Ultrastructure of alveolar macrophages in case of experimental acute renal failure

Zaiats L.M.; Klishch I.P.

СТРУКТУРЫ ЗМ1НИ ЕНДОКРИННО1 СИСТЕМИ СЕРЦЯ ПРИ СТРЕПТОЗОТОЦИНОВОМУ ЦУКРОВОМУ Д1АБЕТ1 Журакiвська О.Я., Микулець Т.И., Голдак У.М., Клипич Я.И., Мисьюв В.А, Гречин А.Б., Клипич О. О. Метою роботи було встановлення особливостей структурно! перебудови секреторних передсердних кардюмюциив у раннi та вщдалеш термiни перебiгу стрептозотоцинового цукрового дiабету (ЦД). ЦД моделювали одноразовим внутршньоочеревинним введенням стрептозотоцину (6 мг на 100г маси тша). Матерiал для дослщження забирали на 14 та 56 доби експерименту. Використали електронно-мкроскошчний метод дослiдження. Встановлено, що стрептозотоциновий ЦД в секреторних передсердних кардюмюцитах призводить до перебудови внутрiшньоклiтинних органел, яю вiдповiдають за синтез i секрецiю передсердного натрiйуретичного пептиду (ПНУП). Слiд зазначити, що вiдбуваeться перерозподш рiзних типiв секреторних гранул (СГ) у вщповщь на гiперглiкемiю, при цьому на 14 добу експерименту значно збшьшуеться об'емна щiльнiсть дифундуючих СГ, що вказуе на посилення процесiв виведення ПНУП iз ттини, а на 56 добу об'емна щшьшстъ молодих i зрiлих СГ достхдарно зменшуеться, що свiдчить про зрив компенсаторних механiзмiв.

Ключов1 слова: стрептозотоциновий цукровий дiабет, щурi, секреторний передсердний кардiомiоцит.

Стаття надiйшла 6.11.2017 р.

DOI 10.26.724 / 2079-8334-2018-1-63-130-133 ЦОС 616-092.9+616.24+616.61-008.6+616-08

СТРУКТУРНЫЕ ИЗМЕНЕНИЯ ЭНДОКРИННОЙ СИСТЕМЫ МИОКАРДА ПРИ СТРЕПТОЗОТОЦИНОВОМ САХАРНОМ ДИАБЕТЕ Жураковская О.Я., Микулец Т.И., Голдак У.М., Клипич Я.И., Мискив В.А., Гречин А.Б., Клыпыч О. О.

Целью работы было определение особенностей структурной перестройки сердечных предсердных кардиомиоцитов в ранние и отдаленные сроки течения експериментального стрептозотоцинового сахарного диабета (СД). СД моделировали одновременным внутрибрюшинным введением стрептозотоцина (6 мг на 100г массы тела). Материал для исследования изымали на 14 и 56 день експеримента. Использовали електронно-микрос копический метод исследования. Определено, что стрептозотоциновый СД в секреторных предсердных кардиомиоцитах ведет к перестройке внутриклеточных органэлл, что отвечают за синтез и секрецию предсердного натрийуретического пептида (ПНУП). Следует отметить, что происходит перераспределение разных типов секреторных гранул (СГ) в ответ на гипергликемию, при этом, на 14 сутки експеримента значительно увеличивается обьемная плотность дифундирующих СГ, что указывает на усиление процессов выведения ПНУП из клеток, а на 56 сутки обьемная плотность молодых и зрелых СГ достоверно уменьшается, что свидетельствует о срыве компенсаторных механизмов.

Ключевые слова: стрептозотоциновый сахарный диабет, крысы, секреторный предсердный кардиомиоцит.

Рецензент Срошенко Г.А.

ULTRASTRUCTURE OF ALVEOLAR MACROPHAGES IN CASE OF EXPERIMENTAL ACUTE

RENAL FAILURE

e-mail: ira181281@ukr.net

We have done experiments on Vistar line white male rats using electronic microscope method and studied in dynamics (12, 24, 72 hours) the ultrastructural changes of alveolar macrophages in case of experimental acute renal failure. It has been established that already in 12 hours after beginning of the experiment one can observe increase of quantity and functional activity of macrophage cells. With expansion of experimental timeline (24-72 hours), we have observed both dystrophic-destructive and adaptive changes in the alveolar macrophages.

Key words: lung, alveolar macrophages, experimental acute renal failure

The paper is a fragment of RSW "Pathogenetic Development Mechanisms of Changes in the Respiratory, Endocrine, Nervous Systems in Case of Simulated Pathological Conditions and Correction of Thereof" (number of state registration 0117U001758).

It has been established in the multiple clinical and experimental studies that alveolar macrophages (AM) play an important role in support of resistance of human body in case of exposure to exo- and endogenic factors [1, 2, 4, 6]. Analysis of many works has demonstrated that morphofunctional condition of AMs is closely connected with structural and metabolic changes in lungs in case of different pathological conditions [3, 5, 7, 10].

The purpose of this research was to study in dynamics the ultrastructural changes of alveolar macrophages of the respiratory part of lungs in case of experimental acute renal failure (EARF).

Materials and methods. The experiment was done on 45 Vistar line white male rats weighting 180220 grams, which were subdivided in two groups: control and experimental. Acute renal failure in rats of the experimental group was induced by intramuscular administration of 50% glycerol aqueous solution in quantity of 10 ml per kg of body mass [14]. Equivalent amount of water for injections has been injected to the control group. Lung tissue sampling for electronic microscope examination was done using ketamine anaesthesia in 12, 24, 72 hours after beginning of the experiment. Pieces of lung tissue were fixed in 2,5% solution of gluteraldehyde with further postfixation in 1% solution of osmium tetroxide. After dehydration, the material

was poured over epon araldite. The cuts obtained on ultramicrotome "Tesla BS-490" were studied using electron microscope "PEM-125K".

