CC BY
16
запального вогнища у пародонтальному комплекса Зокрема, nepe6ir експериментального пародонтиту супроводжувався пiдвищенням вмюту алексин-позитивних (раннiй апоптоз) та некротизованих нейтрофшв, що пов'язано з посиленням штенсивност ix утворення у вщповщь на антигену стимуляцiю. При данш модельованiй патологи реалiзацiя шдуковано! смертi клiтин вiдбувалася переважно шляхом апоптозу.
Ключовi слова: Пародонтальний комплекс, запалення, нейтрофiльнi гранулоцити, апоптоз, некроз.
Стаття надшшла 16.01.18 р.
очага в пародонтальном комплексе. В частности, ход экспериментального пародонтита сопровождался повышением содержания анексин-положительных (ранний апоптоз) и некротизированных нейтрофилов, что связано с усилением интенсивности их образования в ответ на антигенную стимуляцию. При данной моделируемой патологии реализация индуцированной смерти клеток происходила преимущественно путем апоптоза.
Ключевые слова: Пародонтальный комплекс, воспаление, нейтрофильные гранулоциты, апоптоз, некроз.
Рецензент Денефшь О.В.
DOI 10.26724 / 2079-8334-2018-2-64-142-145 UDC 616-092+616.24+616.37-002+616-08
ULTRASTRUCTURAL CHANGES IN PULMONARY HEMOMICROCIRCULATION AT ACUTE EXPERIMENTAL PANCREATITIS
E-mail: [email protected]
Experiment performed on 70 white male rats of the Vistar line by electron microscopic method, dynamics (12, 24, 48, 72 h) of ultrastructural changes of the hemomycocirculatory bed of lungs in experimental acute pancreatitis was studied. Established, that at 12 h after the beginning of the experiment there are violations of the submicroscopic structure of the hemomycocirculatory bed of the lungs. In hemocapillaries of the alveolar wall, excessive accumulation of leukocytes, their adhesion and aggregation is noted. With the duration of the study (24-72 h) in the hemomicrocirculatory bed of the lungs are defined as dystrophic-destructive as compensatory-adaptive changes.
Keywords: lungs, hemomicocirculatory bed, acute experimental pancreatitis.
The present study is a fragment of the research work "Pathogenetic Development Mechanisms of Changes in the Respiratory, Endocrine, Nervous Systems in Case of Simulated Pathological Conditions and their Correction " (number of state registration 0117U001758).
Numerous clinical and experimental studies have shown that lungs are one of the first target organs that can be damaged in various critical states (sepsis, polytrauma, acute pancreatitis, peritonitis, acute renal failure, thermal injuries, acute blood loss) [8,7,9,11,14]. At the same time, under the influence of various extreme factors there is a violation of the morphofunctional state, constituent components of the aerohematous barrier, which underlies the development of syndrome acute lung injury (ALI) [1,4,5,13].
The purpose of the research was to study in dynamics hemomicrocirculatory bed of the lungs in acute experimental pancreatitis.
Materials and methods. Experiments were performed on 70 white male rats weighing 180-220 g. Animals were divided into 3 groups: I - Intact group of animals (n = 10); II - control (n = 20); III - with a model of acute pancreatitis (n = 40). All studies were performed under general anesthesia using ketamine (40 mg / kg). Animal retention and manipulation were carried out in accordance with the provisions of the Law of Ukraine "On the Protection of Animals from Cruelty" (No. 1759-VI of 15.12.2009). Acute experimental pancreatitis was reconstructed by two intraperitoneal injections of 20% solution of L-arginine in a total dose of 5 g/kg with a one-hour interval [13]. Control group of animals was injected equivalent dose of physiological solution. Pulmonary tissue collection for electron microscopic examination was performed under ketamine anesthesia after 12, 24, 48 and 72 hours. The material was fixed in 2.5% glutaraldehyde solution, followed by fixation in a 1% solution of osmium tetrachloride. After dehydration, the material was poured into epon-aralgit. Cuts obtained on ultramicrotome "PEM-125K".
