facebook
twitter
vk
instagram
linkedin
google+
tumblr
akademia
youtube
skype
mendeley
Wiki
Global international scientific
analytical project
GISAP
GISAP logotip

SOME TISSUE METABOLISM MARKERS CONTENT IN BLOOD SERUM OF THE IMMATURE RATS UNDER INFLUENCE OF GRAVITATIONAL OVERLOADS AND GLUTARGIN USE

Автор Доклада: 
Moroz G.A., Kutya S.A., Pikalyuk V.S., Kriventsov M.A.
Награда: 
SOME TISSUE METABOLISM MARKERS CONTENT IN BLOOD SERUM OF THE IMMATURE RATS UNDER INFLUENCE OF GRAVITATIONAL OVERLOADS AND GLUTARGIN USE

SOME TISSUE METABOLISM MARKERS CONTENT IN BLOOD SERUM OF THE IMMATURE RATS UNDER INFLUENCE OF GRAVITATIONAL OVERLOADS AND GLUTARGIN USE

 

 

 

Moroz Gennadiy Alexandrovich, candidate of medical science, associate professor
Kutya Sergey Anatolievich, candidate of medical science, associate professor
Pikalyuk Vasiliy Stepanovich, doctor of medical science, professor
Kriventsov Maxim Andrеevich, candidate of medical science, assistant
State Institution «Crimea State Medical University named after S.I. Georgievsky»

 

Article shows results of investigation of TBA-active products, potassium, calcium, phosphorus, acid and alkaline phosphatase blood serum content in immature rats exposed to hypergravity (9g, daily, 10 minutes, 10 and 30 days) with glutargin use. It is revealed, that systematic hypergravity caused disturbances in lipid, protein, electrolite metabolism. Lipid peroxidation was activated (TBA-active products content increased). Synthetic processes and bone matrix mineralization were altered (low alkaline phosphatase content), glycolysis was activated at early stages. Further hypoxia rise inhibited glycolysis result in changes of some cations blood serum content. Glutargin use normalizes serum blood level most of investigated parameters.
Keywords: blood serum, rats, hypergravity, glutargin

В работе представлены результаты исследования содержания ТБК-активных продуктов, калия, кальция, фосфора, кислой и щелочной фосфатаз в сыворотке крови неполовозрелых крыс, подвергавшихся воздействию гравитационных перегрузок (9g, ежедневно, 10 минут, 10 и 30 дней) на фоне парентерального введения глутаргина. Установлено, что при систематическом воздействии гипергравитации в организме происходят сдвиги липидного, белкового и электролитного обмена. При нарушении метаболизма липидов активируется их перекисное окисление, свидетельством чего является изменение содержания ТБК-АП. Изменяются синтетические процессы и темпы минерализации костного матрикса (уменьшение содержания ЩФ), на ранних стадиях гипоксии активируется гликолиз. При дальнейшем нарастании гипоксии наблюдается угнетение гликолитических процессов, нарушение электролитного баланса, что приводит к изменению содержания ряда катионов в сыворотке крови. Применение глутаргина способствует нормализации сывороточного уровня большинства изучавшихся показателей.
Ключевые слова: сыворотка крови, крыса, гипергравитация, глутаргин.


Normal organism functioning is provided by the internal environment constancy. Here, along with proteins, nucleic acids, lipids and carbohydrates, mineral substances also play important role, deficiency or excess of which causes different pathological conditions.

