Pre-Clinical Study of General Toxicity of the
Medication “Fitin-S”
Kamola Mirzaakhmedova
1
, Shakhida Ziyayeva
1
,
Abdushukiir Yuniisov
1
, Gidchekhra Karimova
1
, Rustam Karimov
1
1
Associate Professora. Tashkent Pediatric Medical Institute, Uzbekistan, Tashkent
Abstract
A pre-clinical study of the general toxicity of the medication “Fitin-S” has shown that it is referred to class IV of low toxic
compounds. The medication “Fitin-S” does not have aaccumulative and local irritant action. At multiple intramuscular
administration to rats and rabbits it does not influence the behavior and the dynamics of animals growth, does not cause a
toxic effect on the composition of peripheral blood, kidneys’ and liver’s function as well as on the pathomorphology of
animals organs and tissues. There is a slowdown in the process of blood coagulation in rabbits at doses of 16 and 32 mg/kg
during the whole study period. In 1 month of a recovery period all the indicators have been changing within physiological
norm. All the aforementioned data allows us to make a conclusion that the medication does not cause a toxic effect on the
organism of animals.
Keywords:
General toxicity, medication “Fitin-S”, acute toxicity, cumulation, chronic toxicity, local irritant action,
pathomorphological study.
Introduction
The purpose of studying the medication “Fitin-S” is to
establish the nature and severity of its damaging effects on
the div of experimental animals and assess its safety.
Materials and Method
The study of acute toxicity of the medicationshas been
carried out according to the method of Litchfield and
Wilcoxon on white mice, both sexes, weighing 20±2.0 g of
6 animals in each group, 150 mice has been used in total
2
.
All pharmacological studies have been performed on healthy
sexually mature animals (mice) exposed to the quarantine of
at least 10-14 days
1
.
The medication tested has been injected intravenously,
subcutaneously and intramuscularly as a single dose of 100
to 10000 mg/kg. The animals were monitored hourly during
the first day of the experiment in the laboratory, while the
survival rate during the experiment, general condition,
possible convulsions and death were used as indicators of
the functional state of the animals. At the end of the
experiment, the average lethal dose (LD50) has been
calculated and the toxicity class has been determined
3
. The
study of accumulation of “Fitin-S” has been carried out
according to the Lim’s method, which allows to evaluate not
only accumulation, but also addiction. The objective of the
study was to identify a possible accumulative property in the
solution of the medication tested. The trials have been
performed on 10 mice of both sexes weighing 20±2.0 g. The
medications have been administered intramuscularly
according to the scheme for 24 days from 0.1 to 1.15 from
LD
50
4
.
The general toxic action of “Fitin-S” at multiple
intramuscular administration has been studied in 40 non-
linear white rats (males and females) with the initial weight
of 110±10 grams and in 16 gray-colored rabbits of both
sexes with the initial weight 2,0±0,2 kilograms. Animals
were in vivarium on a normal diet. The trial animals have
been divided into 4 groups: 10 rats (5 males and 5 females)
and 4 rabbits (2 females and 2 males) in each. The
medication has been administered intramuscularly daily (7
days a week) for 30 days to rats at doses of 20, 40 and 100
mg/kg and to rabbits at 8.0, 16.0 and 40 mg/kg. The control
animals have been
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Medico-legal Update, October-December
2020, Vol. 20, No. 4
injected with distilled water in a
bioequivalent volume. All animals have been kept in
standard vivarium conditions on a full-fledged food and
water diet.
The condition of animals during chronic intramuscular
administration has been evaluated by the following
parameters of peripheral blood: haemoglobin contents, gr/dl;
the number of red blood cells,10
12
/l; reticulocytes,%
0
;
thrombocytes, 10
9
/l; leucocytes, 10
9
/ land leukogram; liver
function evaluation - by glucose content,mmol/1; total
protein,gr/dl; alanine - and aspartate - aminotransferases
(A1AT, AsAT) in blood serum. Biochemical tests have been
performed using test kits from CypressDiagnostica,
Belgium. Renal function has beenevaluated by dimesis with
a water load (5% of div weight) for 4 hours, blood urea,
urine acidity (pH), the presence of nitrites, glucose,
leukocytes etc. in the urine (qualitative reactions) with the
Combina kit, Germany, and by urinary sediment.