Results of the study and their discussion. An analysis of the results of an electron microscopic study showed that after 12 h after the beginning of the experiment in the hemocapillaries of the alveolar wall, an increased number of neutrophilic leukocytes, their aggregation and adhesion to the endothelial cells (Fig. 1) is observed. Endothelial cell nuclei with a matrix of mean electron-optical density. The chromatin granules are substantially uniformly distributed throughout the core area. The nucleolem has winding contours and forms a shallow invagination. At the same time, separate endothelial cells with low electron-optical density nuclei and marginal distribution of chromatin granules are noted. Mitochondria of different size and shape with an enlightened matrix and partially reduced crests. Cisterns and channels
© L.M. Zaiats, V.V. Cherkasova, 2018
of the Golgi apparatus (GA) and rough endoplasmic reticulum (RER) are unevenly expanded. In the peripheral cytoplasmic sites, both small and large microvesicles are observed. On the luminal surface of some endothelial cells there are microvessels that protrude into the lumen of hemocapillaries.
Fig. 1. Adhesion of leukocytes to the endothelium of the hemocapillary wall of the alveoli after 12 h of experiment. Marking: 1 - lumen of hemocapillary; 2 - peripheral part of the endothelial cell; 3 - neutrophilic leukocyte; 4 - lumen of the alveoli; 5 -interstitium. Electronic microphotography. Magn.: x9600.
Fig. 2. Ultrastructure organization of the hemocapillary wall of the alveoli in 24 h after the experiment. Markings: 1 - lumen of hemocapillary; 2 - sail-like protrusion of the plasma of the endothelium in the lumen of the hemocapillary; 3 - platelet; 4 - lumen of the alveoli. Electronic microphotography. Magn.: x6400.
24 hours after the beginning of the experiment in the lumen of many hemocapillaries, red blood cells are detected, adhesion and aggregation of leukocytes and platelets. In the lumen of individual hemocapillaries, along with cellular elements, amorphous, nonstructural, electron-dense masses and destructively altered fragments of organelles are detected. Endothelial cells with matrix of low electron-optical density. Granular chromatin concentrates near the nuclear membrane. Nuclear space extended. Mitochondria are swollen with an enlightened matrix. In some cells there is a focal destruction of the crests, fragmentation and destruction of the internal membrane of mitochondria. GA tanks are vacually expanded. In many endothelial cells, fragmentation of the RER membranes and a sharp decrease in the number of ribosomes associated with its wall are observed.
As a result of edema of endothelial cells and aggregation of blood cells, the lumen of many hemocapillaries is sharply narrowed or closed. Basement membrane of hemocapillaries lengthened and thickened considerably showing fuzzy contours. In separate endothelial cells on the luminal surface of plasmollem there are sail-like protrusions (Fig. 2). Sometimes in hemocapillaries there are marked areas of lysis of luminal plasmollem, which is accompanied by the release of intracellular contents into the lumen of the microvessel. The submicroscopic analysis performed in 48 h after the start of the experiment, showed that the edema phenomenon in the endothelial cells continues to be maintained, but to a lesser extent expressed in comparison with the previous term of the study. Endothelial cell nuclei with low electron-density matrix, and marginal aggregation of chromatin granules is observed. Mitochondria are enlarged in volume with single disorientated ridges. Along with the expanded elements of the GA, fragmentation of the RER membranes is noted. Basement membrane is uneven, the alternation of thickened and narrow sections of it is revealed.
