Доклиническое исследование общей токсичности Препарат «Фитин-С»

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Мирзаахмедова, К., Зияева, Ш., Юнусов, А., Каримова, Г., & Каримов, Р. (2020). Доклиническое исследование общей токсичности Препарат «Фитин-С». in Library, 20(1), 1217–1225. извлечено от https://inlibrary.uz/index.php/archive/article/view/17281
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Аннотация

Доклиническое изучение общей токсичности препарата «Фитин-С» показало, что он относится к IV классу малотоксичных соединений. Препарат «Фитин-С» не обладает накопительным и местно-раздражающим действием. При многократном внутримышечном введении крысам и кроликам не влияет на поведение и динамику роста животных, не оказывает токсического действия на состав периферической крови, функцию почек и печени, патоморфологию органов и тканей животных. . Отмечается замедление процесса свертывания крови у кроликов в дозах 16 и 32 мг/кг в течение всего периода исследования. Через 1 месяц восстановительного периода все показатели изменялись в пределах физиологической нормы. Все вышеперечисленные данные позволяют сделать вывод, что препарат не оказывает токсического действия на организм животных.

Похожие статьи


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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


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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


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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


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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*


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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.


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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

References

1.

M.L. B. Elements for qualitative evaluation of the

pharmacological effect [Element! kolichestvennoi
otsenki farmakologicheskogo effekta]. L. «Medgiz».
1963.

2.

T.A G. Toxicology of medications [Toksikologiya

lekarstvennykh sredstv]. Moscow. 2008;: p. 27-30.

3.

A.V. Stefanov. K. Pre-clinical study of medications

(guidelines)

[Doklinicheskoye

issledovaniye

lekarstvennykh

sredstv

(metodicheskiye

rekomendatsii)]. 2002;: p. 587.

4.

A.N. M. Guidelines for the pre-clinical study of the

general toxic effects ofmedications [Metodicheskiye

rekomendatsii

po

doklinicheskomu

izucheniyu

obshetoksicheskogo

deistviya

lekarstvennykh

sredstv]/Guidelines

for

pre-clinical

studies

of

medications. Parton. 2012;: p. 13-23.

5.

Instructions for preclinical

safety testing

of pharmacological agents

[Instrukcii po

doklinicheskomu

ispitaniyu

bezopasnosti

farmakologicheskikh sredstv]. Tashkent. 2000.

6.

others BVPa. Laboratory method for studying the

hemostasis

system

[Laboratomiye

metody

issledovaniya sistemy gemostaza]. Tomsk. 1980.

7.

Markel G.A. KRA. The method of complex

registration of behavioral and autonomic reactions in
rats during the open “field test” [Metod kompleksnoi
registratsii povedencheskikh I vegetativnykh reaktsiy u

kris pri provedenii testa otkritoye pole]. Zhum.vish.

nervn. deyat. 1976; 6(26): p. 1314-1318.

8.

Meditsina ebDSSaYLPM. Preparation of histological

sections.

(Microscopic

Technique:

Manual)

[Prigotovleniye

gistologicheskikh

srezov.

(Mikroslopicheskaya tekhnika: Rukovodstvo)]. 1996;

p. 432.

9.

Mirzaahmedova, K. T., Abdullaeva, S. K.,

Akhmadiev, E. E., & Ziyayeva, S. H. (2017). The
effect of immunomoduline and phytin compounds
on lipid peroxidation induced at toxic experimental


background image

Medico-legal Update, October-December 2020, Vol. 20, No. 4

1225

hepatitis.

In

International

Conference

on

Chemical, Biological and Health Sciences

(pp. 99-

106).

10.

Калдыбаева, А. О., and А. А. Абдусаматов.

"Влияние

корня

солодки,

цветков

бессмертника и мумиё на показатели
перекисного

окисления

липидов

при

хроническом

токсическом

гепатите."

Медицинский

журнал

Узбекистана

3 (2010): 99-101.