Hematological and biochemical studies were performed
after 10 and 30 days post the day of the experiment. At that
time animals were weighed. At the end of the experiment
some animals have been decapitated and the material has
been taken for pathomorphological studies according to the
list approved by Pharmacological Committee of the
Republic of Uzbekistan[5], The rest of animals has been left
to study the recovery process.
The general direction of changes in the coagulation
process under the influence of medication has been assessed
by the records of thromboelastograms performed on the
thromboelastograph Tromb-2.
The “open field” test is a study of behavior due to the
animal being in an unfamiliar open space, which it cannot
leave. The animal is put in the central square, and the latent
period of exit from it is recorder. The number of squares into
which the animal entered is registered (horizontal motion
activity - HMA), as well as the number of rises on the hind
limbs (“upright posture” - VMA), the number of holes the
animal snifled(exploratory activity - EA), the number of
washes (number of grooming acts -NGA) and defecations
(number of bolus - NB). Based on these parameters, motion
activity, orientative-motion activity, anxiety and autonomic
reactions are evaluated. The above parameters are fixed for
4 minutes from the moment of placing the animal
7
.
The conjunctival sample is a very sensitive test and,
in some cases, even discloses the reaction of animals to an
allergen at mild allergization and negative skin tests.
The trials have been performed on 12 rabbits, weighing 2,0-
2,5 kg, which have been instilled with 0,1 ml of 0,5 and 5%
medication solution in the left eye, and with 0,1 ml of
distilled water in the right eye (a controlled one). The
reaction has been recorded in 15 minutes (quick reaction)
and in 24-48 horns (delayed hypersensitivity) and evaluated
according to the following scale (in points). Besides, the
level of hyperemia, swelling, and lacrimation have
beenregistered
8
.
The local-irritant action of “Fitin-S” has been
studied on 12 rats weighed 150±10 grams. The animals have
had their hair clipped on both sides of the spinal column (4
areas) 2x2 cm in size.0,5 ml of Fitin-S in 0,5 and 5%
solutions have been applied on two clipped areas on the left
side of the back. The medication has been applied for 10
days. The controlled areas were the right clipped areas to
which distilled water has been applied in the same volume.
The monitoring has been carried out for 14 days. The skin
reaction has been recorded daily according to the skin
sample scale in points
8
.
Statistical processing of the obtained data has been
carried out with the definition of the Student’s criterion
using statistical programs WindowsExcel 2010.
All experimental animals have been kept in the
same conditions and on a common diet with free access to
water and food.
Results
The general action and “acute” toxicity of “Fitin-S”
have been determined in mice at a single intravenous,
intramuscular and subcutaneous administration. Each dose
of the substance has been studied in 6 animals. The
monitoring has been conducted for 14 days.
The medication has been administered to mice
intravenously at doses of 200,300,400, 500 and 600 mg/ kg
in the form of a 0.5% solution, intramuscularly and
subcutaneously at doses of 100-500-1000-2500-2700- 2800-
2900-3000-3300-3600 mg/kg as a 5% solution.
When studying the acute toxicity of the medication
“Fitin-S” with intravenous administration, a 5% solution of
the medication has been diluted with 0.9% sodium chloride
in a ratio of 1: 1; 1: 5 and 1:10. The obtained solutions of the
“Fitin-S” medication have been administered intravenously
at doses from 100 to 400 mg/kg in a volume not exceeding
0.5 ml per animal weight
3
. As can be seen from the data in
table 1, after
Medico-legal Update, October-December 2020, Vol. 20, No. 4
1219
administration of the medication at doses causing toxic
phenomena, convulsions and death of some mice have been
observed within 72 hours. The average lethal dose (LD50)
of intravenous administration of the “Fitin-S” medication to
mice has amounted to 347 (320 +- 460) mg/kg.
