CC BY
20УДК 616 - 092.9:615.383:616.831-036.88
THE INFLUENCE OF "MEDIATORY SUBSTANCES" OF FETAL CELLS ON CHANGES IN THE TRANSAMINASE ENZYME ACTIVITY IN BLOOD SERUM OF RATS AFTER FATAL HYPOBARIC HYPOXIA
YERMENTAEVA LAZZATNURMAKHANBETOVNA
candidate of medical sciences, senior lecturer of the Pathological Physiology Department of V.G.Korpachev, JSC "Astana Medical
University", Astana, Kazakhstan. E-mail: ermentaeva 70@mail. ru
AITBAYEVA ZHAINA BAIDULLAYEVNA
doctor of medical sciences, professor of the Pathological Physiology
Department of V.G.Korpachev, JSC "Astana Medical University", Astana, Kazakhstan.
AKPOLATOVA GULNUR MOMYNOVNA
candidate of medical sciences, docent of the Pathological Physiology Department of V.G.Korpachev, JSC "Astana Medical
University", Astana, Kazakhstan.
BEGLAROVA GULSHAHAR ERKINOVNA
candidate of medical sciences, professor of the JSC "Astana
Medical University", Astana, Kazakhstan.
ABSTRACT
The experiments were carried out on 86 white outbred rats, which had single and double transplantation of"mediatory substances" of fetal cells in the postresuscitation period. The transaminase enzyme activity of blood serum in dynamics of postresuscitation period was studied in the experiments. The results of our study suggest that the application of "mediatory substances" of fetal cells does not aggravate the severity of postresuscitation period after 4 minutes of clinical death caused by hypobaric hypoxia. In addition, calculated De Ritis coefficients in the experimental groups do not exclude the probability of activation of adaptive mechanisms under the influence of the factors, causing the development of extreme and terminal conditions.
Keywords: "mediatory substances" of fetal cells, postresuscitation period, transaminase enzymes, De Ritis coefficient, cell therapy, adaptation, hypobaric hypoxia.
ВЛИЯНИЕ «МЕДИАТОРНЫХ ВЕЩЕСТВ» ФЕТАЛЬНОЙ КЛЕТКИ НА ИЗМЕНЕНИЕ АКТИВНОСТИ ТРАНСАМИНАЗНЫХ ФЕРМЕНТОВ СЫВОРОТКИ КРОВИ У КРЫС ПОСЛЕ СМЕРТЕЛЬНОЙ
ГИПОБАРИЧЕСКОЙ ГИПОКСИИ
ЕРМЕНТЕВА ЛАЗЗАТ НУРМАХАНБЕТОВНА
кандидат медицинских наук, старший преподаватель кафедры патологической физиологии им.В.Г.Корпачева.
АО «Медицинский университет Астана», г.Астана
(Казахстан) E-mail: ermentaeva 70@mail. ru
АЙТБАЕВА ЖАЙНА БАЙДУЛЛАЕВНА
доктор медицинских наук, профессор кафедры патологической физиологии им.В.Г.Корпачева.
АО «Медицинский университет Астана», г.Астана
(Казахстан)
АКПОЛАТОВА ГУЛЬНУР МОМЫНОВНА
кандидат медицинских наук, доцент кафедры патологической
физиологии им.В.Г.Корпачева.
АО «Медицинский университет Астана», г.Астана
(Казахстан)
БЕГЛАРОВА ГУЛЬШАХАР ЕРКИНОВНА
кандидат мед.наук, ассоциированный профессор, АО «(Медицинский Университет Астаны» г.Астана (Казахстан)
АННОТАЦИЯ
Эксперименты были выполнены на 86 белых беспородных крысах, которым в постреанимационном периоде осуществляли однократную и двукратную трансплантацию «медиаторных веществ» фетальных клеток. В опытах изучали активность трансаминазных ферментов сыворотки крови в динамике постреанимационного периода. Результаты наших исследований свидетельствуют о том, что использование «медиаторных веществ» фетальных клеток не усугубляет тяжести течения постреанимационного периода после 4-минутной клинической смерти, вызванной гипобарической гипоксии. Кроме того, рассчитанные в опытных группах коэффициенты Де Ритиса
не исключают вероятность активации адаптивных механизмов при действии факторов, вызывающих развитие экстремальных и терминальных состояний.
Ключевые слова: «медиаторные вещества» фетальной клетки, постреанимационный период, трансаминазные ферменты, клеточная терапия, гипобарическая гипоксия.
Relevance. In recent years, researches directed on studying the possible ways of usage cellular therapy become widespread [9, 6, 1, 11, 12]. One of them is the application of so-called "mediatory substances" from fetal cells, which are obtained by dissociation of cells.
