Volume 02 Issue 05-2022
6
International Journal of Medical Sciences And Clinical Research
(ISSN
–
2771-2265)
VOLUME
02
I
SSUE
05
Pages:
06-13
SJIF
I
MPACT
FACTOR
(2021:
5.
694
)
(2022:
5.
893
)
OCLC
–
1121105677
METADATA
IF
–
5.654
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Oscar Publishing Services
Servi
ABSTRACT
The effect of plant proanthocyanidin – yantacin isolated from the camel thorn plant (Alhagi pseudalhagi) – Yantacin,
when administered orally at a dose of 100 mg / kg, on possible changes in the early stages of liver fibrosis that occurred
against the background of chronic heliotrine hepatitis in experimental conditions was studied. The studies were
conducted on male rats with a div weight of 100 ±10 g. The effect of yantacin on the general and biochemical
Research Article
STUDY THE EFFECT OF YANTACIN ON SOME INDICATORS OF CELLULAR
RENEWAL AND ON THE LEVEL OF PROTEIN EXPRESSION ON RAT
HEPATOCYTES IN CHRONIC HELIOTRINE LIVER DAMAGE
Submission Date:
May 09, 2022,
Accepted Date:
May 18, 2022,
Published Date:
May 30, 2022
Crossref doi:
https://doi.org/10.37547/ijmscr/Volume02Issue05-02
A.N. Aripov
Doctor of Medical Sciences, Professor, Republican Specialized Pediatric Research and Practice Medical Center,
Tashkent, Uzbekistan
O.A. Aripov
Doctor of Medical Sciences, Senior Research Fellow Republican Specialized Pediatric Research and Practice
Medical Center, Tashkent, Uzbekistan
L.L. Akhundjanova
Candidate of Biological Sciences, Senior Research Fellow Republican Specialized Pediatric Research and
Practice Medical Center, Tashkent, Uzbekistan
A.U. Nabiev
Junior Research Fellow Republican Specialized Pediatric Research and Practice Medical Center, Tashkent,
Uzbekistan
D.A. Nabieva
Doctor of Medical Sciences, Senior Research Fellow Republican Specialized Pediatric Research and Practice
Medical Center, Tashkent, Uzbekistan
T.T. Khamroev
Junior Research Fellow S.Yu. Yunusov Institute of Plant Chemistry, Tashkent, Uzbekistan
Journal
Website:
https://theusajournals.
com/index.php/ijmscr
Copyright:
Original
content from this work
may be used under the
terms of the creative
commons
attributes
4.0 licence.
Volume 02 Issue 05-2022
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International Journal of Medical Sciences And Clinical Research
(ISSN
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2771-2265)
VOLUME
02
I
SSUE
05
Pages:
06-13
SJIF
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MPACT
FACTOR
(2021:
5.
694
)
(2022:
5.
893
)
OCLC
–
1121105677
METADATA
IF
–
5.654
Publisher:
Oscar Publishing Services
Servi
parameters of blood, as well as on p53, PCNA of liver tissue, mitotic and apoptotic index against the background of
heliotrine liver damage was studied. As a result of the conducted studies, yantacin demonstrated a significant
protective effect compared with the control group on the detected fibrosis, according to which certain indicators,
against the background of chronic toxic liver damage. In this regard, a more extensive in-depth study of the protective
and restorative effects of the studied plant proanthosianide substance on chronic changes in the liver, serious
structural and functional disorders is being carried out.
KEYWORDS
Chronic hepatitis, liver fibrosis, liver cirrhosis, proanthocyanidin, p53, liver tissue PCNA, heliotrine lesion, yantacin,
hepatoprotective, antifibrotic.
INTRODUCTION
It is known that chronic liver diseases associated with
hepatitis B and C viruses are one of the urgent
problems of modern medicine. The incidence of
chronic hepatitis B and C in recent decades throughout
the world, including in Uzbekistan, has a steady
upward trend. Given the widespread prevalence of
chronic HBV and HCV infections among children, the
high frequency of adverse outcomes with disability and
significant mortality in the final stages of the natural
course, they represent one of the most pressing
problems of modern pediatrics. Chronic HBV infection
is observed in 5%-15% of patients with acute viral
hepatitis; liver cirrhosis develops in 10% of them, and
hepatocellular carcinoma forms in 8% of patients with
liver cirrhosis [1, 2, 3]. Recently, significant progress has
been made in the study of the pathogenesis of liver
fibrosis in CHB. The widespread spread of chronic liver
diseases, including hepatitis, fibrosis and insufficient
effectiveness for predicting the course of the disease
and timely prescribing their treatment require the
development of new approaches to the therapy of this
pathology. One of the promising directions in the
search for treatment of chronic hepatitis is associated
with the use of drugs, including those of plant origin,
with the ability to reduce the degree of fibrous changes
in the liver [4,5,6].