The results of studies with subcutaneous and
intramuscular administration have shown that “Fitin-S” at
doses of 100-1500 mg/kg does not cause changes in the
general condition of the animals. When the medication is
administered at doses of 1800-2500 mg/ kg, after 30-60
minutes there has been a decrease in the motion activity of
animals, while the response of mice to external stimuli has
remained unchanged, and no lethal outcome has been
observed. With the introduction of the medication at doses
of 2800-3600 mg/kg after 60-120 minutes, anxiety of mice,
shortness of breath, short-term convulsions and the death of
some animals during the day has been registered.
Table 1: The results of indicators of “acute” toxicity during parenteral administration of the medication “Fitin-S”
to mice
Animal species, type
of administration
Sex
Dose
mg/kg
Number of animals
in a group/number of
animals died
LD
10
-m+m
mg/kg
LD
16
-m+m
mg/kg
LD
50
-m+m
mg/kg
LD
S4
-m+m
mg/kg
Mice, intravenous
Males
150
6/0
233 -4,8 +6,0
294 -6,0 +7,6
347
-9,3
+13,1
460
-9,7
+12,3
200
6/2
250
6/4
300
6/5
400
6/6
Mice, intramuscular
Males
1800
6/0
2000
-380
+450
2266 -300 +330 2560 -350 +380 2893 -500 +550
2000
6/0
2200
6/2
2500
6/3
2800
6/5
3000
6/6
Mice, subcutaneous
2200
6/0
2240 -300 +330
2566
-320
+420
2900 -340
+380
3277
-440
+480
2500
6/1
2900
6/3
3000
6/4
3300
6/6
3600
6/6
Discussion
Thus, the study of the acute toxicity of “Fitin-S” has
shown that the medication is referred to class IV of low toxic
compounds. The LD
50
during intravenous, intramuscular
and subcutaneous administration to mice has amounted to
347(320+460) mg/kg respectively, 2560 (2266+2893)
mg/kg and 2900(2566-3277) mg/kg.
When choosing the doses for the study of chronic
toxicity of pharmacological substances their accumulative
effect should be taken into account. That’s why before
conducting experiments on chronic toxicity we have been
studying the accumulation via subchronic toxicity according
to the method of Lim and coauthors which allows to evaluate
not only accumulative properties but also addiction, and we
have determined
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Medico-legal Update, October-December 2020, Vol. 20, No. 4
the
accumulation index, the ratio of LD50 with a single
administration to LD50 with a multiple administration.
The studies conducted have shown that the
accumulation index K
K
has amounted toK
K
=LD
50n
/ LD
50
2944/2560=1,15. In other words, the medication “Fitin-S”
does not have the accumulative effect, the accumulation rate
is 1,15.
The local-irritant effect of the medication has been
studied on rats and rabbits. The results of monitoring have
shown that “Fitin-S” in 5 and 10% concentrations does not
cause even the slight reddening of the conjunctiva of the
rabbit eye in either 15 minutes or 24 or 48 hours. Application
of the medication for 10 days on the skin of rats also does not
cause irritation, redness, swelling or other visible changes on
the skin and the effect of “Fitin-S” is estimated at 0 points.
According to this a conclusion that the medication
“Fitin-S” in 5 and 10% concentrations does not have the
irritant effect on the conjunctiva of rabbits and on the skin of
rats can be made.
The
results
obtained
at
daily
intramuscular
administration of “Fitin-S” to rats and rabbits are represented
in Tables 3-4. As can be seen from the data given in tables 3-
4, in 10 and 30 administration at all the studied doses there
has been no significant deviations in the composition of the
peripheral blood of rats and rabbits in the hemoglobin
content of erythrocytes, in the number of blood cells
themselves, their precursors (reticulocytes), and leucocytes.
Leukocyte counts in rats and rabbits are within the
physiological norm.
Based on this, it can be concluded that “Fitin-S” at the
studied doses with intramuscular repeated administration
does not have a toxic effect on the quantitative composition
and morphology of the peripheral blood of experimental
animals.
