CC BY
29
© D. M. Boyko, M. G. Boyko, D. E. Nikolenko
UDC 616. 24-002-092. 9-08
D. M. Boyko, M. G. Boyko, D. E. Nikolenko
AN ANALYSIS OF THE DIFFERENT VARIANTS’ IMPACT OF ANTIINFLAMMATORY THERAPY ON THE COURSE INITIATED BY THE SIO (2) GRANULOMETOUS INFLAMMATION IN THE LUNGS OF RATS
The higher state educational institution of Ukraine «Ukrainian medical stomatologic Academy» (Poltava)
Work is a fragment of the scientific-research theme: «Clinico-functional and morphological characteristics of the course of diseases of the respiratory system (tuberculosis, sarcoidosis, disemination processes and CNLD) at different stages of treatment, rehabilitation and prevention», number of state registration 0110U008151.
Introduction. The problem of granulomatous diseases of respiratory organs for today is receiving increasing attention, in particular sarcoidosis [4], which belongs to a group of interstitial respiratory diseases of unknown etiology, and is characterized by the accumulation of activated T-cells and mononuclear phagocytes with the formation of epithelioid-cell granulomas without necrotizing in the affected organs [4, 21]. The majority of epidemiological and laboratory studies suggest that the disease reflects the immune inflammatory response to the etiological agent in genetically predisposed individuals [1, 4, 9]. A number of factors, which may cause the development of the granulomatous inflammation: bacteria, fungi and parasites; products of plants and animals (pollen, spores, proteins); metal compounds [1, 4, 6, 9]. There are also scientific evidence that the contact with the silicates may be associated with the emergence of sarcoidosis [8].
For experimental playback of granulomatous inflammation (GI) in order to verify the hypotheses and efficacy of medicines, determination of the characteristics of the course of disease, in the basis of the pathogenesis of which is inflammation with the formation of granulomas - use experimental model of GI, based on the introduction into the organism of laboratory animals of the suspension particles SiO(2) [19].
Taking into account the above-stated data for the modelling of granulomatous inflammation in laboratory animals, it was planned to use instillation in lungs of the suspension particles SiO(2). And for the correction of inflammatory changes in the lung tissue offered drugs with anti-inflammatory and antifibrous properties, namely: glucosamine sulfate [7, 13, 14], dexametha-sone [11, 15, 20, 23] and the combination of imunofani [12, 17, 22, 24] and hyaluronidase [10, 16, 18]. In our opinion this unusual choice of anti-inflammatory therapy will enhance our knowledge about the possibilities of correcting the changes associated with the persistence of long granulomatous inflammation.
The aim of the research. Explore the effect of various variants of anti-inflammatory therapy on the course
initiated by the SiO(2) granulomatous inflammation in the lungs of rats following the dynamics of macro - and microscopic changes in the respiratory system of experimental animals.
Subject and methods of research. The research was carried out on the basis of vivarium of the Higher state educational institution of Ukraine “the Ukrainian medical dental Academy”. The experimental part of the study, with the use of animals, carried out according to the compliance with the international principles of the European Convention «On the protection of vertebrate animals used for experimental and other scientific purposes (Strasbourg, 1985) and the standards of biomedical ethics, respectively Laws of Ukraine № 3446-IV 21. 02. 2006 year, Kiev, «On protection of animals from cruel treatment» with the conduct of medico-biological investigations, and ethical code of a doctor and a scientist.
The duration of the experiment was 8 weeks. Rats at a time received transthoracally a suspension of particles SiO(2) at the rate of 10 mg per 100 g of body mass in 0. 3 ml of physiological solution. The study included 72 Mature rats Wistar, mass 256,8 (228,2; 284,6) grams and age 7,2 (6,7; 7. 5) months. Of the 48 rats we formed four research groups of 12 individuals in each. One group of rats, in which the simulated GI, is left without treatment. Other research groups of animals with experimental pathology distributed taking into account the treatment - dexamethasone, glucosamine sulfate, or a combination of imunofani and hyaluronidase. The control group consisted of 24 rats were divided into 4 groups. In the part of rats of control group were not used any drugs. Number of animals without experimental pathology, depending on the group, received one of the following options: dexamethasone, glucosamine sulfate, a combination of imunofani and hyaluronidase. The input mode and the addition of drugs: dexamethasone (KRKA, Slovenia) of 0. 08 mg/kg, in/m, in 2 days; glucosamine sulfate (Dona Rottapharm Ltd. ” and “Sig-mar S. r. l. ” for “Rottapharm S. r. l. ”, Ireland/Italy)in the 5,33 mg/kg, in/m, in 2 days; a combination of imunofani (Bionoks (Russia, Moscow)on 0,67 mcg/kg, in/m and hyaluronidase (Lydaza-Biopharma, INC (Ukraine, Kiev) on 0,85 U/kg in/m, in 2 days. All of the above mentioned drugs at the time of their use in the study had the state registration and a full package of licensing documents in accordance with the current legislation of Ukraine. The duration of treatment ranged from 4 to 8 weeks
Fig. 1. Internally-alveolar cell productive inflammation. Color. hematoxylin and eosin. Magnification x 400.
