International Journal of Scientific Pediatrics
STUDYING THE PHARMACOKINETICS OF GENTAMYCIN IN RATS WITH
LYMPHOTROPIC PRETRACHEAL AND INTRAMUSCULAR INTRODUCTION
Mamatov B.Yu.
1
, Muminov B.E.
2
, Kuziev O.A.3, Abdullaev A.S.
4
, Sobirov M.S.
5
, Ismailov O.A.
6
, Ergashev Kh.M.
7
,
Khasanov Sh.N.
8
, Atahanova N.S.
9
1 Andijan State Medical Institute, Andijan, Uzbekistan. Candidate of medical sciences,
professor of Anesthesiology - Resuscitation, pediatric anesthesiology – resuscitation.
2 Andijan State Medical Institute, Andijan, Uzbekistan. Senior Lecturer Department of
Anesthesiology - Resuscitation, pediatric anesthesiology – resuscitation.
3 Andijan State Medical Institute, Andijan, Uzbekistan. Assistant Department of
Anesthesiology - Resuscitation, pediatric anesthesiology – resuscitation.
4 Andijan State Medical Institute, Andijan, Uzbekistan. Assistant Department of
Anesthesiology - Resuscitation, pediatric anesthesiology – resuscitation.
5 Andijan State Medical Institute, Andijan, Uzbekistan. Senior lecturer Department of
Anesthesiology - Resuscitation, pediatric anesthesiology – resuscitation.
6 Andijan State Medical Institute, Andijan, Uzbekistan. Assistant Department of
Anesthesiology - Resuscitation, pediatric anesthesiology – resuscitation.
7 Andijan State Medical Institute, Andijan, Uzbekistan. Assistant Department of
Anesthesiology - Resuscitation, pediatric anesthesiology – resuscitation.
8 Andijan State Medical Institute, Andijan, Uzbekistan. Assistant Department of
Anesthesiology - Resuscitation, pediatric anesthesiology – resuscitation.
9 Andijan State Medical Institute, Andijan, Uzbekistan. Assistant at the department of
Propaedeutic of Internal Diseases.
Academic Editor:
Arzikulov A.
Professor, Andijan State Medical
Institute
Received:
15 may 2022
Accepted:
28 May 2022
Published:
31 May 2022
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authors. Licensee IJSP, Andijan,
Uzbekistan. This article is an open
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Аннотация.
The article is devoted to an experimental study of the pharmacokinetics
of gentamicin in rats with lymphotropic pretracheal and intramuscular routes of
administration. Data on the concentration of the antibiotic in the blood and tissues of
the respiratory organs of rats in dynamics are given, it is shown that the method of
lymphotropic pretracheal administration provides higher concentrations of the antibiotic
in comparison with the intramuscular method. The high therapeutic efficiency of this
method can be achieved due to the presence of connections between the lymphatic
system, as well as organs and tissues. This circumstance allows you to deliver the drug
to the affected organ directly. In addition, in the case of lymphotropic therapy in the focus,
the optimal concentration of the drug is maintained for 24 hours. So there is no need for
frequent administration of the drug (for example, with antibiotic therapy), you can reduce
the course and daily doses of drugs. The drug, which is administered lymphotropically, is
not able to have a toxic effect on the intestines, kidneys, liver, since only a tiny part of the
drug enters the bloodstream.
Key words:
gentamicin, method of lymphotropic pretracheal administration,
intramuscular administration, concentration, focus, optimal concentration.
Article
OPEN ACCESS
published: 31 May 2022
doi.org/10.56121/2181-2926-2022-1-30-40
May 2022 / Issue 01 / Article 04
30
IJSP
Introduction.
The issues of prophylactic antibiotic therapy remain
very relevant and far unresolved. Some authors point out the need for
prophylactic antibiotic therapy when the risk of developing a purulent-septic
process is high, S.M. Navashin and I.P. Fomina believe that prophylactic
antibiotic therapy in surgery is indicated in the following cases: operations
in obviously infected areas (on the gastrointestinal tract), general infection
before surgery, weakening of the div’s defenses.