THE MAIN RESULTS AND FINDINGS
There are no effective methods of treating liver
fibrosis. This makes it urgent to search for new
methods aimed at the functional restoration of the
liver and its regeneration. In connection with this,
extensive pharmacological and toxicological and
chemical-technological studies of Yantacin plant
proanthocyanidin, which is isolated from the camel
thorn plant (Alhagi pseudalhagi), widely distributed in
Central Asia and Kazakhstan, were carried out in the
laboratories of the Research Institute of Pediatrics and
ICPS of the Academy of Sciences of the Republic of
Uzbekistan in a new direction [7, 8].
The above was the basis for determining the purpose
of this study and formulating tasks to achieve it.
Volume 02 Issue 05-2022
8
International Journal of Medical Sciences And Clinical Research
(ISSN
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2771-2265)
VOLUME
02
I
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05
Pages:
06-13
SJIF
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FACTOR
(2021:
5.
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(2022:
5.
893
)
OCLC
–
1121105677
METADATA
IF
–
5.654
Publisher:
Oscar Publishing Services
Servi
To evaluate the effectiveness of plant-derived yantacin
of hepatoprotective and antifibrotic action in chronic
toxic liver disease in rats.
The experimental part was performed on white
mongrel male rats. An experimental model of chronic
liver pathology was obtained in male rats weighing
100±10 g. The hepatotoxicity of heliotrin has been
shown by us in rats before, so the model of heliotrin-
induced hepatitis or liver fibrosis is the most suitable
model of toxic liver damage. The chronic form of
experimental
hepatitis
was
caused
by
the
administration of heliotrin in decreasing doses:
according to the scheme: 10 mg/100 g of mass, 7 mg/
100 g of mass, 5 mg/100 g of mass, 3 mg/ 100 g of mass
[9]. Chronic intoxication, confirmed morphologically,
was obtained on the 35th day of the experiment [10].
The mortality rate was 30%. On the 35th day of the
experiment, the animals were taken out of the
experiment under ether anesthesia, the liver was fixed
and poured into paraffin according to the standard
procedure. Also, to characterize the degree of fibrosis
of liver tissue, a general and biochemical blood
analysis, a morphological examination of the liver were
performed simultaneously.
To determine the level of apoptosis, special research
methods were used: determination of p53, PCNA of
liver tissue of experimental animals was carried out by
immunohistochemical method (ShiSh.R., 1991) [8].
With the introduction of heliotrin in the blood of
experimental animals, a change in several indicators
was detected; leads to a positive dynamics of
morphological data, causes the reverse development
of hyperfermentemia and hyperbilirubinemia; a
significant decrease in cytokine levels; levels
hyperlipidemia and hypoproteinemia; there is a
tendency to normalize not only qualitative, but also
quantitative
indicators
of
erythropoiesis:
the
hematocrit index decreases, necrosis and fibrosis
decreases.
It is known that apoptosis is triggered either through
receptors located on the surface of hepatocytes, or
through structural and metabolic disorders in the
mitochondria of cells. The receptor pathway of
triggering apoptosis occurs due to the interaction of
pro–apoptotic molecules (FasL ligand) with "death
receptors" (FasR). Namely, the formation of a ligand
receptor complex (FasR/FasL) contributes to the
further launch of a cascade of intracellular reactions
that ultimately ensure the realization of apoptotic
death of hepatocytes. In the conditions of our
experiments, the hepatocyte is influenced by a toxic
factor – heliothrin, which can contribute to damage to
the plasma membrane, including its receptor
apparatus and, subsequently, disruption of ligand–
receptor relationships. Although the main factors
released by mitochondria into the cytoplasm are
cytochrome C and apoptosis –indicating factor (AIF), as
well as procaspases 2, 3 and 9, which trigger
intracellular apoptosis and participate in many
signaling pathways that ensure cell death, but this
does not always end with the realization of cell
apoptosis. Apoptosis and necrosis in the pathogenesis
of experimental chronic liver damage. To assess the
state of cellular renewal in conditions of chronic liver
damage by heliotrin, we determined in the liver tissue
of experimental animals the indicators of apoptotic
index (AI) and hepatocyte necrosis as two main ways
of cell death, as well as the mitotic index (MI) – an
indicator of liver proliferative activity. In addition, the
coefficients of the ratio of apoptosis to proliferation
(AI/MI) and the ratio of apoptosis to necrosis were
analyzed (Table 1).
