Aim. Our aim has been to develop an experimental model of the tumor growth against the background of hypothyroidism in rats of both genders in order to study possible influence made by hypothyroidism on progression of malignant tumors of various histological types. Materials and methods. In our studies we have used 100 outbred albino rats of both genders, with an individual body mass of 150-180 g. The female rats (n=30) and the male rats (n=30) have received Mercazolil at a day dosage of 2,5 mg/100g of the body weight for 30 days. After hypothyroidism in the treated rodents had been confirmed, one group of them (15 females and 15 males) were subcutaneously inoculated with the Guerin’s carcinoma cells, and another group (covering other 15 females and other 15 males) has been undergone to transplantation of the Sarcoma 45 cells. The reference group has included the rats of both genders with subcutaneously inoculated the Guerin’s carcinoma cells (n=10 females and n=10 males) and Sarcoma 45 cells (n=10 females and n=10 males), but without reproduction of the hypothyroidism model. Upon expiration of one month, within the 3 day period, we have estimated with a radioisotope analysis (RIA) standard assay kits (Immunotech, Czekh Republic) the levels of the thyroid hormones in blood of the tested animals as follows: Triiodothyronine (T3) (pM/L), total Thyroxine (T4) (pM/L) and Thyroid-Stimulating Hormon (TSH) (μU/mL). The obtained data have been processed with Statistica 10.0. Results. Upon the treatment with Mercazolil, we have found in the females a decrease by a factor of 7,3 in the total level of Thyroxine and an increase by a factor 1,6 in the TSH level (p<0,05), while in the males we have recorded a reduction by a factor of 2 in the total level of Thyroxine and an increase by a factor of 1,5 in the TSH level (p<0,05). In this case, the average sizes of the tumors in the female rats with Guerin’s carcinoma and hypothyroidism have been found smaller than those found in the reference group as given below: upon expiration of 4 days they are 1,3 times smaller (p<0,05), upon expiration of 7 and 10 days the volumes have been found 1,4 times smaller (p<0,05); upon expiration of 14 days the volumes have been recorded to be 1,5 times less (p<0,05); upon expiration of 18 days they have been reported to be 1,3 times less (p<0,05), and upon expiration of 21 days they have been estimated to be 1,4 times less (p<0,05). As to the males with Guerin’s carcinoma and hypothyroidism, the average sizes of their tumors as against the reference group data have been recorded to be smaller as follows: upon expiration of 4 days they are found 13,3 times less; upon expiration of 7 days they have been recorded to be 7,5 times smaller; upon expiration of 10 days the volumes have been estimated to be 1,9 times less (p<0,05), and upon expiration of 14 days they have been found to be 2,6 times less. The survival rate in the female rats in the main test has been recorded to be 1,6 times higher (p<0,05) against the data in the reference group, while the survival rate in the males has not shown any significant differences therein. In the female rates with S 45 growing against the background of hypothyroidism the average sizes of the tumors have been found to be less than those identified in the reference group as follows: after 4 days, the sizes have been recorded to be 1,4 times less (p<0,05); after 7 and 10 days they have been recorded to be 1,6 and 3,2 times smaller, respectively (p<0,05); after 14 days they have been found to be 3,9 times less, and after 18 days they have been recorded to be 4,8 times less. In the males at tumor growth week stage 1, the tumor sizes have increased 3,1 times as against 4 days of the tumor growth; upon expiration of 10 days the sizes have been found to be 7,1 times greater as compared with the previous period; upon expiration of 2 weeks they have increased 1,5 times (p<0,05); upon expiration of 18 and 21 days the tumor sizes have been recorded to be greater by a factor of 2,3 and by a factor of 1,6, respectively (p<0,05). The life spans in the female rodents in the main test group has been reported to be longer by a factor of 1,8 (p<0,05) than it has been the case with the reference group, and the average life span in the males has reached 21 days. Conclusion. We have revealed that in the female rates diagnosed with hypothyroidism the sizes of the subcutaneous tumor nodes of Guerin’s carcinoma and S 45 show slower progression as against that recorded in the reference group, and the life span recorded in the above rodents has been found as significantly longer, while in the male rats with hypothyroidism we have observed an irregular, slower, progression of the tumor nodes of Guerin’s carcinoma and S 45 within the period of 14 days, but subsequently we have detected the same progression rate as it is the case with the reference group data.