Table 2: Indicators of peripheral blood of rats at daily intramuscular administration of the medication “Fitin-S”
(M±m; n=10)
Indicators
Control
10 days, dosemg/kg
30 days, dosemg/kg
20
40
100
20
40
100
Hemoglobin, gr/dl
14,7±0,3
15,8±0,7
15,9il,0
15,0±l,0
14,8i0,2
14,0i0,l
13,3i0,2
Erythrocytes, 10
12
/l
5,2±0,5
5,8i0,5
5,0i0,6
5,8i0,4
5,H0,3
4,8i0,2
5,0i0,6
Reticulocytes, %
4,2±0,2
5,4i0,3
5,8i0,3
3,9i0,3
6,5i0,4
6,5i0,5
4,3i0,4
Thrombocytes, 10
9
/l
675±11,5
830il2,5
600±ll,0
550il0,5
675il2
725il4
725il4
Leucocytes, 10
9
/l
10,2±0,8
ll,6±0,8
12,2±0,9
13,3i0,7
8,4i0,6
9,1±1,2
ll,9±0,7
Neutrophils:
Bands, %
l,8±0,2
2,0i0,2
2,0i0,2
2,0i0,2
2,0i0,2
2,0i0,2
2,0i0,2
Segmentonuclear, %
25,0±l,0
27,4i0,2
27,2i0,8
27,5il,0
28,5il,5
28,7il,3
28,5il,5
Eosinophils, %
2,0i0,2
2,0i0,2
2,0i0,2
2,0i0,2
2,0i0,3
2,0i0,2
2,0i0,2
Monocytes, %
3,0i0,3
2,6i0,2
2,9i0,3
3,0i0,3
3,0i0,3
3,0i0,3
3,0i0,3
Lymphocytes, %
64,2±1,8
66,0i3,4
66,8i3,2
65,5i3,5
65,Oil,6
64,3i2,7
64,5il,5
*P < 0,05 to control
Medico-legal Update, October-December 2020, Vol. 20, No. 4
1221
P > 0,05 to control
Table 3: Indicators of peripheral blood of rabbits at daily intramuscular administration of the medication “Fitin-
S” (M±m; n=4)
Indicators
Control
10 days, dosemg/kg
30 days, dosemg/kg
8
16
40
8
16
40
Hemoglobin, r/dl
9,6±0,3
9,9i0,7
12,0±0,5
H,2i0,l
9,2i0,7
8,4i0,5
8,4±0,l
Erythrocytes, 10
12
/1
4,5±0,5
4,HO,5
5,0i0,6
4,7i0,8
4,3i0,5
4,4i0,6
4,0i0,8
Reticulocytes, %
3,5±0,4
4,0i0,6
3,2i0,5
5,5i0,5
4,4i0,6
3,2i0,2
4,0i0,3
Thrombocytes, 10
9
/l
350±ll,5
250i6,5
225il3,0
400i8,5
250i6,5
325il3,0
400i8,5
Leucocytes, 10
9
/l
8,0±0,8
8,0i0,6
8,25i0,9
5,4i0,4*
9,4i0,6
9,0i0,9
8,4i0,4
Neutrophils:
Bands, %
l,8±0,2
2,2i0,2
l,9±0,2
2,0i0,2
l,8i0,2
2,0i0,2
l,9±0,2
segmentonuclear, %
25,0±l,0
27,4i0,2
27,2i0,8
27,5±l,0
27,0i0,2
27,2i0,8
27,5±l,0
eosinophils, %
2,0i0,2
2,0i0,2
2,0i0,2
2,0i0,2
2,0i0,2
2,0i0,2
2,0i0,2
basophils, %
l,0±0,l
0,8i0,6
l,li0,l
l,0±0,l
l,2i0,l
l,li0,l
l,0±0,l
monocytes, %
3,0i0,3
2,6i0,2
2,9i0,3
2,8i0,3
2,6i0,2
2,8i0,3
2,9i0,3
lymphocytes, %
63,2±1,8
65,0i3,4
65,8i3,2
64,7i3,5
65,0i3,4
65,8i3,2
64,5i3,5
*P < 0,05 to control
Diuresis after 10 days and 30 days of the experiment
under conditions of water load did not differ significantly in
the experimental and control rats. In rats, the state of
nitrogen metabolism, the criterion of which is the urea
content in the blood serum, did not deviate from the control
after 10 days and 1 month of the experiment. In the urine
during the entire experiment, both in the experimental
groups and in the control group, no changes in the studied
urine parameters have been detected. The acidity of urine in
all the studied groups has not been changing during the
entire experiment and its pH was 6.0-7.0. No pathological
changes in urine sediment have been detected.