11.

Мирзаахмедова, К. Т., А. А. Абдусаматов, and

А. Н. Набиев. "Влияние иммуномодулина и
соединений

фитина

на

показатели

перекисного

окисления

липидов

при

экспериментальном

токсическом

гепатите."

Сибирский медицинский журнал

(Иркутск)

88.5 (2009): 122-124.

12.

Aminov,

S.

D.,

and

A.

A.

Vakhabov.

"Pharmacology of some coumarins isolated from
plant Haplophyllum."

Dokl. Acad. Nauk. UzSSR

8

(1985): 44-45.

13.

Аминов, С. Д., and К. Т. Мирзаахмедова.

"Изучение

гастропротекторных

свойств

глицитрината

на

экспериментальных

животных."

Педиатрия

1-2 (2013): 118.

Библиографические ссылки

M.L. B. Elements for qualitative evaluation of the pharmacological effect [Element! kolichestvennoi otsenki farmakologicheskogo effekta]. L. «Medgiz». 1963.

T.A G. Toxicology of medications [Toksikologiya lekarstvennykh sredstv]. Moscow. 2008;: p. 27-30.

A.V. Stefanov. K. Pre-clinical study of medications (guidelines) [Doklinicheskoye issledovaniye lekarstvennykh sredstv (metodicheskiye rekomendatsii)]. 2002;: p. 587.

A.N. M. Guidelines for the pre-clinical study of the general toxic effects ofmedications [Metodicheskiye rekomendatsii po doklinicheskomu izucheniyu obshetoksicheskogo deistviya lekarstvennykh sredstv]/Guidelines for pre-clinical studies of medications. Parton. 2012;: p. 13-23.

Instructions for preclinical safety testing of pharmacological agents [Instrukcii po doklinicheskomu ispitaniyu bezopasnosti farmakologicheskikh sredstv]. Tashkent. 2000.

others BVPa. Laboratory method for studying the hemostasis system [Laboratomiye metody issledovaniya sistemy gemostaza]. Tomsk. 1980.

Markel G.A. KRA. The method of complex registration of behavioral and autonomic reactions in rats during the open “field test” [Metod kompleksnoi registratsii povedencheskikh I vegetativnykh reaktsiy u kris pri provedenii testa otkritoye pole]. Zhum.vish. nervn. deyat. 1976; 6(26): p. 1314-1318.

Meditsina ebDSSaYLPM. Preparation of histological sections. (Microscopic Technique: Manual) [Prigotovleniye gistologicheskikh srezov. (Mikroslopicheskaya tekhnika: Rukovodstvo)]. 1996; p. 432.

Mirzaahmedova, K. T., Abdullaeva, S. K., Akhmadiev, E. E., & Ziyayeva, S. H. (2017). The effect of immunomoduline and phytin compounds on lipid peroxidation induced at toxic experimental hepatitis. In International Conference on Chemical, Biological and Health Sciences (pp. 99-106).

Калдыбаева, А. О., and А. А. Абдусаматов. "Влияние корня солодки, цветков бессмертника и мумиё на показатели перекисного окисления липидов при хроническом токсическом гепатите." Медицинский журнал Узбекистана 3 (2010): 99-101.

Мирзаахмедова, К. Т., А. А. Абдусаматов, and А. Н. Набиев. "Влияние иммуномодулина и соединений фитина на показатели перекисного окисления липидов при экспериментальном токсическом гепатите." Сибирский медицинский журнал (Иркутск) 88.5 (2009): 122-124.

Aminov, S. D., and A. A. Vakhabov. "Pharmacology of some coumarins isolated from plant Haplophyllum." Dokl. Acad. Nauk. UzSSR 8 (1985): 44-45.

Аминов, С. Д., and К. Т. Мирзаахмедова. "Изучение гастропротекторных свойств глицитрината на экспериментальных животных." Педиатрия 1-2 (2013): 118.

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