Results and their discussion. Electronic microscope examination of the respiratory part of lungs in 12 hours after beginning of the experiment has demonstrated increase of AMs quantity in the air cells to 3,88±0,048 (p<0,001, table 1) as compared with control group. The nuclei of macrophage cells have incorrect form with evenly distributed chromatin. The nuclear membrane has distinct sinuous contours and forms superficial invaginations. In cytoplasm, we observe mitochondria of different size and form with matrix of moderate electronic-optical density. At the same time, we note a significant quantity of small lysosomes and phagosomes different in form, size and content. Golgi apparatus (GA) consists of small blisters and vacuoles.

Table 1

Quantity of alveolar macrophages in lung tissue of white rats in case of experimental acute renal failure

Groups of animals Statistical index Periods of monitoring

12 hours 24 hours 72 hours

Control M ± m 2,04±0,06 2,16±0,06 2,24±0,03

Experimental M ± m 3,88±0,05 5,24±0,08 4,30±0,05

P <0,001 <0,001 <0,001

The tubules and cisterns of granular endoplasm grid (GEG) are somewhat dilated with tender fibrous osmiophil content. On the external membrane of the latter are found ribosomes. The plasma membrane of AMs creates a big quantity of cytoplasm bulges. In some cells are defined mitochondria increased in volume with reduced cristae. As per experiment continuance (24 hours), the AMs quantity has significantly increased as compared with control and was 5,24±0,083 (p<0,001, see Table 1). At the present stage of the experiment, we could determine jjolymorphism of macrophage elements in the alveolar lumen (Fig. 1).

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Fig 1. Ultrastructural heterogeneity of alveolar macrophages in 24 hours after beginning of the experiment. 1 - alveolar lumen; 2 - alveolar macrophages; 3 - fragment of type II alveolocytes. Electronic mictophotography x4000.

Fig. 2. Ultrastructural organization of the alveolar macrophages in 72 hours after beginning of the experiment. 1 - alveolar lumen; 2 - nucleous; 3 - mitochondria; 4 - lysisome; 5 - phagosome; 6 - hemocapillary lumen. Electronic microphotography. x6400.

Submicroscopically, AMs differ in size, form and ultrastructural organization. Many AMs have nuclei with small-grained nucleoplasm of low electronic optical density. The nuclear membrane forms superficial invaginations. The paranuclear space is locally dilated. Mitochondria have matrix of moderate electronic-optical density and some fragmented cristae. The elements of GA and GEG are dilated. The quantity of ribosomes on the external membrane of the latter is reduced. In some cells, we observe fragmentation of GEG membranes. In cytoplasm, one can note insignificant quantity of lysosomes and some big phagosomes containing polymorphous osmiophil material of different dimensions. On the apical surface of such cells can be identified cytoplasm growing cell membranes small in size. Among AM population, at the level of dystrophic - destructive changes one can observe some cells with characteristic features of increased functional activity (Fig 1.). The nuclei of such cells have matrix of average electronic-optical density. The chromatin granules are evenly placed on the whole area of the nuclei. In the cellular cytoplasm is observed well expressed synthetical apparatus represented by the cisterns of Golgi apparatus and hypertrophic GEG tubules with multiple ribosomes on the membranes of thereof. Mitochondria have matrix of moderate electronic-optical density and are of different size and form. In cytoplasm of AMs can be noted a significant quantity of lysosomes and phagosomes different in form, dimensions and structure. In 72 hours after beginning of the experiment, the quantity of macrophage elements continues increased as compared with control, and is 4,30±0,050 (p<0,001). However, in comparison with the previous stage of the experiment, AMs quantity is reduced (see Table 1.).As on the previous stage of the experiment, heterogeneity of AMs is observed in the alveolar lumens. Among some scanty actively phagocytising AMs are observed the cells with dystrophic and destructive changes (Fig. 2). The nuclei of macrophage elements are deformed. Nucleoplasm

has matrix of low electronic optical density. The granules of chromatin are located along the external surface of nuclear membrane or sometimes appear grouped into separate clusters. Mitochondria are swollen, with shortened and disoriented cristae. The cisterns and tubules of GA and GEG are dilated and vacuolated. In cytoplasm, we observe isolated lysosomes and phagosomes with visible lamellar bodies and fragments of destroyed cells. The study carried out has shown that already after 12 hours of simulation of acute renal failure one can observe increase of quantity and functional activity of AMs in the alveolar lumens. Our data agree with the research results of other scientists that point to the fact that AMs are one of the most reactive elements of lung tissue in case of exposure to different exogenic and endogenic factors [1, 4, 11]. It is evident that such increase of quantity and functional activity of macrophage elements can be considered as primary response of AMs to lung tissue lesion [4, 10, 13]. As per experiment continuance (24 - 72 hours), in the alveolar lumens along with active phagocytising macrophage cells are observed AMs with some lysosomes and increased quantity of big phagosomes that prove the functional deficiency of macrophages. Changes of similar nature in submicroscopic organization of AMs under the influence of exogenic and endogenic factors are reported by other scientists as well [2, 8, 9, 12].

1. Our research has demonstrated that experimental acute renal failure is accompanied by the expressed changes of submicroscopic structure of alveolar macrophages.

2. The nature and degree of expression of ultrastructural changes in alveolar macrophages depends on duration of endogenic factor exposure.

Prospects for further research. The study ofphagocyte activity of alveolar macrophages in case of experimental acute renal failure is in the perspective of future research.

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