Volume 02 Issue 05-2022
9
International Journal of Medical Sciences And Clinical Research
(ISSN
–
2771-2265)
VOLUME
02
I
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05
Pages:
06-13
SJIF
I
MPACT
FACTOR
(2021:
5.
694
)
(2022:
5.
893
)
OCLC
–
1121105677
METADATA
IF
–
5.654
Publisher:
Oscar Publishing Services
Servi
Table 1. Some indicators of cellular renewal in the liver in chronic heliothrin hepatitis against the background
of the effect of Yantacin. (M±m).
Animal groups
Indicators
МI (mitotic
index, per %)
АI (apoptotic
index, per %)
Necrosis
(per %)
Coefficients
АI/МI
Necrosis /АI
Control
4,2±0,30
6,10±0,25
--
1,2±0,12
--
Chronic heliotrical
hepatitis
1,9±0,46*
10,2±0,32*
52,2±0,92
5,4±0,25*
5,1±0,36
Acute heliotrical
hepatitis
2,4±0,40*
6,4±0,21*
58,2±1,51
2,7±0,21*
9,1±1,24
Chronic heliotrical
hepatitis + Yantacin
3,1±0,32*
8,70,21*
33,1±1,24
2,81±0,45
3,81±0,92
Notes: - differences with intact ones are significant * (p<0.05).
Consequently, in conditions of chronic heliotrine
hepatitis, the apoptotic death of hepatocytes
increases, but, clearly, the death of hepatocytes due to
necrosis increases almost 7 times. Such a pathway of
liver cell death is practically not found in animals of the
control group. Despite the high frequency of cell death
(approximately more than 60 hepatocytes out of 100 in
the field of vision are susceptible to death), the mitotic
activity (MI) of hepatocytes is low -2.2 times lower than
that in the control group (Table. 1), and the coefficient
AI/MI, on the contrary, is 4.5 times higher.
Consequently, under conditions of prolonged
exposure to relatively low doses of hepatotropic toxic
factor, i.e., with chronic heliothrin hepatitis, there is a
high level of hepatocyte death relative to the norm, but
in intensity it gives way to necrosis, and the processes
aimed at restoring hepatocyte deficiency not only do
not have time to compensate for the missing cells, but
even noticeably decrease, about as evidenced by the
low mitotic activity of the liver and the high value of the
AI coefficient/MI, there is a noticeable activation of
hepatocyte apoptosis and necrosis. At the same time,
AI in chronic heliotrine hepatitis is 1.7 times higher than
the control value.
Under the conditions of HCG, necrosis exceeds
apoptosis by 5 times, i.e. if 5 out of 6 cells subject to
death undergo necrosis, then 1 undergoes apoptosis.
Consequently, in conditions of chronic heliotrine
hepatitis, the apoptotic death of hepatocytes
increases, but, clearly, the death of hepatocytes due to
necrosis increases almost 7 times. Such a pathway of
liver cell death is practically not found in animals of the
control group. Despite the high frequency of cell death
(more than 60 hepatocytes out of 100 in the field of
vision are susceptible to death), the mitotic activity
Volume 02 Issue 05-2022
10
International Journal of Medical Sciences And Clinical Research
(ISSN
–
2771-2265)
VOLUME
02
I
SSUE
05
Pages:
06-13
SJIF
I
MPACT
FACTOR
(2021:
5.
694
)
(2022:
5.
893
)
OCLC
–
1121105677
METADATA
IF
–
5.654
Publisher:
Oscar Publishing Services
Servi
(MI) of hepatocytes is low. The positive effect of the
studied substance on the apoptotic index, mitotic
activity (MI) of hepatocytes, in this regard, the AI
coefficient/MI and Necrosis/AI against the background
of chronic heliotrine hepatitis in rats is presented in
Table 1.
Based on the above, we studied the expression of
protein p 53 on hepatocytes in conditions of chronic
heliotrine hepatitis. The results of these studies are
presented in table 2. The results of the conducted
studies show that under conditions of toxic liver
damage, certain shifts in gene regulation occur, in
particular through the p53 protein that characterizes
hepatocyte apoptosis.
Table 2. Expression of p53 protein on hepatocytes in conditions of acute and chronic heliotrine hepatitis and
against the background of the effect of Yantacin. (М±m)
The studied groups of
animals
Number of
samples
Positive expression of p53 protein
Number of positive
observations
Frequency of p 53 –
positive cells, per %
Control
10
2
8,2±0,52
Chronic heliotrical hepatitis
10
5
14,11±1,09*
Chronic heliotrical hepatitis +
Yantacin
10
4
11,24±1,21*
Notes: - differences with intact ones are significant * (p<0.05)
At the same time, it was found that if in healthy animals
hepatocytes with a positive reaction to p 53 are found
in 20% of animals, then with ChHH – in 50% of animals.
