Morphological and morphometric rearrangements of the rat adenohypophysial-thyroid system under the experimental extracellular dehydration Текст научной статьи по специальности «Фундаментальная медицина»

тсля введення ХС на т токсично! да ДГ. Вiдновлювалася структура дшянок плазматично! мембрани джгутиюв, матрикс мтохондрш був електронно-щiльний, спостерiгалося утворення крист i дше'шових нiжок близько дупле™ микротрубочек, що свiдчить про забезпечення нормально! рухливост сперматозо!дiв.Пiд впливом ХС здшснювалося поступове вiдновлення репаративних процеав в клiтинах сперматогенезу щурiв, яю зазнали впливу ДГ.Ключовi слова: ультраструктура ам'яниюв, хондро!тину сульфат, доксорубiцину пдрохлорид.

Ключовi слова: ультраструктура сiм'яникiв, хондро!тину сульфат, доксорубiцину гiдрохлорид.

Стаття надшшла 08.04.19 р.

полноценности сперматозоидов, то есть начала восстановительных процессов после введения ХС на фоне токсического действия ДГ. Восстанавливалась структура участков плазматической мембраны жгутиков, матрикс митохондрий был электронно-плотным, наблюдалось образование крист и динеинових ножек около дуплетов микротрубочек, что свидетельствует об обеспечении нормальной подвижности сперматозоидив.Под влиянием ХС осуществлялось постепенное восстановление репаративных процессов в клетках сперматогенеза крыс, подвергшихся воздействию ДГ.

Ключевые слова: ультраструктура семенников, хондроитина сульфат, доксорубицина гидрохлорид.

Рецензент Старченко 1.1.

DOI 10.26724/2079-8334-2019-2-68-174-179 UDC 616.441:616.151.1-092.9

N.B. Hry ntsov a, I.V . Klionieiiko, A.M. Romany uk, V .1. Buiiu-isier, I. A. Krav tsova

Medical Institute °f Sumy State Univ^sity, Sumy

MORPHOLOGICAL AND MORPHOMETRIC REARRANGEMENTS OF THE RAT ADENOHYPOPHYSIAL-THYROID SYSTEM UNDER THE EXPERIMENTAL

EXTRACELLULAR DEHYDRATION

e-mail: natalia.gryntsova@gmail.com

Disruption of homeostasis cause significant changes in all the body systems, including the endocrine one. The purpose of the work is to study the morphological and morphometric rearrangements of the adenohypophyseal-thyroid axis in rats under conditions of experimental moderate degree extracellular dehydration. The experiment was performed on 12 rats of control and experimental group. The experimental group's rats for 60 days modeled the moderate degree of extracellular dehydration. Histological, morphometric and statistical methods of investigation were used. Disturbance of the water-salt balance of the body causes a slight increase in secretory activity on the part of adenohypophysis and the development of hypofunction of the thyroid gland, which negatively affects the development of compensatory and adaptive processes in the body.

Key words: adenohypophysis, extracellular dehydration, thyroid gland, stress

The work is a fragment of the eresearch projects "Morphofunctional aspects of the body homeostasis" (state registration No. 0118U006611)

Homeostasis provides the necessary conditions for the life of both individual cells and the body as a whole. Constancy of water-electrolyte composition is part of homeostasis [7, 12]. In forming the regulation of the compensation mechanism for various extreme factors affecting the body, the endocrine system plays one of the key roles [4, 14]. Adequacy, nature of the body's adaptive changes depends on changes in the hormonal secretion that provide the body homeostasis [5]. Steady and multiple stress can be the basis of stress-induced pathology development [5, 15]. The pituitary - thyroid axis of the endocrine system, together with the pituitary gonadal, sympathoadrenal and hypothalamic -pituitary adrenocorticular systems, play a key role in providing the adaptive response of the body to stress effects. The thyroid gland's hormones are known to take an active part in the development of adaptive responses and have the ability to increase the sensitivity of adrenoreceptors to adrenaline and noradrenaline [5]. Conditions associated with changes in the activity of regulatory stress systems (in acute or chronic stress, post-traumatic stress disorders, changes in physical activity), also affect the functioning of the hypothalamic-pituitary-thyroid axis of the endocrine regulation [5]. To date, the thyroid gland's status has been studied under the influence of heavy metals salts [6], moderate chronic hyperthermia [11], geochemical factors [10], tiotriazolini [5], and antipsychotic therapy depending upon the age [1]. According to the results of the literature sources search, there is no works on studying morphological and morphometric indices of adenohypophysis blood vessels in rats under the conditions of experimental moderate degree extracellular dehydration.

