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9. Очеретнюк А. О. Гютолопчш змши в легенях щурiв протягом 7 дiб пiсля oniKy шюри Ш-А ступеня, площею 21-23 % поверхш тша та 1х кoрекцiя iзoтoнiчним розчином / А. О. Очеретнюк // Вюник морфологи. - 2012. - Т. 18, № 2. - С. 237-241.
10. Перов Ю. Л. Микроскопическая техника: руководство / Ю. Л. Перов // - М.: Медицина, -1996. - 544 с.
11. Сухомлин Т. А. Морфолопчш змши в легенях щyрiв при ошковш хвoрoбi та ix корекщя препаратом "Лшш" / Т. А. Сухомлин, Л. Г. Нетюхайло, Д. G. Нжоленко // Вюник проблем бюлоги i медицини. - 2014. - Т. 3, № 3 (112). - С. 196-199.
12. Bala S. An autopsy study of morphological changes of lung in burn patients with duration of hospital stay and total body surface area burned / S. Bala, S. Das, D. Guha [et al.] // International Journal of Research in Health Sciences. - 2014. - Vol. 2, № 2. - Р. 494-500.
13. Pham T. N. Risk factors for the development of pneumonia in older adults with burn injury / T. N. Pham, C. B. Kramer, M. B. Klein // J. Burn Care Res. - 2010. - Vol. 31, № 1. - Р. 105-115.
ОСОБЕННОСТИ СТРУКТУРНОЙ И МОРФОМЕТРИЧЕСКОЙ РЕОРГАНИЗАЦИИ СОСУДОВ ЛЕГКИХ ПРИ ЭКСПЕРИМЕНТАЛЬНОЙ ТЕРМИЧЕСКОЙ ТРАВМЕ Небесная З. М., Волков К. С., Литвинюк С. О.
В эксперименте на половозрелых белых крысах-самцах проведены гистологические и морфометрические исследования легких после термической травмы. Установлено, что степень структурных и морфометрических изменений сосудов органа зависит от срока исследования. В ранние сроки после ожогового повреждения (1-7 сутки эксперимента) установлены приспособительно-
компенсаторные изменения и начальные признаки деструкции сосудистых компонентов органа. В поздние сроки (14-21 сутки эксперимента) обнаружены значительные деструктивно-дегенеративные сосудистые расстройства.
Ключевые слова: сосуды легких, гистологические и морфометрические изменения, термическая травма.
Стаття надшшла 28. 08. 2015 р.
PECULIARITIES OF STRUCTURAL AND MORPHOMETRIC REORGANIZATION OF THE LUNGS VESSELS AFTER EXPERIMENTAL
THERMAL TRAUMA Nebesna Z. M., Volkov K. S., Lytvynyuk S. O.
In the experiment on mature white male rats histological and morphometric research of the lungs were made after thermal injury. It was established that the degree of structural and morphometric vascular changes in components of organ depends on the term of experiment. In the early stages after burn injury (1-7 days of experiment ) by adaptative-compensatory changes and initial signs of destructive of vascular in components of organ were revealed. In the later stages (14-21 days of experiment) significant destructive-degenerative vascular disorders were found.
Key words: pulmonary vessels, histologic and morphometric changes, thermal trauma.
Рецензент Срошенко Г.А.
UDC 611.41:615.348:599.323.4
MICROMORPHOMETRIC CHANGES IN RATS' SPLEEN IN THE FIRST 7 DAYS AFTER SKIN BURNS AND UNDER APPLICATION HYPEROSMOLAR COLLOID SOLUTIONS
In this work on the laboratory white male rats weighing 155-160 g studied stereometric relative volume changes of white and red pulp of the spleen under the influence of drugs infusion 0.9% solution of NaCl, lactoproteins with sorbitol and HAES-LX-5% after 1, 3 and 7 days without burns and after burns skin damage II-III degree with affected area 21-23% of body surface. Found no change of data stereometric parameters when injected to animals without skin burns during the first seven days these solutions. When using hyperosmolar colloid infusion solutions lactoproteinum with sorbitol and HAES-LX-5% from the third day after burn of skin there is a pronounced positive correction changes of relative volume of white and red pulp of the spleen, compared with the use of 0.9% solution of NaCl.