The success of antibiotic therapy depends not only on the activity of
the drugs and the sensitivity of microorganisms to them, but also on the
duration of the preservation of the therapeutic concentration of antibiotics
in the affected tissue and on the path of infection. First of all, this applies
to the lymphatic system [1, 4, 9, 5]. Despite this, in the literature, there are
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quite rare reports on the content of antibiotics in the lymphatic system of
the organs of the chest cavity, and very few researchers pay attention to
this issue [2, 6, 8]. This state of affairs has its own explanation, connected
with the difficulties of obtaining lymph for research, not only in patients, but
also in experimental animals.
Recommended in clinical practice schemes of intramuscular and
intravenous antibiotic therapy [3,7] do not allow to create a sufficient and
long-term concentration of the drug in the lymphatic system of the lungs
[2, 10].
The question of the spread of antibiotics in lymph nodes regional
in relation to the affected organ with traditional methods of administering
antibiotics. There are only a few reports in the literature on this subject,
although this issue is of fundamental importance due to the fact that it is
the regional lymph nodes that are the first «biological filter» on the path
of the spread of microbes and, under certain conditions, can themselves
become a source of infection.
Among other reasons for the ineffectiveness of antibiotic therapy,
the authors point to the erroneously chosen route of administration of
drugs into the div, as well as their overdose, which leads to a general
toxic and negative effect of antibiotics on individual organs and systems.
Due to the decrease in the activity of antibiotics to achieve a therapeutic
effect in recent years, many clinicians are forced to increase the doses
of administered drugs to a critical level, which leads to an increase in
undesirable side effects, such as allergic reactions, ranging from skin
lesions to anaphylactic shock and occurring from 0,3 to 4,8%. The defeat
of the central nervous system is noted in 1-10% of cases. Often there is
toxic damage to internal organs: liver, kidneys, gastrointestinal tract - in
2.4% of cases, manifestations of «dysbacteriosis» and «superinfection»
[8, 2]. The influence of large doses of antibiotics on the div’s defenses
by inhibiting immunogenesis has been proven [5].
Thus, summarizing the above literature data, we can assume that
among the reasons for the insufficiently high efficiency of antibiotic therapy,
the circumstance is important that with traditional methods of administering
antibiotics, it is often not possible to achieve a therapeutic effect due to the
limitation of the spread of drugs in the affected tissues and biological media
of the div, in particular in the lymph. This causes the lack of sufficient
contact between the antimicrobial drug and microorganisms, which leads
to the emergence of antibiotic-resistant microbes and an increase in
antigenic irritation from bacteria. It is also important that with the above
methods of antibiotic therapy, the drug is quickly excreted from the div,
which obliges frequent injections of antibiotics to maintain their therapeutic
concentration in the blood, tissues and biological media. All this leads to the
need to increase the doses of administered antibiotics and the frequency
of injections, increasing the risk of side effects of antibiotic therapy.
According to a number of authors [5, 2], possible directions in the
search for ways to increase the effectiveness of antibiotic therapy are to
find ways to increase the concentration of antibiotics in affected tissues
and environments, as well as to develop new routes of administration of
drugs (endolymphatic, lymphotropic, regional, et c.), taking into account
the laws spread of infection in the div.
The study of the lymphatic tracts of the lungs, liver, heart, appendix,
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gallbladder in pathological conditions, carried out by the authors [3, 10],
made it possible to identify compensatory changes in the lymphatic system
common to all these organs, despite the fact that these organs are different,
with point of view of blood circulation, systems. With inflammation of these
organs, an active restructuring of the structure of the lymphatic tract
occurs, the degree of which depends on the stage, form and activity of the
pathological process. In the initial period of inflammation, there is a general,
relatively uniform expansion of the lymphatic capillaries and vessels of all
orders with a quantitative and qualitative increase in outgrowths on their
walls. This leads to an increase in the capacity of the lymphatic system
and the creation of conditions for the removal of an increased volume of
lymph. Along with the functional reaction, a regenerative-compensatory
morphological restructuring of the lymphatic system is always found,
which manifests itself in the neoplasm of lymphatic vessels and capillaries,
intercalary lymph nodes. New collector paths of lymph outflow appear in the
form of separate, independent single, atypically located lymphatic vessels.