The purpose of the study was to study morphological and morphometric rearrangements of the adenohypophysial-thyroid axis structural components in sexually mature male rats under the conditions of experimental extracellular dehydration.

© N.B. Hryntsova, I.V. Khomenko, 2019 174

Materials and methods. The experiment was carried out on 12 white, sexually mature male rats weighing 230-300 g, divided into 2 groups (control and experimental). Animals of the experimental group were modeled the moderate degree of extracellular dehydration for 60 days [9]. Groups of experimental animals were sacrificed by decapitation under etheric anesthesia in compliance with the provisions of the General Ethical Principles of Animal Experiments (Kyiv, 2001). The thyroid gland was identified and prepared according to [8]. The established techniques of histological research method were applied. The sections were stained with hematoxylin-eosin and applying the Schick reaction (PAS-periodic acid-Schiff stain). The general morphological and morphometric analysis was performed using "Zeiss Primo Star" light optic microscope, with lenses x10, x20, x40 and binoculars 7, 10. For the morphometric study of micropreparations, the "SCPR-2017-Zen 2 lite" software was used with photodocumentation of the results by means of "axiocam ERC 5S Zeiss" digital camcorder. The mean values of the morphometric parameters for the thyroid gland were determined: the larger and smaller follicle diameters, the follicles and colloid area, the thyroid epithelium area, the number of tyrocytes in the follicle, the height of the tyrocytes, the larger and smaller diameters and the area of the follicular epithelium nuclei, the colloid accumulation index (CAI) and follicular colloidal index (FCI) [3, 6].

The following morphometric indices were used to determine the morphofunctional features of the adenohypophysis endocrinocytes: large and small diameters of adenocytes bodies and nuclei, the area of adenocytes transsection and their nuclei, the area of the endocrinocyte cytoplasm, the nuclear-cytoplasmic ratio, and the mean karyon diameter (MKD) of adenocytes [2]. Morphometric indices characterizing the vascular bed of adenohypophysis were studied in earlier papers by the authors [3]. Statistical processing of the obtained results was carried out with a personal computer using the "Statistica 8.0" software.

Results of the study and their discussion. On the 60th day of the experiment, it was found that disorders in the vascular component of the thyroid gland and adenohypophysis are similar, prevalent and primary, more pronounced in the parenchyma of adenohypophysis. Vascular disturbances in the form of diffuse venous-capillary hypertrophy of vessels with significant and moderate degree of blood filling and changes in rheological properties of blood (stasis, with initial stages of sludge and thrombogenesis) were determined. The vascular wall permeability increases with formation of individual small-focal diapedetic microhemorrhages (fig. 1, 3). Microcirculation disorders in the adenohypophysis and the thyroid gland are of nonspecific, polymorphic nature. Greater severity of vascular disorders in the adenohypophysis, in our opinion, is explained by the features of the anatomical blood supply of this internal secretion gland [3] (fig. 1). In our opinion, pathogenetical dystrophic rearrangements of the endocrine glands' vascular bed including the vascular wall (hypoxic dystrophy of the endothelial cells) have a negative effect on the morphofunctional and secretory activity of the organs' cells.

Thus, a significant congestion of the adenohypophysis causes a disorder of the gland's normal trabecular structure. Disorganization and discoupling of epithelial trabeculae are observed. Around the enlarged hemocapillars with erythrostasis, groups of adenocytes with slightly hyperchromic cytoplasm and hyperchromic nuclei are found. In other microscope fields of preparations, hypo- and normochromic cells prevail (fig. 1).\

Fig.1. Morphological rearrangements of the vascular bed and Fig.2. Thyroid gland of a sexually mature rat under the

cells of the adenohypophysis under the conditions of the moderate conditions of the moderate degree experimental extracellular

degree experimental extracellular dehydration are observed. dehydration: Schick-positive colloid; stromal swelling. Staining:

Staining with hematoxylin-eosin. x 400x. Schick test x 400.