in separate groups. Rats conducted complex clinical, laboratory, morphological, macroscopic examination. Withdrawn from experimental animal the tissues were prepared to light mycroscopy according to the conventional method [5]. Preparations were colored with hematoxyline and eozine. The analysis of macro - and
!3
(n=6) (n=6) (n=6) (n=6) (n=6) (n=6) (n=6) (n=6) (n=6)
4-th w eek of 8-th w eek of Treatment Treatment Treatment Treatment Treatment Treatment
the the from 4-th to 8- during 8 from 4-th to 8- during 8 from 4-th to 8- during 8
experiment experiment th week of weeks of the th week of weeks of the th week of weeks of the
Control group Groups of rats w ith a model Groups of rats with a model Groups of rats with a model groups of rats with a model
of the rats GI w ithout treatment of GI, which received a of GI, which received of GI, which received
without combination of imunofani and glucosamine sulfate dexamethasone
I Hyperinflation □ Emphysematous bullae □ Emphysema □ Inflammatory changes in interstitium of lungs □ Inflammatory changes in the bronch
Fig. 2. Comparison of macro - and microscopic changes in the lungs of rats with experimental modeling of granulomatous inflammation (GI) of the lung tissue compared with the control group.
Notes: 1. * - p<0. 05 for comparisons with the control group of the rats without treatment; 2. |= - p<0. 05 for comparisons between groups of rats with a model GI without treatment on the 4-th and 8-th week of the experiment; 3. | - p<0. 05 for comparisons with a group of rats with a model of GI without treatment on the 4-th week of the experiment; 4. + - p<0. 05 for comparisons with a group of rats with a model GI without treatment on the 8-th week of the experiment; 5. 1| -p<0. 05 for comparisons between groups of rats with a model of GI, which received glucosamine sulfate, a combination of imunofani and hyaluronidase with the group, where dexamethasone used from 4-th to 8-th week of experiment; 6. § - p<0. 05 for comparisons between groups of rats with a model of GI, which received glucosamine sulfate, a combination of imunofani and hyaluronidase with the group, where dexamethasone used during 8 weeks of the experiment; 7. # - p<0. 05 for comparisons between groups of rats with a model of GI, which received glucosamine sulfate and a combination of imunofani and hyaluronidase from 4-th to 8-th week of the experiment; 8. ^ - p<0. 05 for comparisons between groups of rats with a model of GI, which received glucosamine sulfate and a combination of imunofani with hyaluronidase during 8 weeks of experiment; 9. -f - p<0. 05 for comparisons between groups of rats with a model of GI that received medicamentous correction from 4-th to 8-week, and during 8 weeks of the experiment.
microscopic changes in the lungs of rats were held on the 4-th and 8-th weeks.
The comparison of the indicators was conducted using non-parametric methods [2, 3, 5]. Differences were considered statistically reliable at p<0,05
Research results and their discussion. For the qualitative analysis of the impact of treatment in rats of experimental group, first of all, we conducted a comprehensive analysis of the status of bodies of breath in animals of the control group. Among the intact animals of the control group of a significant level of macro- (hyperinflation, emphysematous bullae) and microscopic (emphysema, inflammatory changes in interstitium of lungs, inflammatory changes in the bronchi) of the pathological manifestations were observed. Also between the rats of control group which received dexamethasone, glucosamine sulfate, a combination of imunofani and hyaluronidase, an increase in the level of abnormal changes in the lungs is not detected. These results give us an opportunity to speak about the safeness of used drugs in relation to the lungs and allows us to use these data as a “standard rules” in comparison with animals that have artificially caused GI.
Transthoracal use of the suspension particles SiO(2) in rats was accompanied by focal intraalveolar granulomatous reaction (fig. 1).
The above mentioned histological preparation of the lung tissue of a rat (fig. 1), colored with hematoxylin and eosin, demonstrates the center of chronic productive inflammation. Cellular infiltration of the latter is represented by numerous macrophages, epithelioid cells, which sometimes merge, lymphocytes and single eosinophils. In addition, there is a heightened activity of fibroblasts that produce collagen fibers. The wall of the alveoli, which surrounds the cell, with the perceptible hyperemia, infiltration of macrophages and lymphocytes. The last is partly destroyed, that demonstrates the phenomenon of emphysema (Fig. 1).
The following stage of the work was to analyse macro - and microscopic changes of the respiratory organs of rats with induced SiO(2) granulomatous inflammation in the lungs depending on the received anti-inflammatory therapy (fig. 2).