The results obtained have led to the conclusion that the
medication “Fitin-S” with chronic administration does not
have a toxic effect on renal function in the studied doses.
The liver function in experiments on rabbits and rats has
been studied with the use of the most sensitive tests. The
levels of total protein, glucose, and enzyme activity (AsAT,
A1AT) in the blood serum of all experimental rats after 10
and 30 days of the experiment have not differed from the
control (Table 4.5).
Thus, the medication “Fitin-S” at intramuscular daily
administration at the studied doses does not have a toxic
effect on liver function.
Table 4: Some indicators of liver function of rats at intramuscular administration of the medication “Fitin-S” in a
chronic trial (M±m; n=5)
Study timeline
Dose mg/kg
Total protein,
gr/1
AlAt, mmol/l
AsAt, mmol/l
Glucosemmol/1
Urea mmol/l
10 days
Control
84i5,3
0,26i0,03
0,35i0,04
5, HO,4
4,4i0,42
“Fitin-S” 20
78i5,2
0,28i0,02
0,35i0,03
5,62i0,4
4,6i0,4
40
82i5,5
0,22i0,02
0,27i0,02
5,9i0,4
4,4i0,42
100
82i5,2
0,2 HO,02
0,39i0,04
5,9i0,4
4,6i0,4
30 days
Control
72i5,6
0,35i0,03
0,40i0,03
5,6i0,4
4,2i0,4
“Fitin-S” 20
86i5,6
0,3i0,02
0,36i0,03
5,5i0,4
4,3i0,4
40
76i6,0
0,28i0,02
0,36i0,02
5,6i0,4
4,5i0,42
100
76i6,0
0,28i0,02
0,4i0,03
5,5i0,4
4,6i0,4
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Medico-legal Update, October-December 2020, Vol. 20, No. 4
*P < 0,05 to control
Table 5: Some indicators of liver function of rabbits at intramuscular administration of the medication “Fitin-S”
in a chronic trial (M±m; n=5)
Study timeline
Dose mg/kg
Total protein,
gr/1
AlAt, mmol/l
AsAt, mmol/l
Glucosemmol/1
Urea mmol/l
10 days
Control
84±5,3
0,16±0,03
0,25±0,04
2,l±0,4
4,4±0,42
“Fitin-S”-8
68±5,2
0,18±0,02
0,22±0,03
0,62±0,4
4,6±0,4
16
82±5,5
0,18±0,02
0,27±0,02
l,9±0,4
4,4±0,42
40
82±5,2
0,21±0,02
0,3±0,04
2,9±0,4
4,6±0,4
30 days
Control
72±5,6
0,25±0,03
0,32±0,03
5,6±0,4
4,2±0,4
“Fitin-S”-8
86±5,6
0,3±0,02
042±0,03
5,5±0,4
4,3±0,4
16
76±6,0
0,26±0,02
0,36±0,02
5,6±0,4
4,5±0,42
40
76±6,0
0,28±0,02
0,34±0,03
5,5±0,4
4,6±0,4
P > 0,05 to control
As the medication “Fitin-S” is supposed to be applied
parenterally, it was important to find out how the process of
blood coagulation changes at multiple application. The
experiments have been held on 12 chinchilla rabbits, males
and females weighing 2,0±0,2 kg. The blood has been
studied in 10 and 30 days post the administration of “Fitin-
S” at doses of 8,0, 16,0 and 40,0 mg/kg.
The results of the impact of the medication “Fitin-S” on
the process of blood coagulation are represented in table 6.
Thromboelastogram data indicate that after 10 and 30
administrations, “Fitin-S” does not affect the blood
coagulation process at a therapeutic dose of 8 mg/kg. With
an increase in the therapeutic dose by 2 and 5 times (16 and
40 mg/kg), the blood coagulation process changes, which is
expressed in an increase in the indicator K, the time of clot
formation, which depends on the concentration of thrombin
formed and the amount of fibrinogen; and a decrease in MA,
a decrease in the elasticity of the clot (E) and the index of
hypercoagulation Ci, which indicates a slight decrease in the
process of blood coagulation.