In this case, the physiological flow of lymph from the organs to the thoracic
duct may change and lymph flow occurs. The formation of lymphatic flow
is important for understanding the mechanisms of endolymphatic antibiotic
therapy. There are new collector ways of outflow of lymph in the form of
separate, independent single, atypically located lymphatic vessels. In this
case, the physiological flow of lymph from the organs to the thoracic duct
may change and lymph flow occurs. The formation of lymphatic flow is
important for understanding the mechanisms of endolymphatic antibiotic
therapy. There are new collector ways of outflow of lymph in the form of
separate, independent single, atypically located lymphatic vessels. In this
case, the physiological flow of lymph from the organs to the thoracic duct
may change and lymph flow occurs. The formation of lymphatic flow is
important for understanding the mechanisms of endolymphatic antibiotic
therapy.
Collector lymphatic vessels play an important role in limiting
inflammation. And for them, as well as for lymphatic capillaries, structural
and functional changes are characteristic. Already in the early stages of
inflammation, swelling of the endothelium, the opening of interendothelial
connections, and the expansion of the lumen of the lymphatic vessels
due to their overflow with lymph are noted. These phenomena contribute
to the alteration of the vascular endothelium in the form of a catarrhal
process, which leads to activeendothelial cells, which acquire the functions
of macrophages. A number of authors indicate that infection directly into
the lymphatic system contributes to the immune response through the
functional activation of lymphoid tissue [1, 3, 7]. However, under conditions
of increased virulence of microbes, lymphatic vessels are involved in the
inflammatory process with the formation of lymphangiitis. Thus, in the
lymphatic capillaries and collector lymphatic vessels during inflammation,
there is a similar functional response and compensatory morphological
restructuring.
The presentation of the function of the lymphatic system of the lungs
in the dynamics of the inflammatory process would not be complete if we
did not dwell on the role of the lymph nodes. Lymph nodes are a biological
filter. They are located in several orders along the collector lymphatic
vessels, which ensures the obligatory passage of lymph through them,
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and only then the lymph flows into the thoracic duct and bloodstream.
The main part of the lymph from the right and left lungs flows into
the paratracheal lymph nodes, which serve as a rather powerful collector
that receives lymph from the lungs. That is why these lymph nodes
should serve as the place of application of lymphotropic regional antibiotic
therapy. In addition, the paratracheal lymph nodes are the most accessible
for regional lymphatic therapy. Thus, the lymphatic system of the lungs is
important in the development and course of the inflammatory response.
The spread of infection along the lymphatic channel dictates the need
for the introduction of antibiotics precisely inlymphatic system in order to
prevent and treat bronchopulmonary bacterial complications.
From the point of view of rationality and economy in choosing the
place of application of microcirculation correctors, antibacterial agents and
in order to create a more pronounced regional tropism, S.U. Dzhumabaev
et al. a method of pretracheal lymphatic therapy was developed, in which
the drug is administered by puncture or catheterization of the pretracheal
tissue on the anterior surface of the neck above the jugular notch. In
experimental studies on the introduction of the dye by this method,
the authors showed that it easily spreads down the fiber, washing the
main groups of lymph nodes that are around the trachea and bronchi.
They also successfully use the method in the prevention and treatment
of postoperative pulmonary complications associated with circulatory
disorders (edema) and inflammation.
However, there are also a number of unresolved issues, in particular,
the pharmacokinetics of antibiotics with lymphotropic administration by
this method has not been studied, there is no data on their distribution in
blood, regional lymph nodes and tissues. The optimal place of the method
of pretracheal lymphotropic administration of antibiotics among the existing
methods has not been finally determined.
Мethods.
Experimental studies consisted of two stages. The first
stage consisted in solving a particular experimental problem. It consisted
of a single administration of gentamicin to outbred white rats by one of
the studied methods - pretracheally and intramuscularly - followed by
a thoracotomy undertaken to remove the organs of the chest cavity,
paratracheal lymph nodes and puncture the femoral vein for blood
sampling. The second stage consisted in determining the concentration
of gentamicin in the blood and removing tissues for the preparation of a
homogenate.
Gentamicin was administered once to rats at a dose of 30 mg/kg.
The experiments were carried out on 50 outbred white rats of both sexes
weighing 190-230 g. Two series of experiments were carried out.