Введение. Печень по количеству, плотности митохондрий один из самых богатых органов, который также является критическим местом для множества метаболических путей. Цель исследования - изучение показателей апоптоза в митохондриях печени самок мышей линии С57ВL/6 при самостоятельном росте меланомы В16/F10 и на фоне коморбидной патологии - хронической нейрогенной боли. Методика. В эксперименте использовали мышей-самок (n=168) линии С57ВL/6. Группы: интактная (n=21); контрольная (n=21) - создание модели хронической нейрогенной боли (ХНБ), путем двусторонней перевязки седалищных нервов; группа сравнения (n=63) - подкожная трансплантация меланомы B16/F10; основная группа (ХНБ+B16/F10) (n=63) - подкожная трансплантация меланомы В16/F10 через 3 нед после моделировия ХНБ. В митохондриях печени методом ИФА определяли концентрацию: цитохрома С (нг/г белка), каспазы 9 (нг/г белка), Bcl-2 (нг/г белка), AIF (нг/г белка), кальция (Са 2+) (мМоль/г белка). Результаты. В митохондриях клеток печени через 1 нед роста меланомы относительно интактных значений фиксировали нарастание уровней AIF в 2,2 раза, цитохрома С в 1,7 раза (р<0,05) и снижение каспазы 9 в 2,0 раза; через 3 нед - падение кальция в 4,7 раза, AIF в 7,1 раза и цитохрома С в 1,7 раза (р<0,05) и накопление каспазы 9 - 1,6 раза (р<0,05). Развитие опухоли при ХНБ через 1 нед сопровождалось уменьшением концентрации AIF в 29,3 раза и цитохрома С в 2,0 раза по сравнению с контрольными значениями (ХНБ). Через 3 недели роста меланомы на фоне ХНБ фиксировали снижение уровней AIF в 6,6 раза, цитохрома С в 4,7 раза и кальция в 32,8 раза, уровень каспазы 9, напротив, повышался в 1,5 раза (р<0,05). Заключение. Наличие коморбидной патологии - ХНБ при опухолевом процессе способствует раннему возникновению нарушений в электронно-транспортной цепи митохондрий клеток печени. Background. The liver is one of the richest organs in terms of the number and density of mitochondria; it is also a critical site for many metabolic pathways. The aim of the study was to analyze indicators of apoptosis in liver mitochondria in female С57ВL/6 mice with B16/F10 melanoma growing alone and in presence of chronic neurogenic pain. Methods. Female С57ВL/6 mice (n=168) were studied. Animals were divided into groups: intact group (n=21); controls (n=21) with a model of chronic neurogenic pain (CNP) created by bilateral sciatic nerve ligation; comparison group (n=63) with subcutaneous transplantation of B16/F10 melanoma; main group (CNP+B16/F10) (n=63) with subcutaneous transplantation of B16/F10 melanoma 3 wks after modeling CNP. Cytochrome C (ng/g protein), caspase-9 (ng/g protein), Bcl-2 (ng/g protein), AIF (ng/g protein), and calcium (Ca2+) (mmol/g protein) were measured by ELISA in the liver mitochondrial fraction. Results. After 1 wk of melanoma growth, AIF increased by 2.2 times, cytochrome C increased by 1.7 times (p<0.05), and caspase-9 decreased by 2.0 times compared to the intact group values. After 3 wks, calcium decreased by 4.7 times, AIF by 7.1 times, cytochrome C by 1.7 times (p<0.05), and caspase-9 increased by 1.6 times (p<0.05). After 1 wk, tumor development in the presence of CNP was accompanied by decreases in AIF by 29.3 times and cytochrome C by 2.0 times, compared to control CNP values. After 3 wks of melanoma growth in presence of CNP, AIF decreased by 6.6 times, cytochrome C by 4.7 times, and calcium by 32.8 times. Caspase-9, on the contrary, increased by 1.5 times (p<0.05). Conclusions. The presence of CNP comorbidity during the tumor development facilitates earlier occurrence of disorders in the electron transport chain of hepatocyte mitochondria.