Table 6: The dynamics of changes in blood coagulation of rabbits with chronic administration of the medication
“Fitin-S” M±m; n=4)
TEG Indicators
Control
In 10 days/dosemg/kg
In 30 days/dose mg/kg
8,0
16,0
40,0
8,0
16,0
40,0
R, mm
50±2,l
40±2,0
45±l,0
50±l,0
40±2,0
45±l,0
50±l,0
K, mm
15±l,0
15±1,1
25±l,0
30±0,6*
15±1,1
25±l,0*
30±0,6*
R+K, mm
65±3,5
55±3,0
70±3,0
80±6,6
55±3,0
70±3,0
80±6,6
R/K
3,3±0,2
2,8±0,l
l,8±0,l
l,7±0,l*
2,8±0,l
l,8±0,l*
l,7±0,l*
MA, mm
69±1,5
66±l,0
64±l,0
62±1,4
66±l,0
64±l,0
62±1,4
t, mm
100±10
100±10
100±10
100±10
100±10
100±10
100±10
S, mm
115±11
115±10
125±11
130±12
115±10
125±11
130±12
T, mm
165±14
155±14
170±15
180±16
155±14
170±15
180±16
Ci (MA/R+K)
l,l±0,l
l,2±0,l
0,9±0,l
0,8±0,06
l,2±0,l
0,91±0,l
0,8±0,06
E, MAxlOO/lOO-MA
222±19
194±16
178±15
163±14
194±16
178±15,0
163±14
ITC (E/S)
l,9±0,l
l,7±0,l
l,4±0,l
l,26±0, *
l,7±0,l
l,4±0,l
l,26±0,l*
Medico-legal Update, October-December 2020, Vol. 20, No. 4
1223
Thus, the intramuscular administration of the
medication “Fitin-S” to rabbits at a therapeutic dose of 8
mg/kg does not cause anyeffect, andat doses of 16.0 and
40.0 mg/kg leads to a slight slowdown in blood coagulation
process throughout the entire period of administration (30
days).
The study of the effect of the medication “Fitin-S” on
the central nervous system and peripheral nervous system
in the open field test has shown that after 10 days of
administration of “Fitin-S” for 4 minutes, it has not changed
the horizontal and vertical motion activity of rats at doses
of 20, 40 and 100 mg/kg. After 30 days of the experiment,
a sedative effect has been observed, which was expressed
in a reduction in the number of intersections (horizontal
activity) and the number of postures (vertical activity) and
the number of “holes” (exploratory activity). Throughout
the entire experiment, "Fitin-S'lias not significantly affected
the vegetative reactions of the div (the number of
“boluses”, the number of washings).
Table 7: Impact of “Fitin-S” on behavioral reactions of animals in the “open field” test (M±m; n=6)
Medication, dose,
mg/kg
Latentperiod,
sec
Motion activity
Quantity
Horizontal, number
oflntersections
Vertical, number
ofpostures
Holes
Washes
Bolus
In 10 days
Control
l,0±0,l
46,3±3,4
5,3i0,4
9,7±l,0
l,0±0
l,0±0,l
“Fitin” C-20
0,5±002
38,0±2,6
5,5i0,4
8,0i0,4
l,3±0,l
l,8±0,15
40
0±0
43,0±2,4
5,5i0,4
10,8±0,8
l,0±0,l
l,3±0,ll
100
0,75±002
49,0±3,6
7,8i0,6
6,0i0,4*
0,3±0,01
0,4i0,04
In 30 days
“Fitin” C-20
2,2±0,2
14,0±l,0*
2,5i0,2*
2,5i0,2*
0,45i00,2
l,0±0,3
40
3,8±0,2
32,3i3,0
3,6i0,2*
4,8i0,4*
0,6i0,04
l,4i0,l
100
l,0±0,l
26,0i2,3*
2,0i0,2*
5,0i0,4
l,3±0,l
0,5i0,02
*P < 0,05 to control
The study of integral indicators has shown that the
dynamics of the div weight of the experimental animals at
the studied doses after 10 and 30 days of the experiment has
not differed from the control. Throughout the experiment
animals were active, neat, they ate food normally, drank
water, their hair was smooth, shiny. The behavior of
experimental rats has not differed from the behavior of
control groups of animals.