In the first series of experiments, 25 rats were pretracheally injected
with lidase at a dose of 0.1 U/kg to create conditions for lymphotropism. 3-5
minutes after the injection of lidase, the needle was pulled up by 0.5 cm and
gentamicin was administered once at a dose of 30 mg/kg. In the second
series of experiments, gentamicin was administered intramuscularly to 25
rats once at the same dose along the anterolateral surface of the hind
paw. This group of animals served as a control, with the aim of studying
the distribution of gentamicin in the traditional intramuscular method of
administration.
To determine the concentration of gentamicin, blood sampling in a
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volume of 1-5 ml and pieces of organ tissues were performed 1, 3, 5, 8, 24
hours after the administration of the antibiotic.
At the specified time, 25 animals were slaughtered in both groups,
from which tissues of the paratracheal lymph nodes, trachea with bronchi,
lungs, pleura and blood were taken.
The concentration of gentamicin in the blood and supernatant of the
tissue homogenate was determined by agar diffusion.
Results.
Analysis of the dynamics of the concentration of gentamicin
with a single pretracheal lymphotropic injection at a dose of 30 mg/kg of
div weight showed that the maximum concentration of the antibiotic in
the blood and tissues of the respiratory organs is observed after 1 hour,
as with the intramuscular route of administration, regardless of prior
lymphostimulation with lidase. So, in the blood serum, the concentration
of the antibiotic was 40,9 ± 0,49 mcg /ml after 1 hour, and after 3 hours
it remained at the level of 25,9±0,49mkg/ml. By the end of the day, the
antibiotic concentration decreased to a level equal to 0,036±0,002 mkg /
ml. The concentration area under the curve in this case was 15,2 cm² (Fig.
1.).
Fig.1. Dynamics of the concentration of gentamicin in the blood
after a single pretracheal lymphotropic and intramuscular
administration
In the tissues of the paratracheal lymph nodes, the maximum
concentration of gentamicin after an hour was 90,5±12,4 mkg/mg, after 3
hours there is a decrease in the concentration of the antibiotic, amounting
to 56,3±8,4 mkg/mg, then during the day the concentration decreases
evenly, after 24 hours it was 3,6±0,19 mkg/mg. The total area under the
curve with pretracheal lymphotropic injection was 41 cm² (Fig. 2.).
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Fig.2. Dynamics of gentamicin concentration in paratracheal lymph
nodes after a single pretracheal lymphotropic and intramuscular
administration.
In the tissues of the trachea and bronchi, the maximum concentration
of gentamicin after 1 hour is 80,3±10,5 mkg/mg, after 3 hours the content
of gentamicin decreases to 72,4±9,3 mkg/mg. During the day, there is a
uniform decrease in the level of the antibiotic and after 24 hours its content
remains at the subtherapeutic level and is equal to 4,9±0,3 mkg/mg. In
graphical analysis, the concentration area under the curve was 47,0 cm²
(Fig. 3.).
Fig.3. Dynamics of gentamicin concentration in the trachea and
bronchi after a single pretracheal lymphotropic and intramuscular
administration.
In the lungs with pretracheal lymphotropic administration, the content
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of gentamicin after 1 hour was 102,4±13,3 mkg / mg, decreasing after 3
hours to a level of 53,3±3,3 mkg /mg. During the day, its concentration
decreases smoothly, after 24 hours it was equal to 9,8±0,44 mkg /mg.
With a graphical representation, the area under the curve was 50,9 cm²
(Fig. 4.).
Fig.4. Changes in the concentration of gentamicin in the lung
tissue after a single pretracheal lymphotropic and intramuscular
administration.
In the pleural tissue, the antibiotic content after 1 hour was 38,6±3,2
mkg / mg, after 3 hours its concentration decreased to 18,9±0,9 mkg / mg,
and during the day the concentration of gentamicin with both methods of
administration was approximately the same values, equaling 0,58 mkg /
mg after 24 hours with pretracheal administration. In the graphical analysis
of the obtained data, the area under the curve was 13,2 cm² (Fig. 5.).
From the above data of experimental studies, it follows that higher
concentrations of the drug in the above biological substrates are provided
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with the lymphotropic pretracheal route of administration of gentamicin
compared with the intramuscular route of administration. To illustrate
this, we present a table of comparative dynamics of the concentration
of gentamicin in the studied substrates after a single intramuscular and
pretracheal administration in the same dose.