Keywords: spleen, rats, stereometry, skin burns, colloidal hyperosmolar solutions.
The study was a fragment of SRW "Structural changes in the lungs in conditions of endogenous intoxication that caused by skin burns and its correction by domestic infusion drugs lactoproteinum with sorbitol and HAES-LX-5% (experimental research)" (№ state registration: 0112U004187).
Burn shock is one of the most frequent causes of death in burn disease. It can appear on condition of inadequate assistance in the early stages after injury [8]. Therefore, actively pursued the development of modern drugs that normalize blood electrolyte composition in conditions of burn shock. These domestic drugs are lactoproteins with sorbitol - serially produced by Kiev "Biopharma" and HAES-LX-5% -developed in the SI "Institute of Pathology of Blood and Transfusion Medicine Medical Sciences of Ukraine" (Lviv) [10]. On the reduction of endogenous intoxication in rats after burn of skin 2-3 level, 2123% of the surface area of the body and hepatoprotective effect at correction of infusion solutions lactoproteinum with sorbitol and HAES-LX-5 in terms of experimental burn disease in the early stages (1, 3 and 7 days) indicate studies conducted in the Vinnitsa National Medical University named after Pirogov [6, 7, 11]. However, research of action infusion drugs lactoproteins with sorbitol and HAES-LX-5% on the structure of spleen both intact rats and rats in the early stages (1, 3 and 7 days) after the burn of skin is not conducted.
Objective - explore stereometric changes in spleen in rats at 7 days after the burn skin and entering in first seven days of the experiment 0.9% solution of NaCl, lactoproteinum with sorbitol or HAES-LX-5%.
Materials and methods. Studies were performed on laboratory white male rats weighing 155160 g, obtained from the vivarium SI "Institute of Pharmacology and Toxicology AMS of Ukraine." During the experiment, all animals kept in vivarium conditions VNMU named after Pirogov (room temperature - in the range 24-25 ° C, humidity - in the range 40-60%) on a standard water-diet, with free access to food and water. All experiments were carried out taking into account recommendations the European Commission to conduct medical and biological research using animals and medical recommendations, State Pharmacological Center MoH Ukraine and "Rules for clinical safety evaluation of pharmacological agents (GLP)" and rules for the humane treatment of experimental animals (approved by the Committee of bioethics VNMU named after Pirogov - protocol № 1 from 14.01.2010).
All solutions were injected into the inferior vena cava after aseptically catheterization through the femoral vein at a dose of 10 ml / kg body weight - average therapeutic dose was calculated by developers HAES-LX-5% according to existing recommendations [12].
Burn skin damage caused by applying to previously shaved rat lateral surfaces of the body for 10 seconds by four copper plates (two plates on each side, each with a surface area on 13.86 cm2) that previously within 6 minutes heated in water at a constant temperature 100 C° [13, 15]. Thus, according to the formula M.O. Lee [14] the total area of the skin destruction in rats was 21-23%. This area at a given exposure is sufficient to form burns II-III degree (according to the classification adopted at the 20 th Congress of Surgeons of Ukraine, Ternopil, 2000) and induce shock of medium severity [8, 9].
Shaving of animals, staging burns, major vessels catheterization and decapitation of animals carried out under conditions of intravenous (60 mg / kg) propofol anesthesia.
Samples for histological examination of the spleen were fixed in 10% neutral formalin solution, washed in running water, left the water in the battery alcoholic solutions of increasing concentration and included in paroplast [2, 4]. 4-6 micron thick sections were made on a rotary microtome, placed on a piece of subject glass after standard hematoxylin eosin stained posting and embedded in Canada balsam. Histological preparations examined in the light microscope OLYMPUS BH-2 using lenses x10 and x40 and eyepiece x10.
Stereological spleen tissues research conducted at the demonstration screen of microscope Laborlux S (Leitz) with an increase 40/1,25x10 using nets of Wejbel [3]. For the following formula was determined volume density (relative volume, cm3/cm3) red and white pulp of the spleen: Vvi = Pi / PT, where Vvi - volume density of corresponding sections of the spleen; Pi - the number of test points, attributable to the relevant structures; PT - the total number of test points. At each histological preparation (6 animals in each group) in different areas of the spleen were selected by random selection for 5 visual fields that were evaluated by stereological parameters.