With the extension of the study (72 h), the alterations in the structural organization of hemocapillaries are more local in nature. Round-shaped nuclei with fine-grained nucleoplasma are seen. The nuclear membrane has winding contours and forms profound invaginations. Field around the nuclear space is expanded. Mitochondria are swollen, crests lose their parallelism, number of it is reduced. GA is represented by expanded tanks, vacuoles and bubbles. RER channels are expanded, filled with low electron-optical density content. In some cells the fragmentation of the RER membranes is noted, the number of ribosomes on the membranes of RER is reduced. Basement membrane locally thickened. The peripheral zone of the endothelial cells is sometimes enlarged, and sometimes sharply thinned with a significantly reduced number of mikropinocytosic vacuoles. In the lumen of hemocapillaries, adhesions, aggregation of leukocytes and aggregates of erythrocytes are found (Fig. 3). However, endothelial cells with signs of high functional activity are noted. In the cytoplasm of such cells, mitochondria of small size with a matrix of moderate electron-optical density are detected. The components of the GA are slightly enlarged. RER tanks are hypertrophied, rich in ribosomes. The performed experiments showed that after 12 h after the modeling of acute pancreatitis in the lungs there is a violation of hemomicrocirculation, as evidenced by the increased number of neutrophils in the lumen of the blood capillaries, their aggregation and adhesion to endothelial cells. Several other scientists point out the regional sequestration of neutrophils in the lungs, their adhesion and aggregation in acute lesions [6, 7]. Adhesion of neutrophils to the endothelium is the cause of the formation and release of oxygen radicals, as well as secretory degranulation. In this case, proteases (cathepsins, collagenase, elastase) of activated leukocytes damage not only the endothelium of hemocapillaries, but also the basal membrane.
Fig. 3. Erythrocyte aggregates in the lumen of the hemocapillary wall of the alveoli on 72 h of the experiment. Marking: 1 - erythrocyte; 2 - peripheral part of the endothelial cell; 3 -interstitial space; 4 - lumen of the alveoli. Electronic microphotography. Sat.: x6400.
Changes in a similar nature under various researchers [2, 12].
This is especially true for elactase [8, 10]. Destruction of granules of neutrophils, especially azurophilic ones, took place in our studies. With the continuation of the study period (24-72 h), an increase in the permeability of hemocapillaries is observed, which leads to the release of a part of the plasma and neutrophils in the interstitium and lumen of the alveoli with the development of interstitial and intraalveolar non-cardiogenic pulmonary edema. In this case, in hemocapillaries, along with the aggregation of leukocytes, there is an erythrocytic sludge and platelet aggregation, indicating a marked violation of hemomicrocirculation.
critical states are noted by a number of other
1. Studies have shown that acute L-arginine-induced pancreatitis is accompanied by pronounced changes in the submicroscopic structure of the hemomicrocirculatory bed of the lungs.
2. The nature and severity of ultrastructural changes in the hemomicrocirculatory bed of the lungs depends on the duration of the endogenous factor.
The prospect of further research: the study of the ultrastructural organization of other components of the aerohematic barrier in experimental acute pancreatitis is in the perspective of further research.
1. Avramenko AO, Smolyakov SM Patomorfologichni zminy u legeneviy tkanyni shchuriv cherez 6 and 24 hodyny pislya vnutrishnyoshlunkovoho vvedennya 2.5% vodnoho rozchinu amiaku. Klinichna ta Eksperimentalna Patologiya. 2016; 4 (58): 36. [in Ukranian]
2. Chernyayev AL Etiologiya, patogenez and patologicheskaya anatomy diffuznykh alveolarnykh porazheniy. Obshchaya reanimatologiya. 2005; 1 (5): 13-6. [in Russian]
3. Kassil VL Ostryi vnelegochnoy resoiratornyi sindrom distressa: opredeleniye, etiopatogenes, klinicheskiye i laboratornyie proyavleniya (obzor literatury c elementami kritiki). Klinicheskaya onkogematologiya. 2011; 4 (1): 54-65. [in Russian]
4. Kopichak IR Morfofunktsionalni zminy u lehenyakh z izolyovanymy ta kombinovanymy urazhennyamy. Klinichna khirurhiya. 2014; 1: 36-39. [in Ukranian]
5. Novikov NU, Tyshkevich LV, Jansuz KN Patomorphologicheskiye izmeneniya aerogematicheskogo barriera pri ostrom syndrome respiratornogo distressa v experimente. Tavricheskiy vestnik meditsiny i biologiyi. 2012; 15 (1): 169-175. [in Russian]
6. Orel YM Uchast neutrophilnykh granulotsytiv u rozvytku morphologichnykh zmin u lehenyakh pry yikh hostromu urazhenni v experymenti. Klinichna khirurhiya. 2015; 1: 43-5. [in Ukranian]
7. Teslyuk II. Krytychni stany: syndrome hostroho respiratornoho distressu. Terapiya. 2010; 11 (52): 41-3. [in Ukranian]
8. Wildman VN Ultrastructurnyie izmeneniya v respiratornykh kletkakh legkikh experimentalnykh zhyvotnykh s simulyatsiyei zakrytoy travmy grudi. Meditsina segodnya i zavtra. 2010; 2-3 (47-48): 36-44. [in Russian]
9. Cruz-Santamaria DM, Taxonera C, Iner M Update on pathogenesis and clinical management of acute pancreatitis. World J Gastrointest. Pathophisiol. 2012; 3(3):60-70.