Along with this, the level of p53 protein expression on
the hepatocytes of experimental animals is also higher
than that of the control (Table 2). The expression of
p53 in ChHH exceeds the control value by 71.9%.
Consequently, in conditions of toxic liver damage, not
only an increase in the number of animals with a
positive reaction to p53 occurs, but the expression of
this protein on hepatocytes is noticeably increased,
and this is clearly seen in animals under prolonged
exposure to a toxic agent, i.e. with chronic heliotrine
hepatitis.
If this process proceeded in a balanced manner with
the process of cellular renewal, i.e. with proliferation,
then indeed, in such a change in the death of
hepatocytes, as in ChHH, one could see elements of
rationality. However, in the conditions of ChHH, the
opposite picture is observed, characterized by further
inhibition of mitotic activity, which increases to an even
greater extent the imbalance between the death of
hepatocytes and their renewal. Apparently, this is the
essence of the complexity of the pathogenesis of
chronic liver pathology, its progression, as well as the
formation of the process of fibrogenesis with the
transition to cirrhosis.
Volume 02 Issue 05-2022
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OCLC
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IF
–
5.654
Publisher:
Oscar Publishing Services
Servi
Proliferation, as a compensatory process, allows under
normal conditions to maintain homeostasis with
natural cell renewal. In conditions of pathology,
proliferation, which has complex mechanisms of
regulation, is aimed at regenerating damaged tissue
and maintaining its functional viability. We evaluated
the proliferative activity of the liver parenchyma
(hepatocytes) and one of the elements of its stroma
(sinusoid cells) in conditions of ChHH. The positive
effect of the studied substance on the level of
expression of the p53 protein on hepatocytes against
the background of chronic heliotrine hepatitis in rats is
presented in Table 2.
It is known that apoptosis is triggered either through
receptors located on the surface of hepatocytes, or
through structural and metabolic disorders in the
mitochondria of cells. The receptor pathway of
triggering apoptosis occurs due to the interaction of
pro–apoptotic molecules (FasL ligand) with "death
receptors" (FasR). Namely, the formation of a ligand-
receptor complex (FasR/FasL) contributes to the
further launch of a cascade of intracellular reactions
that ultimately ensure the realization of apoptotic
death of hepatocytes. In the conditions of our
experiments, the hepatocyte is influenced by a toxic
factor – heliothrin, which can contribute to damage to
the plasma membrane, including its receptor
apparatus and, subsequently, disruption of ligand–
receptor relationships.
How can we explain the low level of proliferative
activity of hepatocytes detected by us in the conditions
of chronic liver damage by heliotrin Indeed, in
conditions of increased hepatocyte death, an increase
in the rate of proliferation should also occur in order to
maintain the balance of functioning hepatocytes.
However, this does not happen. To clarify this issue, we
studied the expression of another protein, PCNA, on
hepatocytes in conditions of toxic liver damage, since
this protein is widely used to assess the proliferative
activity of cells. The results of the conducted research
in this direction are presented in Table 3.
Table 3. Expression of PCNA protein on hepatocytes in conditions of acute and chronic heliotrine hepatitis
and against the background of the effect of Yantacin. (М±m)
The studied groups of
animals
Number of
samples
Positive expression of PCNA protein
Number of positive
observations
Frequency of PCNA –
positive cells, per %
Control
10
10
54,2±3,1
Chronic heliotrical hepatitis
10
6
9,7±1,28*
Chronic heliotrical hepatitis +
Yantacin
10
8
29,1±1,3
Notes: - differences with intact ones are significant * (p<0.05)
Volume 02 Issue 05-2022
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06-13
SJIF
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(2021:
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(2022:
5.
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OCLC
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METADATA
IF
–
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Publisher:
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Servi
As can be seen from the presented data, in conditions
of chronic heliotrine hepatitis, noticeable changes in
PCNA expression occur.
At the same time, 60% of animals with HCG have a
positive reaction in hepatocytes to the PCNA protein,
At the same time, in animals of the control group, this
was the case in all animals (100%). Analysis of the
results of PCNA protein expression on hepatocytes of
experimental animals showed that it was also
noticeably lower than in animals of the control group
(Table 3). At the same time, if the expression of this
protein on hepatocytes of animals with ChHH, it was
5.6 times lower than in the control group.