Statistical analysis of the results was conducted in the package "Statistica 6.1" (belongs to SRC VNMU named after Pirogov, license № BXXR901E246022FA) using nonparametric methods for assessing the results.
Results and discussion. Dynamics of changes in the relative volume of white and red pulp of the spleen of rats after entering in the first seven days of the experiment 0.9% solution of NaCl, Lactoproteinum with sorbitol or HAES-LX-5%, and in animals after skin burns and similar entering listed infusion solutions presented in tables 1 and 2.
Between groups of animals without burn of the skin, which during the first seven days of the experiment were injected 0.9% solution of NaCl, Lactoproteinum with sorbitol or HAES-LX-5% does not have significant differences or trends relative volume of white and red pulp of the spleen after 1, 3 and 7 days of the experiment (Table. 1, 2). Attract attention only larger by 10-13% (p> 0.05) value of the relative volume of white pulp and lower at 11.7% (p> 0.05) the relative amount of red pulp in rats after entering of lactoproteins sorbitol than in animals injected 0.9% NaCl solution or HAES-LX-5%.
Between groups of animals after burn your skin, which during the first seven days of the experiment injected thesame infusion solutions in the same dose also not found significant differences of relative volume of white and red pulp of the spleen after 1, 3 and 7 days of the experiment (see. Table. 1, 2). Established only following trends: to lower (by 20.1%) values (p = 0.056) of relative volume of white pulp in 7 days after burn of skin when injected 0.9% solution of NaCl, compared with a similar group after 1 day after the burn skin; to higher (by 27.7%) values (p = 0.060) of relative volume of white pulp in 7 days after burn of skin when injected with sorbitol solution Lactoproteinum compared to rats that injected after skin burn 0.9% solution of NaCl. Also draws attention following changes in relative volume of white and red
pulp in animals after burn of skin on a background of the introduction of colloidal hyperosmolar solution (see. Table. 1, 2): reduction when injected Lactoproteinum with sorbitol by 4.1% (p> 0, 05), and when injected HAES-LX-5% by 7.8% (p> 0.05) relative volume of white pulp from 1 to 7 days of the experiment; larger (by 14.4 and 18.0%, p> 0.05) value of the relative volume of white pulp and lower (by 7.1 and 9.1%, p> 0.05) value of the relative volume of red pulp in rats in 3 days after burn of skin when injected HAES-LX-5% or lactoproteins with sorbitol than in animals at this time when administered 0.9% NaCl solution; larger by 19.4% (p> 0.05) value of the relative volume of white pulp and lower by 5.8% (p> 0.05) value of the relative amount of red pulp in rats in 7 days after burn of skin when entering solution HAES-LX-5% than the animals in this term when injected 0.9% solution of NaCl; lower by 7.9% (p> 0.05) value of the relative amount of red pulp in rats in 7 days after burn of skin when injected sorbitol solution with Lactoproteinum than in animals in the term when injected 0.9% NaCl solution.