Most important minerals, required for normal functioning of the body, are calcium and phosphorus. In humans and animals calcium plays an important role in regulating the cell membranes permeability, muscular, nervous and glandular tissues electrogenesis, and secretory processes of exo- and endocrine glands. Calcium ions are involved in the activation of enzyme systems. Deviation of the calcium concentration in the blood and tissues versus normal level leads to the development not only functional, but also morphological disturbances in number of organs and systems [1]. Phosphorus is found in vitamins, coenzymes, various macroergic compounds; it is a structural component of DNA and RNA, providing protein synthesis, and involved in oxidative phosphorylation and ATP synthesis. Phosphorus is also an important component, ensuring functioning of both skeletal and cardiac muscle tissue. Together with calcium, as a component of the bone mineral structure - hydroxyapatite, they form the basis of solid bone matrix, while being in a state of chemical equilibrium with the calcium and phosphorus ions in serum [1]. Phosphatases - hydrolyzing enzymes, which catalyze cleavage of ester bonds in the monoester of phosphoric acid - play important role in the metabolism of carbohydrates, nucleotides and phospholipids. It is considered, that alkaline phosphatase (ALP) is involved in the extracellular breakage of pyrophosphate - protein inhibitor of calcium phosphate deposition. In addition, it is a plasma marker of the osteoblast activity [2]. Acid phosphatase (ACP) is an enzyme, which catalyzes hydrolysis of orthophosphoric monoesters with the phosphate group removal. This enzyme exhibits optimal activity in acidic environment. In macrophages and osteoclasts high activity of ACP is observed, which allows considering it, as a marker of a bone resorption [2]. In providing of the homeostasis potassium ions also play significant role. It is known, that potassium is involved in maintaining of the electric membrane potential of the cells and in the regulation of intracellular osmotic pressure. It stimulates activity of glycolytic enzymes and are involved in the protein metabolism, plays an important role in neural and muscle cells, participates in conduction of nerve impulses, has immunomodulatory activity [3, 4]. There is no pool of potassium in the body, so even small changes in the intracellular potassium concentration considerably modify its plasma concentration. Intake of potassium into cells is stimulated by insulin, catecholamines and aldosterone. One of the reasons for hypokalaemia is excessive adrenal response to stress [5, 6].

It is known, that metabolic and energy alterations under influence of the damaging factors, as a result of lipid peroxidation (LPO), lead to the accumulation of free radicals in tissues, which initiate cell damage with subsequent development of various pathological conditions. Products of free radical reactions (thiobarbituric acid-active products, TBA-AP) significantly affect catalytic processes in cells, structure and properties of cell membranes, as well as the nature of the regulatory effect manifestations of the various biologically active compounds, participating in the maintenance of homeostasis [7-9].

To date, any published information related to microelements, enzymes and LPO activity markers content in serum under stress factors conditions, such as the gravitational overloads, is absent. Above mentioned information, as well as necessity to develop new patterns to improve the resistance in response to hypergravity, have identified the research purpose: to study TBA-AP, potassium, calcium, phosphorus, ACP and ALP content in serum of rats with systematic exposure of significant magnitude gravitational overloads, as well as under pharmacological intervention with antioxidant drug - glutargin.

Materials and methods. Experiments were carried out on 48 immature male Wistar rats with initial weight 120-130 g. Rats were divided into the four series. Rats from first series were exposed to daily transverse gravitational accelerations (9G) in forms of consecutive three "platforms" with duration 3 minutes each (with two 30-second interruptions in between them). Hypergravity was simulated by rotating the animals at the peripheral containers at the centrifuge C-2/500 (operating range from 1 to 50 G, radius 50 cm, gradual onset rate - 1,6 G/s, gradual decline rate - 0,6-0,8 G/s). Rats, which for duration of the hypergravity session were placed in similar containers on a platform of centrifuge (Control-1), were used as controls for the first series. Second group of animals received glutargin intraperitoneally in a dose of 100 mg/kg body weight 30 minutes prior to the hypergravity session [10]. This drug is antioxidant, inhibiting lipid peroxidation at different stages; it participates in the maintenance of systemic and local hemodynamic parameters due to the nitric oxide formation from L- arginine L- glutamate (active ingredient). Rats, received intraperitoneally sterile saline solution in equivalent volumes 30 minutes prior to placing into containers, were used as control for this series (Control-2).

At the end of the experiment (after 10 and 30 days) rats were decapitated under ether anesthesia. Blood samples were removed with subsequent centrifugation and obtaining of serum. The serum calcium levels were measured by trilonometry titration in the presence of murexide, while potassium levels were measured by flame photometer “Corning-410” and content of phosphorus, ACP and ALP activities - by photoelectric colorimeter CPC-3. Analysis of the TBA-AP content was carried out using standard set of reagents "TBA-AGAT".

Data obtained from the study were processed using methods of variation statistics. In all cases, statistical signi?cance was inferred for p<0.05.

