases the lytic activity of
NK cells. IL-18 is involved in the formation of cellular
and humoral
[6,7,8]
, innate and acquired immune
responses
[14]
, stimulates the production of adhesion
molecules that are involved in the mechanisms of cell
migration, which is important both in the formation of
the immune response and in pathogenesis of certain
diseases.
[13]
IL-18 not only stimulates the synthesis of INFγ, but also
modulates its functional activity. It was shown that the
expression of the Fas ligand of CD4 + Th1 and NK cells
also occurs under the influence of IL-18. On the other
hand, it was shown that INFγ is involved in the
activation of expression of Fas itself. Thus, we can
conclude that IL-18 alone (FasL) or through INFγ (Fas)
stimulates the initialization of apoptosis.
[8]
In the peripheral blood serum of healthy children, the
level of IL-18 ranged from 27 to 68 pg / ml, and
averaged 48.3 ± 2.84 pg / ml. A study of the production
of IL-18 showed a 3.0-fold increase in children with
DCMP (146.3 ± 9.00 pg / ml, P <0.001) in comparison
with the control group (Table 1.).
Table 1: The number of CD95 cells and the
concentration of certain cytokinesin children, (M
m).
Indicators
Control
Group n=20
DCMP
n=21
P
IL-18 pg/ml
48,3±2,84
146,3±9,00 P<0,001
TNF-α pg/ml
12,5±0,96
52,3±2,70
P<0,001
CD95 %
18,8±0,91
38,5
1,44
P<0,001
An increase in the concentration of IL-18 can lead to
activation of the expression of Fas proteins (CD95), the
stimulation of which leads to apoptosis.
The binding of soluble and surface-expressed activated
lymphocyte receptors (FasL- and Fas-) causes cell
apoptosis. The number of peripheral blood-expressing
apoptosis antigen - CD95 + cells in healthy children
averaged 18.8 ± 0.91% with individual values from 11 to
24%. This indicator in children with DCMP was
significantly increased (38.5 ± 1.44% P <0.001)
compared with those of healthy newborns.
An increase in the expression of apoptosis antigen -
CD95 + cells in the peripheral blood, which refers to a
membrane or receptor-mediated factor, initiates the
development of apoptosis. Through the C-terminal
intracellular domain of this receptor (the so-called death
domain), the implementation of the apoptogenic signal is
activated. The determination of CD95 + on the surface of
lymphocytes is regarded as their readiness for apoptosis.
The next stage of our research was to determine the level
of TNF-α in blood serum in children.
We conducted a study to determine the level of
production of TNF-α as an important mediator, which is
one of the most universal regulators of immunity and
inflammatory reactions with a wide range of biological
effects. The main producers are monocytes and
macrophages. It is also secreted by neutrophils,
endothelial and epithelial cells, eosinophils, mast cells, B
and T lymphocytes when they are involved in the
inflammatory process.
[7,8]
TNF-α normally plays a fundamental physiological role
in immunoregulation, but in some cases it can exert a
pathological effect, taking part in the development and
progression
of
inflammation,
microvascular
hypercoagulation,
hemodynamic
disturbances
and
metabolic depletion (cachexia) in various human
diseases, both infectious and non-infectious nature
[10]
,
including heart disease. A direct relationship was
established between TNF-α and heart failure syndrome,
which means that the level of TNF-α in the serum of
patients with severe heart failure is an order of
magnitude higher than in healthy individuals.
[11,15]
An analysis of the results showed that in healthy
individuals, individual indicators of TNF-α production
ranged from 7 to 20 pg / ml, and the average value of this
cytokine was 12.5 ± 0.96 pg / ml. Thus, the level of the
pro-inflammatory cytokine TNF-α was significantly
increased in the main group of patients (P <0.001) and
averaged 52.3 ± 2.70 pg / ml.
The ratio of the increase in the concentration of TNF-α
cytokine relative to the control values in patients with
DCMP was 4.2 times. The data obtained suggest that
there is a certain dependence of the level of TNF-α
concentration on the nature of the pathological process,
as evidenced by the very high level of its production in
the main group.
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Kamalov
et al
. European Journal of Pharmaceutical and Medical Research
19
Our studies have shown that in children with DCMP
there is a significant increase in the concentration of pro-
inflammatory (TNF-α and IL-18 increase by 4.2 and 3.0
times) cytokines. The number of expressing apoptosis
antigen - CD95 + cells in peripheral blood rises 2.0
times.
The level of TNF-α and IL-18 can indirectly judge the
activity of the inflammatory process as a whole. It is
likely that there is a positive correlation between the
severity of cardiac pathology and the level of synthesis
of TNF-α and IL-18, and the heavier the heart failure, the
stronger the response of the immune system and the
higher the level of cytokines, and vice versa.
Our data suggest that as a result of a critical increase in
the level of circulating pro-inflammatory cytokines, one
after another processes that have negative cardiovascular
effects are activated, which subsequently contribute to
even greater damage to the myocardium.
Thus, elevated levels of TNF-α, as well as IL-18
progressively have a direct damaging effect on
cardiomyocytes, inducing their apoptosis. Given the
great importance of mediators in providing the entire
mechanism for protecting the div from infectious and
other antigens, it can be considered highly relevant to
conduct a study of the level of pro-inflammatory
cytokines in heart failure in children with DCMP.
CONCLUSIONS
1. It was revealed that in children with DCMP, the level
of
pro-inflammatory
cytokines
was
significantly
increased, and this fact allows us to judge the activity of
the inflammatory process, which progressively has a
direct damaging effect on cardiomyocytes. So, in
children of the main group, the synthesis of TNF-α and
IL-18 is increased by 4.2 and 3 times, respectively, than
in children of the control group.
2. It has been established that in children with DCMP, a
significant (2.0-fold) increase in the number of CD95 +
cells expressing apoptosis antigen occurs in peripheral
blood.
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