Table 1
Changes in relative volume of white pulp of the spleen in rats during 7 days after the burn of skin
Ivinn pnllmHal (ivn^rncmnlar cnlnfinn inrn3 / rm3, 1M4-
0,9% NaCl Lactoproteinum with sorbitol HAES-LX-5% P1 P2 P3
1 day 0,398±0,131 1 day 0,440±0,151 1 day 0,403±0,124 >0,05 >0,05 >0,05
3 day 0,401±0,128 ** 3 day 0,448±0,131 ** 3 day 0,416±0,108 * >0,05 >0,05 >0,05
7 day 0,405±0,156 ** 7 day 0,460±0,165 * 7 day 0,413±0,156 t >0,05 >0,05 >0,05
P1-3 >0,05 P1-3 >0,05 P1-3 >0,05
P1-7 >0,05 P1-7 >0,05 P1-7 >0,05
P3-7 >0,05 P3-7 >0,05 P3-7 >0,05
Burn + 0,9% NaCl Burn + Lactoproteinum with sorbitol Burn + HAES-LX-5% P1 P2 P3
1 day 0,348±0,122 1 day 0,370±0,140 1 day 0,360±0,171 >0,05 >0,05 >0,05
3 day 0,305±0,169 3 day 0,360±0,143 3 day 0,349±0,151 >0,05 >0,05 >0,05
7 day 0,278±0,127 7 day 0,355±0,148 7 day 0,332±0,108 =0,060 >0,05 >0,05
P1-3 >0,05 P1-3 >0,05 P1-3 >0,05
P1-7 =0,056 P1-7 >0,05 P1-7 >0,05
P3-7 >0,05 P3-7 >0,05 P3-7 >0,05
Notes: here and in the future 1. p1 - significant differences between groups of 0.9% NaCl solution and Lactoproteinum with sorbitol without burn or with burn of skin. 2. p2 - significant differences between groups of 0.9% NaCl solution and HAES-LX-5% without burn or with burn of skin. 3. p3 - significant differences between groups Lactoproteinum with sorbitol and HAES-LX-5% without burn or with burn of skin. 4. pi_3 -significance of differences within the respective groups between the 1and 3 day experiment. 5. p1-7 - significance of differences within the respective groups between the 1and 7 day experiment. 6. p3.7 - significance of differences within the respective groups between the 3and 7 day experiment. 7. *, **, *** - significance of differences (in level p> 0,05 - *, p> 0,01 - ** and p> 0,001 - ***) within the respective groups and the corresponding terms between rats without burn and with skin burns (marked larger value). 8. t - trends differences within the respective groups and the corresponding terms between rats without burn and with skin burns (marked larger value).
Table 2
Changes in relative amount of red pulp of the spleen in rats during 7 days after the burn of skin and
33
0,9% NaCl Lactoproteinum with sorbitol HAES-LX-5% P1 P2 P3
1 day 0,600±0,132 1 day 0,558±0,151 1 day 0,597±0,124 >0,05 >0,05 >0,05
3 day 0,599±0,128 3 day 0,548±0,128 3 day 0,584±0,108 >0,05 >0,05 >0,05
7 day 0,594±0,156 7 day 0,534±0,161 7 day 0,571±0,148 >0,05 >0,05 >0,05
P1-3 >0,05 P1-3 >0,05 P1-3 >0,05
P1-7 >0,05 P1-7 >0,05 P1-7 >0,05
P3-7 >0,05 P3-7 >0,05 P3-7 >0,05
Burn + 0,9% NaCl Burn + Lactoproteinum with sorbitol Burn + HAES-LX-5% P1 P2 P3
1 day 0,647±0,123 1 day 0,630±0,140 t 1 day 0,640±0,171 >0,05 >0,05 >0,05
3 day 0,693±0,167 * 3 day 0,637±0,142 ** 3 day 0,644±0,144 t >0,05 >0,05 >0,05
7 day 0,699±0,133 * 7 day 0,644±0,148 ** 7 day 0,659±0,139 * >0,05 >0,05 >0,05
P1-3 >0,05 P1-3 >0,05 P1-3 >0,05
P1-7 >0,05 P1-7 >0,05 P1-7 >0,05
P3-7 >0,05 P3-7 >0,05 P3-7 >0,05
Between groups of animals without burn and after burn of the skin, which during the first seven days of the experiment injected hyperosmolar colloid infusion solutions established the following trends and significant differences in relative volume of white and red pulp of the spleen after 1, 3 and 7 days of the experiment (Table . 1, 2): significantly larger (p <0,05-0,01) and a small trend (p = 0.063) values relative to larger volume of white pulp in animals without skin burn that obtained 0.9% solution of NaCl (by 23.9% after 3 days and by 31.6% after 7 days) Lactoproteinum with sorbitol (by 19.6% after 3 days and by 22.8% after 7 days) or HAES-LX-5% (by 16.1% after 3 days and at 19.6% after 7 days) than in similar groups of
rats after burn of skin, and vice versa significantly lower (p <0,05-0,01) and minor trend (p = 0,060-0,068) to smaller values of relative amount of red pulp in animals without skin burn that obtained 0.9% solution of NaCl (at 15.7% after 3 days and by 17.7% after 7 days) Lactoproteinum with sorbitol (by 12.9% after 1 day, by 16.2% after 3 days and by 20.6% after 7 days) or HAES-LX-5% (by 10.3% after 3 days and by 15.4% after 7 days) than in similar groups of rats after burn of skin.
