Purpose of the study. An analysis of indices of free radical oxidation and respiration of mitochondria of heart cells in a malignant process in presence of diabetes mellitus and chronic neurogenic pain in experimental animals.Materials and methods. The study included outbred female rats (n=32) and С57ВL/6 female mice (n=84). Experimental groups of rats were: intact group 1 (n=8), control group 1 (n=8) with diabetes mellitus (DM), comparison group 1 (n=8) with standard subcutaneous transplantation of Guerin’s carcinoma, main group 1 (n=8) with Guerin’s carcinoma transplanted after 1 week of persistent hyperglycemia. Experimental groups of mice were: intact group 2 (n=21), control group 2 (n=21) with a model of chronic neurogenic pain (CNP), comparison group 2 (n=21) with standard subcutaneous transplantation of melanoma (B16/F10), main group 2 (n=21) (CNP+B16/F10) with melanoma transplanted 3 weeks after the CNP model creation. Heart mitochondria were isolated by differential centrifugation. Levels of cytochrome C (ng/mg of protein), 8-hydroxy-2'-deoxyguanosine (8-OHdG) (ng/mg of protein), and malondialdehyde (MDA) (μmol/g of protein) were measured in mitochondrial samples by ELISA. Statistical analysis was performed using the Statistica 10.0 program.Results. DM in rats upregulated 8-OHdG by 6.3 times and MDA by 1.9 times (р=0.0000) and downregulated cytochrome C by 1.5 times (р=0.0053) in heart cell mitochondria, compared to intact values. DM+Guerin’s carcinoma in rats increased 8-OHdG by 14.0 times and MDA by 1.7 times (р=0.0000) and decreased cytochrome C by 1.5 times (р=0.0000), compared to intact values. CNP in mice did not affect the studied parameters in mitochondria of the heart. CNP+B16/F10 in mice increased 8-OHdG by 7.1 times and MDA by 1.6 times (р=0.0000) and decreased cytochrome C by 1.6 times (р=0.0008).Conclusions. Comorbidity (diabetes mellitus, chronic neurogenic pain) together with malignant pathology aggravates mitochondrial dysfunction of heart cells with destabilization of the respiratory chain mediated by free radical oxidation processes.

Известно, что биогенные амины (БА) участвуют в злокачественном росте, их уровень изменяется в ЦНС при болевом воздействии, однако исследований о сочетанном влиянии хронической боли (ХБ) и онкопатологии на динамику БА в головном мозге не проводилось. Цель: изучить особенности баланса БА в коре головного мозга в динамике роста меланомы, воспроизведенной на фоне ХБ. Материалы и методы. Работа выполнена на 64 мышах-самках, весом 21-22 г. Животным основной группы меланому В16/F10 перевивали под кожу спины через 2 недели после перевязки седалищных нервов. Группой сравнения служили мыши с меланомой без боли. Уровни БА: адреналина, норадреналина, дофамина (ДА), серотонина (5-НТ), гистамина, а также 5-ОИУК определяли методом иммуноферментного анализа. Результаты. У мышей с ХБ уменьшается содержание большинства БА, однако уровень ДА не изменяется. Метаболизм 5-НТ происходит с участием МАО. Развитие меланомы сопровождается увеличением содержания ДА и 5-НТ, тогда как МАО - ингибируется. Направленность сдвигов БА при развитии меланомы на фоне ХБ оказалась практически такой же, как и без неё. В то же время ХБ ограничивает накопление 5-НТ в коре мозга при меланоме, что сопровождается более агрессивным её течением. Выводы. ХБ ограничивает включение стресс-лимитирующих механизмов в головном мозге при развитии меланомы у мышей, что приводит к более агрессивному течению злокачественного процесса. Biogenic amines (BA) are known to be involved in malignant growth, and their CNS levels change in pain; however, there are no studies of combined effects of chronic pain (CP) and cancer on BA dynamics in the brain. Aim: To study features of BA balance in the cerebral cortex during melanoma growth associated with CP. Material and methods. The study included 64 female mice weighing 21-22 g. In the main groups, B16/F10 melanoma was transplanted under the skin of the back two weeks following sciatic nerve ligation. Mice with melanoma without pain were used as the control. Concentrations of BA: adrenaline, noradrenaline, dopamine (DA), serotonin (5-HT), histamine and 5-HIAA were measured with ELISA. Results. Concentrations of BAs decreased in mice with CP although DA levels did not change. 5-HT metabolism involved MAO. The development of melanoma was accompanied by increases in DA and 5-HT whereas MAO was inhibited. The direction of BA changes during the development of melanoma was the same with and without CP. At the same time, CP with melanoma limited accumulation of 5-HT in the cerebral cortex, which resulted in even more aggressive course of cancer. Conclusion. CP restricted the activation of cerebral stress-limiting mechanisms during the development of melanoma in mice, which resulted in a more aggressive course of disease.

Предупреждение метастазирования злокачественных опухолей является важной, но трудно выполнимой задачей. Целью работы была разработка способа предупреждения метастатического поражения легких у крыс с использованием диэтилбензимидазолия трийодида. Эксперименты выполнены на 38 белых разнополых крысах. Сразу после введения опухолевых клеток в подключичную вену внутрижелудочно начинали вводить диэтилбензимидазолия трийодид в разовой дозе 0,4 мг/кг. Препарат применяли в течение 8 недель по схеме: 5 дней — диэтилбензимидазолия трийодид; 2 дня — перерыв. Забой выживших животных проводили через 1,5 мес после окончания введения препарата или в преагональном состоянии. Диэтилбензимидазолия трийодид уменьшал смертность крыс обоего пола (33 против 100 %, p = 0,001). У самцов препарат в 100 % случаев ограничивал развитие метастазов, у самок — в 67 %. Причиной гибели крыс на фоне приема диэтилбензимидазолия трийодида являлось нарушение работы органов, входящих в основные оси нейрогуморальной регуляции организма: гипоталамо-гипофизарно-надпочечниковую и гипоталамо-гипофизарно-тиреоидную. Эти изменения были максимально выражены у самок в виде тиреоидной гипоплазии вплоть до инволюции железы. Диэтилбензимидазолия трийодид может быть применён для предупреждения развития метастазов в легких в эксперименте. Необходимо более глубокое изучение процессов, приводящих к летальному исходу животных на фоне приема препарата.

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.