The aim is to evaluate the physiological parameters of the efficacy of cardiac mitochondria transplantation in male mice with chronic neurogenic pain and B16/F10 melanoma growth. Materials and methods. Male mice (n=37) of the C57BL/6 line were used in the research work. The animals covered experimental groups as follows: mice with chronic neurogenic pain (CNP) + B16/F10 melanoma (n=27); mice with CNB + B16/ F10 melanoma + mitochondrial therapy (MC therapy) (n=10). Mitochondria were isolated from the heart of an intact rat with the use of differential centrifugation. An introduction of mitochondria to mice was carried out daily intraperitoneally at a dose of 3.3 mg of protein for 3 weeks. Statistical analysis of the results is carried out with the Statistica 10.0 software. Results. On day 21 (week 3) of the experiment, macroscopically in the melanoma tissue in the group of animals with MC therapy, there were 2.5 times more necrosis cases than in the group without MC therapy. During the examination of the internal organs, no metastases were detected in the animals treated with MC therapy, while in 100% of the animals without MC therapy metastases were found in the lungs and in 95% of them in the spleen. In the animals received MC therapy, there was no damage to the heart muscle in 75% of the cases, while in the group of the animals without MC therapy, the presence of lesions in the form of bruises on the surface of the heart was macroscopically detected in 100% of the animals. Conclusion. Thus, intraperitoneal transplantation of intact heart mitochondria contributed to the prevention of myocardial infarction and metastases to internal organs in the C57BL/6 male mice with B16/F10 melanoma growing against the background of chronic neurogenic pain.

The aim is to study some mechanisms of regulation of apoptosis and self-organization in the mitochondria in the heart cells in female mice during the growth of experimental melanoma B16/ F10 linked with chronic neurogenic pain as comorbid pathology.

Topicality. An increase in the incidence of malignant tumors progressing against the background of various comorbid pathologies determines the need to study the mutual influence of pathological processes using experimental modeling. Such models, for example, can reproduce the tumor growth modified by the comorbid condition of diabetes mellitus, the incidence rate of which is now increasing exponentially. In this case, a clear demonstration of changes in the structure of organs outside the zone of the primary tumor node, using data on the direct growth of the tumor and the content of diabetes markers therein and in the perifocal zone, can serve as an evidence-based argument for the implementation of the mechanism of modified tumor progression. The aim of our research work is to assess the state of the histological structure of some internal organs (the kidneys, the ovaries, the peritoneum) when modeling the main pathological process in animals, the growth of Guerin’s carcinoma, against the background of a comorbid state of experimental diabetes. Materials and methods. We used 32 outbred male and female rats weighing 180-220 g to reproduce the model of experimental diabetes by a single intraperitoneal injection of alloxan at a dosage of 150 mg/kg of body weight. One week after the production of persistent hyperglycemia in the range of 25.4±1.2 mmol/l, the animals were transplanted with Guerin’s carcinoma subcutaneously in the region of the right shoulder blade. Upon expiration of 2 weeks, the animals were decapitated, and the harvested organs were prepared according to the practice stages of morphological preparation for staining sections with hematoxylin-eosin, followed by morphological examination of the structure with the use of the Leica DM LS2 microscope with the Olympus optical. C-5050 Zoom video camera and the Morfotest software. Photographing was carried out with magnification x10, x40, x100. Results. Our study of the morphological portrayal of the ovary, the kidney, the visceral and parietal peritoneum bears witness to the identity of the changes, consisting in a total metastatic lesion and abnormal transformation of the normal structure of all the studied organs only in the female rats, when modeling the comorbid state of diabetes mellitus. At the same time, the aggressive nature of the tumor progression was manifested in the blood filling of the vessels and hemorrhage, followed by the release of tumor cells, their settlement, the enhanced proliferation, the formation of strands and compaction of cell aggregations throughout the volume of the organ. Some gender specific features of the tumor progression were noted, which were found in the female rats along the path of active metastasizing in case of small primary tumors, and in the male rats along the path of stimulating the growth of the primary focus without metastasizing. It was revealed that these differences are associated with different degrees of saturation of the tumor and perifocal zone with glucose, and they are determined by the state of the insulin/insulin-like growth factor (IGF) axis. Conclusion. Morphological examination of the organs affected by metastatic Guerin’s carcinoma in the female rats with diabetes mellitus makes it possible to detect not only the synergy of both pathological processes, but also a powerful pro-oncogenic effect of the comorbid state of diabetes in the implementation of the tumor growth program.