Results and discussion. 10-days of gravitational overloads caused decrease in the TBA-AP in rat’s serum with 7.74 % (p < 0.05) compared with control data (Fig. 1). Under 30 days exposure activation of lipid peroxidation processes was noted, which was confirmed by increasing TBA-AP content by an average of 11.46% (p < 0.05) (Table 1, Fig. 2).

Table 1

Potassium, calcium, phosphorus, ACP, ALP and TBA-AP content in the serum of immature rats under exposure to hypergravity and its correction by administration of glutargin (M ± m)

Changes in the content of certain compinents of blood serum of rats after 10-day exposure to hypergravity

Changes in the content of certain components of blood serum of rats after 30-day exposure to hypergravity

Determination of potassium serum levels in rats, systematically exposed to hypergravity, revealed a significant difference in comparison with control, content increasing under 10-day exposure - with 20.85% (p < 0.05), while under 30-day exposure - with 22.33% (p < 0.05) (Table 1, Fig. 1 and 2). This can be explained by the fact, that repeated exposure to gravitation overloads depleted catecholamines and mineralocorticoids reserves in the rat’s adrenal glands during first days of the stress and reduces their blood levels with compensational potassium leakage from the cells to the plasma.
Exposure to gravitation overloads similarly led to serum calcium level increase: under 10-day exposure - with 5.78% (p < 0.05), and under 30-day exposure - with 6.91% (p < 0.05) (Table 1, Fig. 1 and 2). Determination of phosphorus content revealed no significant differences from control values at both follow-up periods, although was noted a tendency to its increasing. In view of data limitation, concerning effect of gravitational acceleration on the major phosphorus - calcium metabolism regulators, the relevance of this finding has to be taken with caution. Possible reasons for hypercalcemia are increased release of calcium from bone tissue or reducing of its absorption, increased calcium absorption in the small intestine or reduced excretion of calcium in the kidneys.

Investigation of the activity of bone metabolism markers in the rat serum revealed decrease ALP activity with 9.50% (p < 0.05) under 10-day exposure, and with 16.29% (p<0.05) under 30-day exposure, which may indicate inhibition of bone formation or retardation of the bone tissue mineralization (Table 1, Fig. 1, 2). ACP activity was reduced in both observations, but the results were not statistically significant.

To stabilize the oxidative homeostasis in rats, exposed to 10-day gravitational overloads, glutargin administration did not show expected effect on lipid peroxidation, since the content of TBA-AP only insignificantly decreased versus control values with 8.68 % (p > 0.05) (Fig. 1). Under glutargin administration after 30 days of experiment TBA-AP content was within control values (Table 1, Fig. 2).

Both potassium and calcium blood levels at both follow-up periods were below the control values with 6.28% (р>0.05) and 6.35% (р>0.05), 1.46% (р>0.05) and 2.74% (р>0.05), respectively (Table 1, Fig. 1 and 2). Such decline may be related to the glutargin ability to stimulate renal blood flow [9], accompanied by acceleration of the elimination of these cations by the kidneys.

At the same time, administration of the glutargin under 10-day exposure to gravitational overloads contributed to normalization of the ACP and ALP decreased activities, demonstrating thereby protective properties. Under 30-day exposure glutargin stimulated activity of both studied phosphatases, whose content was increased in comparison with control: ACP – at 11.88% (p< 0.05) and ALP – at 10.98% (р>0.05) (Table 1, Fig. 2).

Conclusion Thus, based on the results obtained, it can be concluded that in rats with systematic exposure to hypergravity occurs variations of lipid, protein and electrolyte metabolisms, which are typical for respiratory and circulatory hypoxia, proceeding under the influence of gravitational overloads. In case of metabolic disturbance of lipids their peroxidation is activated, which is supported by the identified changes in the TBA-AP content. Intermediate products of protein metabolism are accumulated. Synthetic processes and rates of the bone matrix mineralization are changed (decrease in the ALP concentration). In the early stages of hypoxia there is activation in the glycolysis. With increasment of the hypoxia, there is a further development of electrolyte imbalance, which leads to significant changes in serum levels of potassium and calcium.

Administration of the antioxidant drug glutargin contributes to normalizing serum levels of the most of the studied parameters.

In perspective there are plans to investigate parallels between the identified changes and growth processes in the skeleton of rats, as well as the morphology and function of the adrenal glands under conditions of systematic exposure to gravitational overloads.