The secretory cells nuclei can both maintain an oval, rounded form and acquire an elongated, oblong form. Chromatin nucleoreticulum with fine-dispersed condensation of chromatin is diffusely located throughout the nucleus. However, a small part of the nuclei still has a hyperchromic hypertrophied nucleolus, which can be located both centrally and in a state of ectopia. The morphometric indices of adenocyte bodies are reduced somewhat unreliably in comparison with the control animals. Thus, the adenocyte bodies' large diameter index is reduced by 18.4% (p>0.05). The linear index of the cell bodies' small diameter is increased by 5.3% (p>0.05). The area of the adenocyte bodies transsection is reduced by 4.1% compared to the control animals. The area of the cells bodies cytoplasm transsection is reliably reduced compared to the control animals group by 14.2% (p<0.05, t = 3.150715). The mean morphometric indices of the adenocyte nuclei large and small diameters in experimental animals slightly exceed the control animals' indices by 11.6% and 6.7% respectively (p>0.05). The adenocyte nuclei transsection area exceeds that of the control animals by 11.5% (p>0.05). The mean diameter of the adenocyte karyon (MKD) of experimental animals exceeds the control animals' values by 10% (p<0.05, t = 2.828427). The nuclear-cytoplasmic ratio is reduced to 1:1.2 (table 1).

Table 1

Results of the morphometric study of adenogypophysis adenocytes in sexually mature rats under the moderate degree experimental extracellular dehydration conditions (X ± Sx)

Index Groups of laboratory animals

Control group rats Experimental group rats

Adenocyte body's large diameter, ^m 8.14±0.64 6.64±0.44

Adenocyte body's small diameter, ^m 5.49±0.24 5.78±0.42

Adenocyte bodies transsection area, ^m2 39.5±3.13 37.89±3.13

Adenocyte nuclei large diameter, ^m 4.3±0.22 4.8±0.2

Adenocyte nuclei small diameter, ^m 3.56±0.21 3.8±0.18

Adenocyte nuclei transsection area, ^m2 15.51±1.23 17.29±1.1

Adenocyte cytoplasm area, ^m2 24.0±0.58 20.6±0.91*

Nuclei-cytoplasm ratio 1:1.55 1:1.2

Mean karyon diameter (MKD) 3.9 4.3*

Note: difference between the control and experimental indices is: * < 0.05; ** p < 0.01; *** p<0.001.

On the 60th day of the study, in the thyroid gland's parenchyma, the size of the follicles increases, and the polymorphism of their shape and size from the irregular polygonal to the round shape is observed. Follicles are overexpanded by colloid, which becomes more condensed, granular, dense, better absorbs histologic stainers (fig.2). An increase in the amount of rough fibrous stroma is observed. Particles of the thyroid gland's parenchyma are clearly separated from each other by edematous and sclerotized stroma (fig.3).

In this case, the amount of extra follicular epithelium is significantly reduced, a large number of follicles with follicular wall rupture and reduced follicles are observed. In the site of the destroyed follicles, adaptive processes of replacing the parenchyma with the connective tissue are observed. In isolated fields of view, insignificant folliculogenesis from the remnants of the intrafollicular epithelium is observed. The predominant amount of tyrocytes acquires flat shape, the decrease in their height is observed. The thyrocytes' nuclei change are deformed, flattened, condensing, hyperchromic and homogeneous. A small portion of tyrocytes undergo destruction and desquamation into the lumen of the follicle. The process of tyrocytes proliferation is absent.