T
able 1
The concentration of gentamicin in blood serum (mcg/ml) and
tissues (mcg/mg) with a single intramuscular and lymphotropic
pretracheal injection at a dose of 30 mg/kg
Note: v-intramuscular, l-lymphotropic method: *-significantly
different values for the compared routes of administration (P<0.05).
For a more complete and visual representation of the kinetics of the
antibiotic in the blood and tissues, it is advisable to conduct a comparative
analysis of the concentrations obtained with the lymphotropic pretracheal
and intramuscular routes of administration. An analysis of the distribution
of gentamicin in the blood of white rats with various routes of administration
shows that the intramuscular route of administration (traditional) creates
the maximum peak concentration, among other indicators, during the
first hour, but by 3 hours there is a decrease in the concentration of the
antibiotic by more than 2 times from the initial value . And after 5 hours,
the intramuscular method does not provide a therapeutic concentration of
the drug in the blood. If the peak concentration of gentamicin in the studied
media after 1 hour is conditionally taken as 100%, then its concentration
in the blood by the 3rd hour decreased by 57,3%, and by the 5th hour,
amounting to 0,17%, decreased by more than 99%. With the lymphotropic
method of administration, the highest concentration also falls on the period
of the first hour, which indicates a relatively rapid absorption of the drug in
a sufficiently high concentration into the blood, despite the swelling of the
pretracheal tissue created by the use of lidase as a lymphatic drainage
stimulator. The dynamics of the concentration of gentamicin in the blood
after lymphotropic administration after 3 hours decreased by 37,4%, by 5
hours - by 93%, by 8 hours - by 99%. Data on the content of gentamicin
in the blood for both methods of administration after 24 hours are almost
identical. This dynamics indicates that after 5 hours the content of the
antibiotic with the lymphotropic method of administration remains at a
biological substrate
Time after administration (h)
1
3
5
8
24
Blood
in
238,2±18,2
101,8±7,8
0,41±0,06 0,23±0,02 0,019±0,004
l
40,9±0,49
25,6±0,49
2,9±0,1*
0,43±0,1
0,036±0,002
paratracheal
nodes
in
35,5±1,2
26,8±0,2
7,4±0,2
5,1±0,49
1,4±0,12
l
90,5±12,4*
56,3±8,4*
13,9±1,2* 6,7±1,02*
3,6±0,19*
trachea bronchi in
70,2±7,3
68,8±10,1
27,4±0,5
6,5±0,1
3,7±0,2
l
80,3±10,5*
72,4±9,3*
45,3±2,4* 14,6±0,7*
4,9±0,3
Lungs
in
93,3±3,2
41,8±2,3
33,5±0,6
12,8±1,2
5,2±0,46
l
102,4±13,3*
53,3±3,3*
48,9±0,5* 20,1±1,2*
9,8±0,44*
Pleura
in
23,9±0,49
6,2±0,49
1,8±0,19
1,7±0,22
0,48±0,05
l
38,6±3,2*
18,9±0,9*
1,5±0,12
1,4±0,1
0,58±0,05
International Journal of Scientific Pediatrics
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subtherapeutic level of 2,9-0,1 mkg /ml, which is 7 times higher than the
corresponding indicator obtained with intramuscular injection. In a graphical
representation, the concentration curve for lymphotropic administration is
smoother than for intramuscular administration. After 24 hours, with both
methods of administration, only “traces” of the antibiotic remain in the
blood, and with lymphotropic administration, it is approximately 2 times
higher than that with intramuscular administration.
And (&) Discussion.
Analyzing the data of experimental studies on
the content of gentamicin in the blood, it can be said that the pretracheal
lymphotropic method of antibiotic therapy using a single dose of 30 mg/
kg in rats provides, compared with intramuscular administration, although
not high, but more stable and long-term maintenance of blood saturation.
Comparison of the content of gentamicin in the paratracheal lymph
nodes with different methods of administration showed that the highest
concentration is created after 1 hour with lymphotropic pretracheal
administration, amounting to 90,5±12,4 mkg/mg versus 35,5±1,2 mkg /mg
intramuscularly. After 3 hours, the content of the drug with intramuscular
injection, amounting to 75% of the peak concentration, respectively, was
lower than the corresponding indicator for lymphotropic administration by
more than 2 times. In the future, during the day, this trend continues, there
is a sharp decrease in the content of the antibiotic when administered
intramuscularly. After 5 hours, it decreases by 3,5 times, and during this
period, with lymphotropic administration, the concentration of the drug is
1,8 times higher than that with intramuscular administration. The content
of the drug in the lymph nodes obtained with lymphotropic administration
after 8 hours is 1,3 times higher than that with intramuscular injection. After
24 hours, the lymphotropic concentration is 2,5 times higher than that with
intramuscular administration of gentamicin.