References:
1. Ковалев Д. И. Регуляция обмена кальция в организме человека / Д. И. Ковалев // Проблемы эндокринологии. – 1991. – Т. 37, № 6. – С. 61-66.
2. Остеопороз: эпидемиология, клиника, диагностика, профилактика и лечение [монография] / Н. А. Корж, В. В. Поворознюк, Н. В. Дедух, И. А. Зупанец. – Х.: Золотые страницы, 2002. – 650 с.
3. Абдрешов С. Н. Морфофункциональное состояние шейных лимфотических узлов в условиях микрогравитации / С. Н. Абдрешов, Л. Э. Булекбаева, Г. А. Демченко // Бюллетень СО РАМН. – 2008. – № 2 (130). – С. 30–34.
4. Айзман А. И. Регуляция гомеостаза калия: возрастные особенности / А. И. Айзман // Нефрология и диализ. – 2001. – Т. 3, № 1. – С. 26–38.
5. Ajiro K. An endogenous calcium-dependent, caspase-independent intranuclear degradation pathway in thymocyte nuclei: antagonism by physiological concentrations of K(+) ions / K. Ajiro, C. D. Bortner, J. Westmoreland // Experimental cell research. – 2008. – N 314 (6). – P. 1237–1249.
6. Cartmell J. W. Feed at the syndrome of chronic fatigue / John W. Cartmell // Frontier Perspectives. – 2000. – Vol. 9, N. 1. – P. 234–239.
7. Ивашкин В. Т. Оксид азота в регуляции функциональной активности физиологических систем / В. Т. Ивашкин, О. М. Драпкина // Российский журнал гастроэнтерологии, гепатологии, колопроктологии. – 2000. – Т. Х, № 4. – С. 16-21.
8. Карбашевська Н. Я. Окиснювально-антиоксидантний статус щурів за умов гіпергравітації / Н. Я. Карбашевська, С. А. Олійник, Ю. М. Білокінь та інш. // Фізіол. журнал. – 2001. – Т. 47, № 5. – С. 77–81.
9. Khan S. A. The role of nitric oxide in the physiological regulation of Ca2+ cycling / S. A. Khan, J. M. Hare // Curr. Opin. Drug Discov. Devel. – 2003. – Vol. 6, № 5. – P. 658 – 666.
10. Пат. 35792 Україна, МПК А 61В 5/145. Спосіб корекції несприятливої дії гравітаційних перевантажень в експерименті / Пикалюк В. С., Кутя С. А., Мороз Г. О., Коняєва О. І., винахідники і власники В. С. Пикалюк, С. А. Кутя, Г. О. Мороз, О. І. Коняєва. – № 200803985; заявл. 31.03.2008, опубл. 10.10.2008, Бюл. № 19, 2008.

3.3
Ваша оценка: Нет Средняя: 3.3 (10 голосов)

Замечательная статья о

Замечательная статья о медицине будущего. Желаю вам и дальше покорять космос науки, не испытывая при этом пагубного воздействия гипергравитационных перегрузок!

Глубокоуважаемая Елена

Глубокоуважаемая Елена Николаевна! Спасибо за положительный отзыв о нашей работе. В своем исследовании мы использовали крыс по двум причинам. Во-первых, лабораторные крысы это универсальный объект для моделирования в экспериментальной медицине, во-вторых, конструкционные особенности имеющейся в нашем распоряжении центрифуги не позволяют использовать более крупных животных. Что касается возраста животных, то данная работа является лишь небольшим фрагментом НИР кафедры нормальной анатомии КГМУ и в наших других публикациях отражены результаты исследования влияния гравитационных перегрузок на организм шести- и двенадцатимесячных крыс. С уважением Сергей Анатольевич Кутя.

SOME TISSUE METABOLISM MARKERS

Тема актуальна, спасибо
Лукъяненко Татьяна

Very interesting article. Why

Very interesting article. Why did your use rats and age of this animals????

Актуальность

Предложен оригинальный подход к решению антигравитационнай защиты
Партнеры
 
 
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
image
Would you like to know all the news about GISAP project and be up to date of all news from GISAP? Register for free news right now and you will be receiving them on your e-mail right away as soon as they are published on GISAP portal.