An increase in the follicles' larger and smaller diameters compared to the control group of animals was observed by 25.5% (p< 0.001, t = 9.54) and 17.1% (p< 0.001, t = 7.46), respectively. The follicles area is increased by 53.69% (p< 0.001, t = 16.46) compared to that of the control. The

Fig. 3. Thyroid gland of a sexually mature rat under the conditions of the moderate degree experimental extracellular dehydration: edema of the interparticle stroma; congestion and permeability disorder of the vascular wall with the erythrocytes release into the extravascular space; colloid condensing. Staining with hematoxylin-eosin. x 400x.

morphometric analysis of the follicles shows a significant, reliable increase in the colloid amount and in the follicular epithelium area. Thus, the colloid area is increased by 2.1 times (p< 0.001, t = 23.3), and the follicular epithelium area - by 9.1% (p<0.05, t = 2.38). The index of colloid accumulation in the experimental animals' follicle is by 55.2% (p< 0.001, t = 25.04) greater than that of the control animals. The follicular-colloid index, on the contrary, is reduced by 28.4% (p< 0.001, t = 29.63) compared to the respective index in the control animals. At the same time, the quantitative indices of the follicular epithelium remain virtually unchanged compared to the indices of the control group animals and exceed the latter by only by 1.6% (p>0.05). The height of tyrocytes in experimental animals is significantly reduced by 19.1% (p< 0.001, t = 6.71). The shape of the cells varies from cubic in the control animals to the flat in the experimental ones. Along with the decrease in the cell height, the reduction in the values of the tyrocytes nuclei larger diameter and the nuclei area by 21.7% (p< 0.001, t = 15.43) and 19.14% (p< 0.001, t = 4.93) respectively is observed. The smaller diameter of the nuclei increases slightly by 3.1% (p>0.05), which undoubtedly indicates that the experimental animals' nuclei acquire a more flat, elongated shape compared to the oval form of nuclei in the control animals (table 2).

Table 2

Results of the morphometric study of the thyroid follicles in sexually mature rats under the moderate degree experimental extracellular dehydration conditions (X ± Sx)

Index Groups of laboratory animals

Control group rats Experimental group rats

Larger follicles diameter, |im 52.43±0.96 65.79±1.02***

Smaller follicles diameter, ^m 43.76±0.67 51.26±0.75***

Follicles area ^m2 2194.34±49.95 3372.44±51.28***

Colloid area ^m2 989.73±32.25 2058.16±32.61***

Thyroid epithelium area, ^m2 1204.61±32.05 1314.28±33.12*

Thyrocytes height, |im 22.0±0.13 6.29±1.18***

Number of thyrocytes in the follicle 19.0±0.64 19.3±0.67

Larger nuclear diameter, ^m 4.15±0.03 3.25±0.05***

Smaller nuclear diameter, ^m 3.6±0.04 3.71±0.05

Thyrocytes nuclei area, ^m2 14.94±0.41 12.08±0.41***

CAI (colloid accumulation index) 3.37±0.54 5.23±0.051***

FCI (follicular colloidal index) 2.22±0.014 1.59±0.016***

Note: difference between the control and experimental indices is: * p<0.05; ** p<0.01; *** p<0.001.

Analysis of literature data showed that the hormones of the hypothalamic-pituitary-gonadal and thyroid axis of the endocrine regulation along with the sympathoadrenal and hypothalamic-pituitary-adrenal link of the endocrine regulation system play the key role in the response to extreme impacts of the environment and the body's adaptation. In the mechanisms of the body's homeostasis regulation, to provide adaptation in response to the stress effects, changes in hormonal secretion are a kind of cascade of closely interconnected reactions [13]. Under the influence of extracellular dehydration, on the background of general hypovolemic disorders in the adenohypophysis and the thyroid gland, microcirculation disorders of nonspecific polymorphic nature are developing. These disorders cause vessels blood filling disorders (diffuse venous-capillary congestion), changes in the blood rheological properties with initial stages of thrombogenesis, increase in vascular walls permeability with formation of fine-focal diapedetic microhemorrhages. Due to disorders in the blood rheological properties and blood supply (hemoconcentration, reduced mass of the circulating blood and the increased blood viscosity), the tissue hypoxia develops in the parenchyma of the gland. Pathogenetical dystrophic rearrangements in the endocrine glands vascular bed have a negative effect on the morphofunctional and secretory activity of glandular cells. However, a number of adenocytes nuclei morphological characters (the presence of hyperchromic hypertrophied nucleolus) and morphometric indices, including the adenohypophysis adenocytes' mean karyon diameter (MKD) in experimental animals, are slightly higher than those of the control animals. This undoubtedly indicates a certain increase in the functional activity and the strain of adaptive-adjusting processes in the adenohypophysis in response to the damaging agent's action. At the same time, in the parenchyma of the thyroid gland, pronounced signs of morphofunctional rearrangement are observed in the form of decreasing secretory and proliferative activity, focal reduction of follicles, uneven development of sclerotic changes. Morphologically, the decline in the gland's functional activity (hypofunctional state) occurs both due to disruption of the colloid utilization and due to reducing the synthetic activity of tyrocytes [3, 6]. This undoubtedly indicates a breakdown of physiological compensatory and adaptive processes with prolonged influence of extracellular