10. Matthay MA, Zimmerman GA Acute lung injury and the acute respiratory distress syndrome: four decades of inquiry into pathogenesis and rational management. Am. J. Respir. Cell Mol.Biol. 2005; 33(4): 319-27.
11. Singh VK, Bollen TL, Wu BU, Kathryn Repas, Rie Maurer, Song Yu et al. An assessment of the severity of interstitial pancreatitis. Clin. Gastroenterol. Hepatol. 2011; 9:1098-103.
12. Williams AE The mercurial nature of neutrophils: still an enigma in ARDS? American J. of Physiology - Lung Cellular and Molecular Physiology. 2014; 306(3): 217-30.
13. Xi-Ping Zhang, JieZhang, Mei-Li Ma, Cai Y, Xu RJ, Xie Q, et.al Pathological changes at early stage of multiple organ injury in a rat model of severe acute pancreatitis. Hepatobiliary Pancreat Dis Int. 2010; 9(1):83-7.
14. Yap SC, Lee HT Acute kidney injury and extrarenal organ dysfunction. Anesthesiology. 2012; 166(5): 1139-48.
УЛЬТРАСТРУКТУРШ ЗМ1НИ ГЕМОКАП1ЛЯР1В ЛЕГЕНЬ ПРИ ЕКСПЕРИМЕНТАЛЬНОМУ ГОСТРОМУ ПАНКРЕАТИТ Заяць Л.М., Черкасова В.В.
У дослщах на 70 бших щурах-самцях лши Вютар електронномшроскошчним методом вивчено в динашщ (12, 24, 48, 72 год.) ультраструктурш змши гемомшроциркуляторного русла легень при експериментальному гострому панкреатит! Встановлено,
УЛЬТРАСТРУКТУРНЫЕ ИЗМЕНЕНИЯ ГЕМОКАПИЛЛЯРОВ ЛЕГКИХ ПРИ ЭКСПЕРИМЕНТАЛЬНОМ ОСТРОМ ПАНКРЕАТИТЕ Заяц Л.М., Черкасова В.В.
В опытах на 70 белых крысах-самцах линии Вистар электронномикроскопическим методом изучено в динамике (12, 24, 48, 72 ч.) ультраструктурные изменения гемомикроциркуляторного русла легких при
экспериментальном остром панкреатите. Установлено, что
що через 12 год. тсля початку дослщження спостертаються порушення субмкроскошчно! будови гемомкроциркуляторного русла легень. У гемокапiлярах альвеолярно! сттки вiдмiчаeться надмiрна акумуляцiя лейкоцита, !х адгезiя та агрегацiя. Зi збiльшенням термiну дослiдження (24-72 год.) у гемомкроциркуляторному руслi легень визначаються як дистрофiчно-деструктивш, так i компенсаторно-пристосувальш змiни.
Ключовi слова: легеш, гемомщоциркуляторне русло, гострий експериментальний панкреатит.
Стаття надшшла 4.01.18р.
через 12 ч. после начала исследования наблюдаются нарушения субмикроскопического строения гемомикроциркуляторного русла легких. В гемокапилярах альвеолярной стенки отмечается чрезмерная аккумуляция лейкоцитов, их адгезия и агрегация. С увеличением срока исследования (24-72 ч.) в гемомикроциркуляторного русле легких определяются как дистрофически-деструктивные, так и компенсаторно-приспособительные изменения.
Ключевые слова: легкие, гемомикроциркуляторное русло, экспериментальный острый панкреатит.
Рецензент Срошенко Г.А.