That is, using hyperosmolar colloid infusion solutions Lactoproteinum with sorbitol and HAES-LX-5% from the third day after burn of skin observed positive correction of changes of relative volume of white and red pulp of the spleen, compared to using 0.9% NaCl solution.
It should be noted that changes in relative volume of white and red pulp in rats without burn and after burn of the skin, which during the first seven days of the experiment injected colloidal hyperosmolar solutions coincide with the peculiarities of the structural changes in the spleen of rats on set by us lightoptical level [1, 5].
1. When entering rats during the first seven days hyperosmolar colloid solutions are not found significant changes in value relative volume of white and red pulp.
2. After the skin burn injury when injected 0.9% NaCl solution relative volume of white pulp after 3 days by 23.9% lower (p <0.01), and after 7 days - by 31.6% lower (p <0 01) than in rats without the skin burn when injected this solution in similar terms of observation; and the relative amount of red pulp contrary, after 3 days by 15.7% greater (p <0.05) and after 7 days - by 17.7% greater (p <0.05).
3. After the skin burn injury when injected sorbitol solution with Lactoproteinum relative volume of white pulp after 3 days by 19.6% lower (p <0.01), and after 7 days - by 22.8% lower (p <0.05) than in rats without the skin burn when injected this solution in similar terms of observation; and the relative amount of red pulp contrary, after 3 days by 16.2% greater (p <0.01) and after 7 days - by 20.6% greater (p <0.01).
4. After the skin burn injuries when injected solution HAES-LX-5% relative volume of white pulp after 3 days by 16.1% lower (p <0.05), and after 7 days - by 19.6% lower (p = 0.067) than in rats without the skin burn when injected this solution in similar terms of observation; and the relative amount of red pulp contrary, after 3 days by 10.3% greater (p = 0.050) and after 7 days - by 15.4% greater (p <0.05).
Prospects for further research - results obtained proved the possibility of using HAES-LX-5% at a dose of 10 ml per kg in subsequent studies.
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5. Huminskyy Yu.Y. Dynamics of morphological changes in the spleen of rats in early stage after burn of skin 2-3 degrees 2123% area of the skin and their correction by infusion of solution HAES-LX-5% / Yu.Y. Huminskyy, N.P. Ocheretna // Biomedical and Biosocial Anthropology. - 2013. - № 21. - P. 93-97. (in Ukrainian)
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М1КРОМОРФОМЕТРИЧН1 ЗМ1НИ В СЕЛЕЗ1НЦ1 ЩУР1В У ПЕРШ1 7 Д1Б П1СЛЯ ОП1КУ ШК1РИ ТА ПРИ ВВЕДЕНН1 КОЛО1ДНИХ Г1ПЕРОСМОЛЯРНИХ РОЗЧИН1В Очеретна Н.П.
В робой, на лабораторних бших щурах-самцях масою 155-160 г вивчеш стереометричш змши вщносного обему бшо! i червоно! пульпи селезшки при дп iнфузiйних препаратiв 0,9% розчину NaCl, лактопротеiну з сорб^олом та HAES-LX-5% через 1, 3 i 7 дiб без отку та пiсля опiкового пошкодження шюри II-III ступеню з площею ураження 21-23 % поверхнi тша. Встановлено вiдсутнiсть змiн даних стереометричних показниюв при введеннi тваринам без отку шюри протягом перших семи дiб даних розчинiв. При використанш iнфузiйних коло'дних гiперосмо-лярних розчинiв лактопроте'ну з сорбiтолом та HAES-LX-5% починаючи з третьо! доби пiсля отку шюри спостертаеться виражена позитивна корекщя змiн вiдносного об'ему бiлоi' i червоно! пульпи селезiнки, порiвняно iз застосуванням 0,9 % розчину NaCl.
Ключовi слова: селезiнка, щури, стереометрiя, опiк шкiри, коло'дш гiперосмолярнi розчини.
Стаття надшшла 28.08.2015 р.