dehydration on the body of experimental animals. A vivid proof of the gland s reduced functional activity is reduction in the morphometric indices of the tyrocyte nuclei, the increased colloid accumulation index (CAI) and, at the same time, the reduced follicular-colloidal index (FCI).

The body's water-salt imbalance (the modeled moderate degree extracellular dehydration) causes negative changes in the structural components of the adenohypophysial-thyroid system of the experimental animals. Insignificant increase of the secretory activity and the stress of adaptive-adjustment processes on the part of adenohypophysis is not sufficient and effective to maintain homeostasis in the peripheral link of the endocrine system, i.e. thyroid gland. The development of the thyroid gland hypothyroidism undoubtedly affects the development of the compensatory and adaptive processes in the body as a whole and the course of the general adaptive syndrome in response to the damaging agent's action. This conclusion correlates with a number of authors' opinion about the participation of triiodo- and tetraiodothyronine in the most important body's metabolic reactions, including stress. In addition, one of the most important effects of thyroid hormones is their ability to provide a permissive effect on the body tissues' sensitivity to the action of catecholamines, increasing the sensitivity of adrenoreceptors to noradrenaline and adrenaline and increasing the density of some blockers on the cells' surface. Thus, according to the five-phase model of chronic stress, the adenohypophysial-thyroid axis of the endocrine system is at the stage of chronic stress "anxiety", which correlates with the opinion of other researchers.

Prospects for further developments are based on performing further morphological and immunohistochemical studies of the rat endocrine glands under the conditions of the body's water-salt balance disorders.

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Реферат

МОРФОЛОГ1ЧН1 ТА МОРФОМЕТРИЧН1

ПЕРЕБУДОВИ АДЕНОГ1ПОФ1ЗАРНО-ТИРЕО1ДНО1 СИСТЕМИ ЩУР1В ЗА УМОВ ЕКСПЕРИМЕНТАЛЬНО1 ПОЗАКЛ1ТИННО1 ДЕГВДРАТАЦП Гринцова Н.Б., Хоменко 1.В., Романюк А.М.,

Бумейстер В. I., Кравцова I.A. Порушення гомеостазу спричиняють значш змши в ycix системах оргашзму, в тому чи^ i ендокриннш. Метою роботи е вивчення морфолопчних та морфометричних перебудов аденоriпофiзарно-тиреоiдноi

МОРФОЛОГИЧЕСКИЕ И МОРФОМЕТРИЧЕСКИЕ

ПЕРЕСТРОЙКИ АДЕНОГИПОФИЗАРНО-ТИРЕОИДНОЙ СИСТЕМЫ КРЫС В УСЛОВИЯХ ЭКСПЕРИМЕНТАЛЬНОЙ ВНЕКЛЕТОЧНОЙ ДЕГИДРАТАЦИИ Гринцова Н.Б., Хоменко И.В., Романюк А.М., Бумейстер В. И., Кравцова И. А. Нарушение гомеостаза вызывает значительные изменения во всех системах организма, в том числе и эндокринной. Целью работы является изучение морфологических и морфометрических перестроек

вiсi статевозрших щурiв в умовах експериментально! позаклтнно! депдратацп середнього ступеня. Експеримент проведений на 12 щурах контрольно! та експериментально! групи. Щурам експериментально! групи протягом 60 дiб моделювався середнш стутнь позаклiтинного зневоднення. Застосовувалися пстолопчш, морфометричнi та статистичнi методи дослщження. Порушення водно-сольового балансу оргашзму викликае незначне пiдвищення секреторно! активност зi сторони аденогiпофiзу та розвиток ппофункци щитоподiбно!' залози, що негативно впливае на розвиток в органiзмi компенсаторно-

пристосувальних процесiв.