During the process of getting the fetal cells, appearsupernatant, made from protein-peptide complex of active substances, such as interleukins 2, 6, 10, vascular -endothelial factor, peptide hormones [4, 8].
By several studies, it has proventhat their usage could help for rehabilitation of metabolism of organism and normalization of disordered haemostasis in irregularly pathology. Moreover, the "mediatory substances" of fetal cells are powerful regulators that affect body cells of recipient, which remedy their function and interaction, at the same time possess a directional gentle influence on the regulatory fetal functions, which facilitates to the restoration of body's normal functioning [5]. A number of authors have obtained results indicating an improvement of animals' conditionwith toxic hepatitis, stress caused by hypoxia and immobilization, diabetesmellitus, hepatic failure [10, 2, 3, 8].
However, it must be admitted that the method of therapy using "mediatory substances" of fetal cells, has not receive detailed researches and pathogenic substantiation yet. Experimental data, studying the mechanisms ofactions in "mediatory substances"of fetal cells is obviously
insufficient, and it requires further studies. Because of that, it was interesting to us to study experimentally influence of "mediatory substances" of fetal cells on changes in the transaminase enzyme activity, getting from blood serum of rats after fatal hypobaric hypoxia.
Purpose of the study. The aim of the research was to study the transaminase enzyme activity from blood serum in dynamics of postresuscitation period in conditions of single and double injection of "mediatory substances", obtained in the process of isolating fetal liver cells.
Materials and methods. The experiments were carried out on 86 adult white rats. Experimental animals were divided into 4 observation groups: I group - intact animals (n = 33); Ilgroup - control animals (n = 23); III group -experienced with a single injection of "mediatory substances" of fetal cells in the recovery period after resuscitation (n = 22). IV group -resuscitated animals with the double injection of "mediatory substances" of fetal cells (n = 8).
The increasing to 11,500 m was simulated on experimental animals within 3 minutes with the Komovski apparatus with the speed of 107 m / sec [7]. By the way, during the process was recorded the time, when starts first convulsions and agony breathing, which come as a result of the barometric pressure decrease. The last recorded breath action correspondedas the beginning of clinical death, which shows 4 minute duration in our experiment. Resuscitation consisted ofthe external cardiac massage and operated hardware ventilation of the lungs using the "Breathe RP-3" device. In this series of experiments, the injection of "mediatory substances" of fetal liver corresponded to 30 minute from the beginning of the implementation of resuscitation measures and 7 days of post resuscitation period. The substrate was obtained, during the process of getting the fetal cells, as supernatant, made from protein-peptide complex of active substances
(mediatory substance). The dose of injected ""mediatory substances" was 0.3 mL / kg [8]. Searching of the transaminase enzymes (ALT, AST) was carried out after 14 days of resuscitation on standardized biochemical analyzers. The research results were processed statistically using the Student's t-criterion. Digital processing of the material was carried out on a computer using a set of Microsoft Excel's standard programs. The main results of the research are presented in Table 1.
The research material determined that after14 days, successfully, was completed cardiopulmonary resuscitation in the control group (II) the indicators of transaminase activity (AST and ALT) in animals' serum had no significant differences with the intact group.
In the III experimental group of animals, in which at the 30 minutes of post resuscitation period was performed single intraperitoneal injection of "mediatory substances" of fetal cells, found that after 14 days the level of ALT'sactivity was equal to 0,69 □ 0,05 mmol / l x h. Where in the studied indicators' values were significantly lower than in control (p1 <0.05). At the same time on the background of the single application of "mediatory substances" of fetal cells, the AST's activity was increased to 1,50 ± 0.05 mmol / l x h.
Under the double application of "mediatory substances" of fetal cells (IV group), the data showing a decrease in the ALT's activity compared with the control of 2.2 times were obtained.
Table 1 - Changes in the transaminase enzyme activity in animals' blood serum in dynamics of postresuscitation period on the background of single and double application of fetal-cell therapy.
Group number / Number of studies
Studied
Indicators Res Ilgroup Illgroup IV group
(I group) earch days
ALT 14 0,86+0,04 0,69+0,05 0,39+0,06
(norm days p<0,05 pi<0,05 P1<0,001
0,96D0,04 P12<0,05
mmol / l h)
AST 14 1,43+0,06 1,50+0,05 1,20+0,03
(norm days p>0,05 pi>0,05 P1<0,02
1,36D0,05 P2<0,001
mmol / l h)
Notes:
1. reliabilityof differences compared with Igroup;
2.p1 significance of differences compared with the control;
3.p2 significance of differences compared with IVgroup.
The indicators of enzymatic activity of ALTwere also lower than the values registered in the III group of observation, wherein the "mediatory substances" therapy for animals was carried out only once (0.39 ± 0.06 mmol / l x h vs. 0.69 ± 0.05 mmol / l x h p2 <0.001).