A macroscopic examination of sacrificed animals has
disclosed the correct location of the internal organs. There is
no free fluid in the pleural and abdominal cavity. Tissues of
the lungs, stomach and intestines are also of a normal color,
without signs of edema, hemorrhages and ulcerations.
Pancreas, kidneys and adrenal glands are unchanged.
At the end of the experiment, 1 cm
3
of organs has been
taken
from experimental animals
for histological
examination. This site has been fixed in 10% normal
formalin. Then the tissue has been embedded in paraffin and
sections of histological preparations 3-5 microns thick have
been prepared and stained with a mixture of hematoxylin +
eosin. The obtained preparations have been analyzed under
a Leica-1000 DM microscope (Germany), Юх eyepiece,
lOx and 40xlens
6
The results of histological studies have shown that
basically their histostructure is comparable with the
morphology of control groups of animals. In the stomach,
the mucous membrane is well preserved, glandular cells are
without any changes. In the small intestine its main
elementsare villi and crypts. They are well prominent, villi
are finger-shaped, the border of enterocytes is distinguished,
the stroma of the villi and the epithelial layer are moderately
saturated with lymphocytes, are distinctly distinguished in
the enterocytes. In the colon, the crypts are lined with
mucous cells, the lumens of the crypts are enlarged and filled
with mucoid secretion.
In the liver of experimental animals treated with the
medication, no significant changes have been found. In
experimental animals at the studied doses, the majority of
hepatocytes are in a state of hydropic edema with granular
inclusions, and moderately medium-droplet fat inclusions on
the periphery. In addition, small foci of cellular infiltration
have beendisclosed in the liver parenchyma in some areas.
1224
Medico-legal Update, October-December 2020, Vol. 20, No. 4
Basically, as usual, the boundaries of the lobules are not
distinct in the liver, the hepatocytes are polygonal in shape,
their cytoplasm is pink and contains a large, roundish one or
two nuclei. In stellate macrophages, small grains of
hemosiderin have also been found, as well as in the control.
The structure of the various departments of the nephron,
renal corpuscles, proximal and distal parts are without any
changes. The structure of the capsule and cellular elements
of all departments of the nephron indicates their functional
viability.
White and red pulp are seen in the spleen; the structure
of these elements does not differ significantly from that of
control animals. The study of the structural organization of
the myocardium is indistinguishable from indicators in the
control group, consists of rectangular cardiomyocytes, with
a weakly pronounced transverse striation.
The lungs have a characteristic pattern and are
represented by numerous dilated alveolar vesicles and
atelectasis sites. The interalveolar walls contain numerous
blood capillaries, the walls of the bronchi and blood vessels
are without any significant changes.
The histology of the thymus is represented by the
cortical and brain zones, with signs of age-related
involution. The cortical zone is represented by lymphoid
cells and trabecular interlayers. Also, the brain area is
represented by lymphoid follicles, without clear centers of
reproduction.
In this regard, it should be noted that the differences
found are a manifestation of the species or age
characteristics of animals.
Thus, the results of histological studies allow us to
conclude that “Fitin-S” at the studied doses is the optimal
medication, its long-term use does not cause noticeable
structural destruction or damage and can be recommended
for further clinical trials.
Conclusion
Pre-clinical study of the general toxicity of the
medication “Fitin-S” has shown that it is referred to class IV
of low toxic compounds. The medication “Fitin-S” does not
have anaccumulative and local-irritant effect. With repeated
intramuscular administration to rats and rabbits, it does not
affect the behavior and dynamics of animal weight, does not
have a toxic effect on the composition of peripheral blood,
kidney and liver function, and also on the pathomorphology
of animal organs and tissues. With repeated use, there is a
slight slowdown in the blood coagulation process in rabbits
at doses of 16 and 32 mg/kg throughout the study period.
After 1 month of the recovery period, all these indicators
have been changing within the physiological norm. All of
the above data allow us to conclude that the medication does
not have a toxic effect on the animal organism.
Ethical Clearance:
No ethical approval is needed.
Source of Funding:
Self
Conflict of Interest:
Nil
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