Consequently, under conditions of toxic liver damage,
there is a significant decrease in the expression of the
hepatocyte proliferation marker on hepatocytes.
Indeed, the morphological picture of the liver, studied
by us in chronic heliotrine hepatitis, also indicates that
in the conditions of the studied liver pathology, the
structural changes characteristic of chronic hepatitis
differ only in isolated cases - these are hepatocytes
with signs of mitosis and, consequently, proliferation.
The positive effect of the studied substance on the
level of PCNA protein expression on the hepatocytes
of experimental animals against the background of
chronic heliotrine hepatitis in rats is presented in Table
3.
CONCLUSION
Thus, the obtained results showed that plant
proanthocyanidin significantly reduced the death of
hepatocytes and accelerated their regeneration in
small quantities. In this regard, in order to recommend
the use of yantacin for chronic liver diseases,
administered for therapeutic purposes with toxic
hepatitis, a more extensive in-depth study of the
protective and restoring effect of the studied
substance of plant proanthosianide on chronic
changes in the liver, serious structural and functional
disorders is carried out.
REFERENCES
1.
Кузнецова А.В., Жердева А.И., Горовенко
Н.А. и др. Место и роль серологических
маркеров фиброза в диагностике и лечении
хронической
HCV-инфекции
//
Клин.
Перспективы гастроэнтерол, гепатол. —
2007. — № 3. — С. 35-39.
2.
Лобзин, Ю.В. Вирусные поражения печени:
от болезни Боткина до современной
инфекционной гепатологии / Ю.В. Лобзин,
К.В.Жданов // Медицинский академический
журнал – 2012. - №2, Том 12, - С.7-18
3.
Мицура, В.М. HCV-ассоциированный цирроз
печени: клинические проявления и причины
неблагоприятных исходов / В.М. Мицура //
Клиническая инфектология и паразитология.
– 2013. – №3. – С. 75-82.
4.
Абдурахманов,
Д.Т.Противовирусная
терапия и регресс фиброза печени при
хроническом гепатите В / Д.Т.
Абдурахманов
//
Росc.
журн.
гастроэнтерол., гепатол., колопроктол. –
2010. - №1. – С.14-20.
5.
Aripov A.N., Aripov O.A., Akhundjanova L.L.,
Nabiev A.U., Nishonboev S.Z., Nabieva D.A.,
Akhmedova D.I., & Khamroev T.T.. (2022).
Study of the hepatotropic activity of
saforoflavonoloside
under
experimental
conditions. Oriental Journal of Medicine and
Pharmacology,
2(02),
55–64.
https://doi.org/10.37547/supsci-ojmp-02-02-07
6.
Шипунов М.В. Иммунные предикторы
фиброгенеза и их использование для
неинвазивной диагностики фиброза печени
у больных хроническими вирусными
Volume 02 Issue 05-2022
13
International Journal of Medical Sciences And Clinical Research
(ISSN
–
2771-2265)
VOLUME
02
I
SSUE
05
Pages:
06-13
SJIF
I
MPACT
FACTOR
(2021:
5.
694
)
(2022:
5.
893
)
OCLC
–
1121105677
METADATA
IF
–
5.654
Publisher:
Oscar Publishing Services
Servi
гепатитами / Шипунов М.В. [и др.] //
Бюллетень ВСНЦ СО РАМН. – 2012. – Т.85,
№3. – С.231-236.
7.
Zou G.A., Mansur S., Hu S.C., Aisa H.A.,
Shakhidoyatov Kh.M. Chemistry of Natural
Compounds, 2012, vol. 48 (4),pp. 635–637.
8.
Фесенко Л.М., Арипов А.Н..Ахунджанова Л.Л.
Экспериментальное
обоснование
антифиброзной эффективности янтацина в
печени крыс с хронической гелиотриновой
интоксикацией.
Научно-практический
журнал Педиатрия №3/2019. С.211-215.
9.
Абдуллаев Н.Х., Каримов Х.Я. Печень при
интоксикациях гепатотропными ядами. –
Ташкент: Медицина, 1989. – 98 с.
10.
Aripov A.N, Akhunjanova L.L, Khamroev T.T,
Aripov Abdumalik Nigmatovich, Akhunjanova
Lola
Lazizovna,
&
Khamroev
Tolmas
Tolibovich. (2022). Differential Analysis of
Chronic Toxic Hepatitis Caused by The
Introduction of Heliotrin Solution in Various
Ways. Texas Journal of Medical Science, 4, 58–
62.
Retrieved
from
https://zienjournals.com/index.php/tjms/articl
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