It is worth mentioning that among the animals of the experimental group, where the artificially initiated granulomatous inflammation in the lungs on the 4-th week of the experiment it was noted significantly (p<0. 05) higher levels of inflammatory changes as in the bronchial tubes,
* *
2
and in the interstitial tissue of the lungs compared with the control group. On the В-th week of the experiment the animals with experimental pathology, that did not receive treatment significantly (p<0. 05) increased the number of animals, which showed the effects of hyperinflation (increased airiness of the lungs), and microscopically - emphysema, inflammatory changes in interstition of the lungs (fig. 2). Among the animals with the initiated granulomatous inflammation, receiving glucosamine sulfate, dexamethasone and a combination of imunofani and hyaluronidase in general, there was a significantly lower level of both macro- and microscopic changes in the lungs compared with the control group. Among the drugs that have been administered with the 4-th week of the experiment the most effective was dexamethasone. However, only under condition of В weeks’ use of dexamethasone in rats we could observe significantly (p<0. 05) lower than the level of evidence of increased airiness of the lungs (hyperinflation), compared with rats on the В-th week of the experiment without treatment.
So, as a result of the summary analysis was established that among all the applied options for the correction of the treatment, the best protective properties were found in the dexamethasone, provided appointment within В weeks of the experiment.
Conclusions. The use of drugs with anti-inflammatory properties (dexamethasone, glucosamine sulfate, a combination of imunophan and hyaluronidase) in rats with experimental initiated by the SiO(2) granulomatous inflammation accompanied by a lower level of pathological macro - and microscopic changes in the organs of respiration, compared with animals without treatment.
None of the applied options for correcting therapy had no advantages in comparison with each other for the anti-inflammatory properties.
The lowest level of the pathological changes in the lung tissue of rats initiated with experimental pathology was observed in the В weeks of the use of dexamethasone.
Prospects of the further researches. Further work in this direction will allow to improve intervention strategy in correction of granulomatous inflammation.
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УДК бІб. 24-002-092. 9-0В
АНАЛІЗ ВПЛИВУ РІЗНИХ ВАРІАНТІВ ПРОТИЗАПАЛЬНОЇ ТЕРАПІЇ НА ПЕРЕБІГ ІНІЦІЙОВАНОГО SiO(2) ГРАНУЛЕМАТОЗНОГО ЗАПАЛЕННЯ У ЛЕГЕНЯХ ЩУРІВ Бойко Д. М., Бойко М. Г., Ніколенко Д. Є.
Резюме. Дане наукове дослідження присвячене вивченню впливу різних варіантів протизапальної терапії на перебіг індукованого SiO (2) гранулематозного запалення у легенях щурів за динамікою макро- та мікроскопічних змін системи органів дихання експериментальних тварин.
За результатами дослідження було виявлено, що найменший рівень патологічних змін в тканині легень щурів, з ініційованою експериментальною патологією, спостерігався при В-тижневому використанні дексаметазону.
Ключові слова: гранулематозне запалення, SiO (2), щури, протизапальна терапія.
УДК бІб. 24-002-092. 9-0В
АНАЛИЗ ВЛИЯНИЯ РАЗЛИЧНЫХ ВАРИАНТОВ ПРОТИВОВОСПАЛИТЕЛЬНОЙ ТЕРАПИИ НА ТЕЧЕНИЕ ИНИЦИИРОВАННОГО SiO (2) ГРАНУЛЕМАТОЗНОГО ВОСПАЛЕНИЯ В ЛЕГКИХ КРЫС Бойко Д. Н., Бойко Н. Г., Николенко Д. Е.
Резюме. Данное научное исследование посвящено изучению влияния различных вариантов противовоспалительной терапии на течение иницированного SiO (2) гранулематозного воспаления в легких крыс по динамике макро- и микроскопических изменений органов дыхания экспериментальных животных.
По результатам исследования было выявлено, что наименьший уровень патологических изменений в тканях легких крыс с инициированной экспериментальной патологией наблюдался при В-недельном использовании дексаметазона.
Ключевые слова: гранулематозное воспаление, SiO (2), крысы, противовоспалительная терапия.
UDC б1б. 24-002-092. 9-0В
An Analysis of the Different Variants’ Impact of Anti-Inflammatory Therapy on the Course Initiated by the SiO(2) Granulometous Inflammation in the Lungs of Rats Boyko D. M., Boyko M. G., Nikolenko D. E.
Summary. This research study investigates the influence of different options of anti-inflammatory therapy on the course of initiated by the SiO (2) granulomatous inflammation in the lungs of rats by the dynamics of macro- and microscopic changes of the respiratory system in experimental animals.
The study found that the lowest level of pathological changes in the lung tissue of rats with experimental pathology observed at В weeks of using dexamethasone.
Key words: granulomatous inflammation, SiO (2), rats, anti-inflammatory therapy.
Стаття надійшла 25. 01. 2013 р.
Рецензент - проф. Гасюк А. Ї.