During this period in the IVgroup of the research found that the AST's activity was lower than in controlled animals with a single injection of the "mediatory substances" of fetal cells (III group) and amounted to 1,20D0,03 mmol / l x h (p1 □ 0.02).
Analysis of the results showed that in the dynamics of recovery period there was a change in the ratio of transaminase enzymes in blood serum. Thus, during the research period, the De Ritis coefficient in animals by using "mediator substances" of fetal liver tissue (III and IV groups) exceeded the values, obtained in the comparison groups (I-intact and II-control).
Conclusions. To conclude results of our research, we relied on the data stated in the research reports of I.M.Roslyi and co-authors (2002,
2003), in which were discussed the possible causes, mechanisms and ways
86
of appearance of blood enzymes. Particularly, the authors point out that a high level of enzymes identified in animals is not always evidence of apparent cytolysis. The ratios of transaminase enzymes in blood are more significant, which also may reflect the metabolic changes of adaptive nature. The results of our research suggest that the usage of "mediatory substances" of fetal cells do not aggravate the severity of post resuscitation period after 4 minutes of clinical death caused by hypobaric hypoxia. In addition, calculated De Ritis coefficients in the experimental groups do not exclude the probability of activation of adaptive mechanisms under the influence of the factors, causing the development of extreme and terminal conditions. It can be assumed that a body is additionally brought in biologically active materials (hormones, alpha-fetoprotein, growth factors, peptides, cytokines, immunoglobulins, electrolytes and others), which stimulate body's sanogenetic mechanisms.
LITERATURE
13. Айтбаева Ж.Б.,Тимурин Д.Н., Торопеев А.В. Влияние фетальных нейроцитов на динамику показателя общего состояния крыс-самцов //Материалы V международной дистанционной научно-практической конференции «Новые технологии в медицине-2008» // Bulletin. - 2008. - Vol.3 - №1. - С. 9 - 10.
14. Акполатова Г.М. Влияние «медиаторных веществ» фетальных клеток на изменение адаптивных реакций организма при стрессе, вызванном иммобилизацией и гипоксией (экспериментальное исследование): автореф. ... канд. мед. наук: 14.00.16 - патологическая физиология. - Астана: КазГМА, 2008. - 22 с.
15. Букеева Ж.К. Влияние «медиаторных веществ» фетальных клеток на течение острого и хронического гепатита у крыс
(экспериментальное исследование):автореф... канд. мед. наук: 14.00.16 - патологическая физиология. -Астана: КазГМА, 2007. - 19 с
16. Доскалиев Ж.А., Мустафин А.Х., Жетимкаринова А.Д., Стикеева Р.К. и др. Состояние симпатико-адреналовой системы после обширных резекций печени с трансплантацией медиаторов фетоткани // Астана медициналык журналы. - 2006. - №2. - С. 154-155.
17. Доскалиев Ж.А., Хамзина Н.К., Стикеева Р.К. и др. Механизм действия «медиаторов» фетальных клеток //Валеология.-2008.-№1. -С. 53-55.
18. Мухамеджанова К.М. Влияние клеточной терапии на процессы заживления ожоговой раны в постреанимационном периоде (экспериментальное исследование):автореф. ... канд. мед. наук: 14.00.16 - патологическая физиология.- Астана, 2007. - 22 с.
19. Рахимбердиев Д.С. Влияние суттигена на течение постреанимационной энцефалопатии: автореф. ... канд. мед.наук:
20.12.04.- Астана: КазГМА, 2004.-17 с.
20. Стикеева Р.К. Влияние «медиаторов» фетальных клеток на состояние липидного обмена при печеночной недостаточности // Астана медицинальщ журналы. - 2007. - №9 (45). - С. 179-181.
21. Тажибаева Д.С. Патогенетическое обоснование клеточной терапии для профилактики и лечения постреанимационной болезни и постгипоксической энцефалопатии: автореф. ...докт. мед.наук:
17.06.05.- Астана: КазГМА, 2005.-С.50
22. Таржанова Д.Ш.Влияние «медиаторов» фетальной ткани печени на течение экспериментального сахарного диабета// (экспериментальное исследование): автореф. ... канд. мед.наук: 14.00.16 - патологическая физиология. -Астана: АО «АМУ», 2009. - 21 с.
23. Dani A., Bartoli G.M., Galeotti T. The operation of the malate-aspartate shuttle in the reoxidation of glycolytic NADH in slices of fetal rat liver // BiochimBiophysActa. 1977 Dec 23;462(3):781
24. Hu X., Yang T., Li C., Zhang L., Li M., Huang W., Zhou P. Human fetal hepatocyte line, L-02, exhibits good liver function in vitro and in an acute liver failure model// Transplant Proc. 2013 Mar;45(2):695-700. doi: 10.1016/j.transproceed.2012.09.121.