МИКРОМОРФОМЕТРИЧЕСКИЕ ИЗМЕНЕНИЯ В СЕЛЕЗЁНКЕ КРЫС В ПЕРВЫЕ 7 ДНЕЙ ПОСЛЕ ОЖОГА КОЖИ И ПРИ ВВЕДЕНИИ КОЛОИДНЫХ ГИПЕРОСМОЛЯРНЫХ РАСТВОРОВ Очеретная Н.П В работе, на лабораторных белых крысах-самцах массой 155-160 г изучены стереометрические изменения относительного объема белой и красной пульпы селезёнки при действии инфузионных препаратов 0,9% раствора NaCl, лактопротеина с сорбитолом и HAES-LX-5% через 1, 3 i 7 дней без ожога и после ожогового повреждения кожи II-III степени с площадью поражения 21-23 % поверхности тела. Установлено отсутствие изменений данных стереометрических показателей при введении животным без ожога кожи на протяжении первых семи дней данных растворов. При использовании инфузионных коллоидных гиперосмолярных растворов лактопротеина с сорбитолом и HAES-LX-5%, начиная с третьих суток после ожога кожи, наблюдается выраженная позитивная коррекция изменений относительного объёма белой и красной пульпы селезёнки, в сравнении с использованием 0,9 % раствора NaCl.
Ключевые слова: селезёнка, крысы, стереометрия, ожог кожи, коллоидные гиперосмолярные растворы.
Рецензент Гунас В.В.
УДК 577.17:591.436:599.323.4
1НСУЛ1НОПОД1БНИЙ ФАКТОР РОСТУ-1 У ПАТОГЕНЕЗ1 ХРОН1ЧНОГО АЛКОГОЛЬНОГО УШКОДЖЕННЯ ПЕЧ1НКИ ЩУР1В Р1ЗНИХ В1КОВИХ ГРУП
В останне десятирiччя, хрошчш дифузш хвороби печшки через високий рiвень захворюваност та небезпечш для життя наслщки вiдносяться до актуальних проблем сучасно! медицини. Актуальним питанням е дослщження молекулярних механiзмiв репаративно! регенерацп патологiчно змiненоi' печiнки та впливiв на данi процеси медикаментiв. Перспективним являеться вивчення рiвня iнсулiноподiбного фактора росту-1 (IGF-1) у щурiв рiзного вiку при хронiчному алкогольному ушкодженш печiнки та за умов фармакокорекцп кверцетином та L-аргiнiном Ь-глутаматом, а також дослщження кореляцiйних взаемозв'язюв мiж IGF-1 та змiнами бiохiмiчних показникiв сироватки кровi. Доведено, що при хронiчному алкогольному ураженнi печшки вщбуваеться зменшення концентрацп IGF-1 у тварин уах вiкових груп, яке корелюе iз бiохiмiчними маркерами синдромiв цитолiзу та холестазу. Встановлено, що за умов фармакокорекцп Ь-аргшшом Ь-глутаматом достовiрно збiльшуеться рiвень IGF-1, при цьому наявш прямi достовiрнi кореляцшш зв'язки з показниками функцiонального стану печшки, що вказуе на вщновлення структури паренхiми печiнки та !! функцiонального стану за рахунок активацп репаративно! регенерацп. Одержанi данi свщчать, що гепатопротектор L-аргiнiн Ь-глутамат володiе бiльш позитивним впливом на процеси репаративно! регенерацп у порiвняннi iз кверцетином.
Ключовi слова: вiковi особливоси, хронiчне алкогольне ушкодження печiнки, iнсулiноподiбний фактор росту-1, бюх1м1чш показники.
Робота е фрагментом НДР «Вiковi особливостi патогенезу гостроI i хрошчноХ патологи внутрштх оргатв. Патогенетичт тдходи до л^вання» (номер державноI реестраци 011Ш008679).
В останш роки, алкогол1зм, в усьому свт 1, зокрема, в Укра'ш характеризуеться значним поширенням та омолодженням (високий темп росту шдл^ково! алкогол1зац1!). Так, тенденщя до зменшення в1ку ос1б, яю вживають алкоголь, фем1н1зац1я (все бшьша к1льк1сть д1вчаток-п1дл1тк1в починае вживати алкогольш напо!) та потенц1йно важк1 наслщки викликають значне занепокоення [18]. Експерти ВООЗ вважають ситуащю «небезпечною», якщо доза алкоголю на одну людину