DOI 10.26724 / 2079-8334-2018-2-64-145-147 УДК 611.6+611.018+616-092.9
МОРФОЛОГ1ЧНА ХАРАКТЕРИСТИКА НИРКОВИХ Т1ЛЕЦЬ У ЩУР1В
E-mail: [email protected]
Метою дано! роботи було вивчення морфолопчних особливостей ниркових тшець в штактних щурiв та у групах контролю. Дослщження були проведенi на нирках взятих вщ 32 щурiв лшп Вiстар. Першу групу склали 5 iнтактних тварин, другу групу (контроль) - 9 тварин, яким вводили внутршньоочеревинно 1 мл фiзiологiчного розчину, третю групу (контроль) - 9 тварин, яким проводили розрiз шюри на зовнiшнiй поверхнi стегна з формуванням тдшюрно'! кишенi з подальшим ушиванням, четверту групу (контроль) - 9 тварин, яким вводили внутршньоочеревинно 1 мл фiзiологiчного розчину та робили розрiз на зовншнш поверхнi стегна з формуванням тдшюрно! кишет, з подальшим ушиванням рани. При дослщжент мкроскотчно! оргатзаци юрково! речовини нирок щурiв груп контролю, показали, що нирков1 тiльця збертають притаманну !м структурну органiзацiю, не викликае деструктивних змiн у структурах нефрона.
Ключовi слова: нирка, ниркове тiльце, щури..........................
Робота е фрагментом НДР "Експериментальнo-мoрфoлoгiчне вивчення ди mpaHcmaHmamie ^оконсервованог плаценти та шших екзогенних чиннитв на морфофункщональний стан внутрштх оргатв", № державноi реестраци 0113U006185.
Одшею з актуальних проблем сучасно! морфологи е вивчення будови нирок, яю займаються виведенням ¡з оргашзму иродукпв обм1ну речовин, регулящею водного балансу i юлькосп та складу бiльшостi електролiтiв, що забезпечують стабiльнiсть сольового гомеостазу i кислотно-лужно! рiвноваги [1, 2, 4, 5]. В науковш лiтературi даш про структурну органiзацiю та морфолопчш особливостi будови нирки iнтактних тварин численш, але ця тема до останнього часу залшаеться предметом уваги багатьох дослщниюв [2, 3, 5, 6]. Не останню роль вщграе розумiння особливостей структурно-функцiональних ироявiв з боку фшьтрацшного бар'еру нирок при рiзноманiтних станах [2, 5].
Мета дано! роботи полягае у вивченш морфолопчних особливостей ниркових тшець в штактних щурiв та у групах контролю.
Матер1али i методи дослщження. Дослiдження були ироведенi на нирках взятих вщ 32 щурiв лшп Вiстар. Забiр бiологiчного матерiалу для проведення дослiджень ироводився згiдно з "Правилами використання лабораторних ексиериментальних тварин" (2006, додаток 4) i Гельсшською декларацiею про гуманне вiдношення до тварин. Вщразу иiсля видалення нирки вирiзали iз середньо! частини юрково! речовини шматочки для мiкроскоиiчного дослщження. Першу груиу склали 5 штактних тварин, другу групу (контроль) - 9 тварин, яким вводили внутршньоочеревинно 1 мл фiзiологiчного розчину, третю груиу (контроль) - 9 тварин, яким проводили розрiз шюри на зовншнш поверхш стегна з формуванням тдшюрно! кишеш з подальшим ушиванням, четверту груиу (контроль) - 9 тварин, яким вводили внутршньоочеревинно 1 мл фiзiологiчного розчину та робили розрiз на зовншнш поверхш стегна з формуванням тдшюрно! кишет, з подальшим ушиванням рани. Матерiал для св^лово! мшроскопи компоненпв нирки готували зпдно iз загальноприйнятими правилами. Морфометричнии аналiз проводили за доиомогою нашвавтоматичного пристрою обробки графiчних зображень. Отриман цифровi данi обробляли методом варiацiиHоi ' статистики з використанням 1>критерда Ст'юдента.
Результати дослiдження та ix обговорення. Проведенi гiстологiчнi дослiдження
© А.А. Каиустянська, Ю.Б. Чайковський, 2018