Thus, lymphotropic pretracheal administration of an antibiotic showed
that this method makes it possible to create in the paratracheal lymph
nodes relatively more and long-term concentrations of the drug, which
remain at the therapeutic and subtherapeutic levels during the day.
As for the pharmacokinetics of gentamicin in the respiratory organs,
in the tissues of the trachea and bronchi in the first hour after lymphotropic
administration, the concentration obtained is higher than with intramuscular
administration by 16,1 mkg/mg (18,7%). After 5 hours, the intramuscular
content of the drug decreases by more than 2,5 times, while it is also
more than 2 times lower than the corresponding content of gentamicin in
the trachea obtained with lymphotropic administration. In the future, the
concentrations obtained with intramuscular injection after 8 and 24 hours
are several orders of magnitude lower than those obtained with lymphotropic
administration. The graphical analysis also shows that the concentration
curve for the lymphotropic route of administration is smoother and more
uniform. The total area of concentration under the curve with lymphotropic
administration is 33,2% higher than that with intramuscular administration
of the antibiotic.
In lung tissues, the maximum concentration after 1 hour with
lymphotropic administration is higher than that with intramuscular
administration by 8,9%, the content of the antibiotic after 3, 5 and 8 hours
with lymphotropic administration is also higher than with intramuscular
administration, respectively, by 21,6, 31,5 and 42,7%. Further, after 24
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hours, the difference in the content of the antibiotic in the tissues of the
lungs with the compared methods of administration is significant. It is
almost 2 times higher with lymphotropic administration of the corresponding
concentration with intramuscular administration. It should also be noted that
after 24 hours, the content of the antibiotic with intramuscular administration
decreases by 18 times from the initial maximum concentration, and with
lymphotropic - by 14,4 times. The decrease in lymphotropic concentration
in the tissues of the lungs, as in other substrates, occurs more evenly and
smoothly than with intramuscular administration.
Analysis of the content of the antibiotic in the pleural tissue showed
that the values of the maximum concentrations are the lowest among
identical ones in other tissues, with the highest value equal to 38,8±3,3
mkg /mg, obtained with lymphotropic administration and 1,6 times higher
than identical with intramuscular administration. After 3 hours, the gap
between the corresponding indicators increases by 3 times. In subsequent
time periods, the indicators are approximately the same. After 24 hours,
only «traces» of the antibiotic in the pleura were determined with both
methods of administration. The total area under the curve with lymphotropic
administration is almost 45,9% more than that with the intramuscular
method.
When comparing the average daily content of gentamicin in tissues
in relation to the content in the blood, it turned out that the indicators
obtained with lymphotropic administration are also higher than those with
intramuscular administration.
Thus, the obtained experimental data proved that relatively high
and long-lasting concentrations in the paratracheal lymph nodes and
respiratory organs are achieved with the lymphotropic pretracheal route
of administration than with the intramuscular route, which undoubtedly
indicates the advantage of this method over the traditional one.
It should also be noted that with intramuscular administration, there
are sharp fluctuations in the content of the drug in the blood, which is not the
case with lymphotropic administration. The relatively low peak content of
gentamicin in the blood with lymphotropic administration, which exceeds the
average therapeutic concentrations, a slow decrease in the concentration
of the antibiotic in the blood can provide a general therapeutic effect. As
for the content of gentamicin in the paratracheal lymph nodes and tissues
of the respiratory organs in rats, high therapeutic concentrations are noted
here with a slow decrease during the day. It is also valuable that with a
single lymphotropic administration of the indicated dose of an antibiotic, its
long-term concentrations are retained in all tissues, which is an important
point in the prevention and treatment of bronchopulmonary complications.
The obtained dynamics of gentamicin concentrations in the studied
substrates allows us to resolve the issue of the multiplicity of lymphotropic
and intramuscular administration of the drug.
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