studying AIF levels in mitochondria in cells of the heart and various organs in female mice with the growth of experimental melanoma B16 / F10 and comorbid pathology

Mitochondria are dynamic organelles which constantly change their shape, size, and location within the cells. Mitochondrial dynamics is associated with mesenchymal metabolism or epithelial-mesenchymal transition to regulate the stem cell differentiation, proliferation, migration, and apoptosis. The transfer of mitochondria from one cell to another is necessary to improve and maintain homeostasis in an organism. Mitochondrial transplantation is a therapeutic approach that involves an introduction of healthy mitochondria into damaged organs. Recent evidence data have shown that the physiological properties of healthy mitochondria provide their ability to replace damaged mitochondria, with suggesting that replacing damaged mitochondria with healthy mitochondria may protect cells from further damage. Moreover, mitochondria can also be actively released into the extracellular space and potentially be transferred between the cells in the central nervous system. This increased interest in mitochondrial therapy calls for a deeper understanding of the mechanisms, which build the basis for mitochondrial transfer, uptake, and cellular defense. In this review, questions related to the involvement of mitochondria in the pathogenesis of cancer will be discussed. Particular attention will be paid to mitochondrial transplantation as a therapeutic approach to treat the mitochondrial dysfunction under some pathological conditions.

The aim is to evaluate the pathophysiological parameters of the efficacy of liver mitochondrial transplantation in animals with B16/F10 melanoma. Materials and methods. In our experiment we used female and male mice of BALB/c Nude strain (n=28). Experimental groups were as follows: the reference group (n=14) with B16/ F10 melanoma; the main group (n=14) with B16/F10 melanoma + mitochondrial therapy (MC therapy). Statistical analysis of results was carried out with the Statistica 10.0 software. Results. The subcutaneous tumor in the mice of both sexes became detectable on day 5 from the time of the tumor inoculation, and the regressive effect produced by MC therapy was recorded in the males beginning with day 8 of the tumor growth. At the end of the experiment, on day 22, the difference in the average volumes of the tumor node was reported to be 3.2 times, i.e. a significant inhibition of the tumor growth in the group of the males with MC therapy was revealed. In the females on day 5 of the tumor growth, differences in the volume of the tumor focus between the reference group and the group with MC therapy were not recorded, however, a statistically significant difference was found in the sex-related comparison of the groups of the animals with MC therapy. It was determined that in the females with MC therapy, the area of the tumor spot during that period (5 days) was 1.4 times (p<0.05) less than that in the corresponding group of the males. On day 8, in the females completed MC therapy, the tumor has not yet concentrated into a solid structure, but remained as a flat tumor entity, and only by day 12 the tumor has formed from a flat structure into a volumetric tumor type. As a result, by the end of the experiment, on day 22, smaller volumes of the tumor nodes remained in the group of females treated with MC therapy, and the difference with the reference group was 2.7 times (p<0.05). Conclusion. Thus, within the framework of the experiment, it has been found that the application of mitochondrial therapy using allogeneic liver mitochondria in the BALB/c Nude mice with B16/F10 melanoma retards the tumor growth in the mice of both sexes.