Ключовi слова: аденогiпофiз, позаклiтинна дегiдратацiя, щитоподiбна залоза, стрес.

Стаття надiйшла 29.06.18 р.

аденогипофизарно-тиреоидной оси половозрелых крыс в условиях экспериментальной внеклеточной дегидратации средней степени. Эксперимент проведен на 12 крысах контрольной и экспериментальной группы. Крысам экспериментальной группы в течение 60 суток моделировалась средняя степень внеклеточного обезвоживания. Применялись гистологические,

морфометрические и статистические методы исследования. Нарушение водно-солевого баланса организма вызывает незначительное повышение секреторной активности со стороны аденогипофиза и развитие гипофункции щитовидной железы, что отрицательно влияет на развитие в организме компенсаторно-приспособительных процессов.

Ключевые слова: аденогипофиз, внеклеточная дегидратация, щитовидная железа, стресс.

Рецензент Срошенко Г. А.

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В роботi проведене дослiдження судин мкроциркуляторного русла обмiнноI ланки часточок тднижньощелепно! залози щурiв. За результатами дослщжень встановлено що капiляри часточок залози реагують на дiю 1 % розчину метилового ефiру метакрилово! кислоти стiйкою дилятацieю протягом експерименту та тдтверджуеться збiльшенням дiаметру зовншнього та дiаметру просвiту на 29,50 % (р> 0,05), на 14 добу, та на 17,99 % (р> 0,05) на 30 добу, iз зменшенням iндекса Вогенворта на 31,44 % порiвняно з контрольною групою. Вiдновлення показникiв до юнця експерименту не вiдбуваeться внаслiдок постшно! токсично! ди ефiру метакрилово! кислоти.

Ключов! слова: капшяр, шднижньощелепна слинна залоза, метакрилова кислота, щури.

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Робота е фрагментом НДР «Експериментально-морфологiчне вивчення ди трансплантатiв крюконсервованог плаценти та тших екзогенних чиннитв на морфофункщональний стан ряду внутрштх оргатв», номер державно! реестрацн № 0113и006185.

Метилметакрилат, як матер1ал, широко використовують для базишв протез1в, при виготовленш ортодонтичних апарапв. [2]. У науковш л1тератур1 наявш багаточисельш пращ, яю присвячеш перевагам { недолшам акрилових пластмас, як використовуються для виготовлення базис1в зшмних протез1в [6].

Морфолопчне дослщження змши слизово! оболонки порожнини рота статевозрших бших щурах лшй Вистар шд впливом метилметакрилату показало, що при тривалому вплив1 метилметакрилату на слизову оболонку порожнини рота, спостерйаються структурш змши в ус1х шарах слизово!, що формують розвиток токсичного запального процесу. Вщбуваються дегенеративш змши ештелюципв, ознаками репарацй ештел1ю, що виявляеться його пперплазда, акантозом { фокальним гиперкератозом. У тдслизовому шар1 в частит випадюв виявляються ознаки негранулематозного запалення. Отже, вважають фах1вщ, при зубному протезуванш з використанням акрилових пластмас необхщно враховувати можливу токсичну д1ю метилметакрилату { проводити заходи щодо зменшення цього впливу [5].

При вивченш оргашв та систем необхщно враховувати значний вплив стану ланок гемомшроциркуляторного русла на !х функцюнування - особливо обмшно!, яка забезпечуе повноцшний газообмш м1ж кров'ю та кл1тинами, та забезпечення !х необхщними для життед!яльносп речовинами.

При дй 1 % розчину метилового еф1ру метакрилово! кислоти спостерйаеться спазм резистивно! ланки мшроциркуляторного русла шднебшних залоз щур1в на 14 добу спостереження, на замшу якому визначаеться дилятащя до 30 доби експерименту. З боку обмшно! 1 емшсно!

© Г.А. Срошенко, Д.Р. Крамаренко,, 2019