Исследование проводили на мышах линии C57/Bl6 обоего пола, которым по стандартной методике перевивали меланому В16/F10. В динамике роста опухоли, через 1,2,3 и 4 недели в гомогенатах гипоталамуса, гипофиза, гонад, надпочечников и щитовидной железы определяли регуляторные и периферические гормоны. Рост перевивной меланомы В16/F10 у мышей обоего пола вызывает изменение функционирование основных регуляторных осей организма - гипоталамо-гипофизарно-надпочечниковой (ГГН), тиреоидной (ГГТ) и гонадной (ГГГ), оказывая влияние как на центральные, так и на периферические звенья регуляторных осей. Реализация адаптационных реакций на рост опухоли имеет особенности, как в сроках наступления, так и в интенсивности проявления, зависящие от половой принадлежности животных. Ключевые слова: Меланома В16/F10, регуляторные оси организма, гипоталамо-гипофизарно-надпочениковая, гипоталамо-гипофизарно-тиреоидная, гипоталамо-гипофизарно-гонадная ось, мыши C57Bl/6.

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.

Introduction . Cytochrome C in mitochondria transfers electrons from complex III to complex IV, and it is a signaling molecule in the apoptosis realization. The objective was to evaluate the level of cytochrome C in cell mitochondria in various organs of female mice with standard and stimulated growth of experimental B16/F10 melanoma. Methods and materials . The experiment was performed on female C57BL/6 mice (n=168). The groups were: intact animals (n=21); controls with a model of chronic neurogenic pain (CNP) (n = 21); group M — standard B16/F10 melanoma transplantation (n=63), group CNP + M — B16/F10 melanoma transplantation 3 weeks after CNP model creation (n=63). The level of cytochrome C (ng / mg protein) were measured by ELISA (Bioscience, Austria). Statistical analysis of results was performed using the «Statistica 10.0» program. Results . After 1 week of standard melanoma growth, an increase in the level of cytochrome C by 2.7 and 1.7 times was detected in mitochondria of the brain and liver; by the 3rd week, it decreased in the liver and skin by 1.7 times. In melanoma mitochondria, the level of cytochrome C was lower than in the skin of intact animals: by 2.5 times after week 1, by 4.5 times after week 2, and by 4.6 times after week 3. After 1 week of stimulated melanoma growth, the level of cytochrome C decreased compared control values: by 2.2 times in the brain, by 1.9 times in the liver, by 1.4 times in the skin; by week 3, it decreased by 4.8 times in mitochondria of the brain, by 4.7 times — in the liver, by 2.3 times — in the heart, by 1.9 times — in the skin. In melanoma mitochondria, the level of cytochrome C was lower than in the skin of intact animals: by 15.3 times after week 1, by 10.3 times after week 2, and by 8.8 times after week 3. Conclusion . Low level of cytochrome C were found in melanoma mitochondria in standard and stimulated tumor growth. The data can be used in the experiment and in clinic for using exogenous cytochrome C as an agent slowing down the malignant process.

Introduction. The influence of chronic neurogenic pain on local levels of growth factors in the lung and the risk of lung cancer development has been little studied. The purpose of the study was to analyze the levels of VEGF, TGF-β, IGF-I, IGF-II, FGF-21 and receptors of VEGFR2, TGF-βR2 in the lungs of white outbred rats with chronic neurogenic pain after intravenous injection of M1 sarcoma. Material and Methods . A total of 28 white outbred male rats weighing 200–250 g were divided into 4 groups: 1 – sham-operated animals (control group) (n=7); 2 – animals with chronic neurogenic pain (n=7); 3 – sham-operated animals with intravenous injection of M1 sarcoma (n=7); 4 – rats with intravenous injection of M1 sarcoma in presence of chronic neurogenic pain (n=7). Animals were decapitated, the lungs were harvested on ice; 10 % cytosolic fractions were prepared in 0.1 M potassium phosphate buffer pH 7.4 containing 0.1 % Tween-20 and 1 % BSA. Levels of VEGFR2, TGF-β and receptors of TGF-βR2, IGF-I, IGF-II (CUSABIO BIOTECH Co., Ltd., China) and FGF21 (BioVender, Czech Republic) were determined by ELISA. The data were statistically processed using Statistica 10.0 software. Results . Multifocal tumors of sarcoma in the lungs were determined only in the group of rats with chronic neurogenic pain after intravenous transplantation of M1 sarcoma. In the lungs of sham-operated animals, tumor foci were not detected after M1 sarcoma transplantation. Lung tissues with M1 growth and presence of chronic neurogenic pain demonstrated decreased VEGF-А levels with increased concentrations of TGFβ, IGF-I, IGF-II and FGF-21. Chronic neurogenic pain directly or indirectly influenced levels of some growth factors in the lung, and altered cell homeostasis making possible transplantation of M1 sarcoma into the lung.

Purpose of the study . Studying the dynamics of growth factors (GF) in urokinase (uPA)-deficient mice with chronic neurogenic pain (CNP) and B16/F10 melanoma. Materials and methods . Levels of VEGFA, VEGFC, sVEGFRl, sVEGFR3, IGF1, IGF2, TGFp1 and FGF21 were determined by ELISA in tumors, perifocal tissues (PT) and the skin of male and female С 57 BL/6 mice (with a normal genome, n = 75) and C57BL/6-Plautm1.1BugThisPlauGFDhu/GFDhu mice (uPA-deficient animals, n = 46) at 3nd week of the carcinogenesis and CNP. Results . The skin of intact uPA-deficient mice demonstrated higher GF levels than in C57BL/6 mice, but VEGF- А and TGF-p1 in males (unlike females) were 4.4 and 5 times lower than in C57BL/6 males. This changes were generally similar in the skin of C57BL/6 mice with CNP. uPA-deficient females showed elevated GF in PT, especially VEGF- А and IGF1 — by 21.5 and 8.1 times, respectively in simultaneously CNP and growth of the melanoma. uPA gene-knockout males had similar changes in GF, although less marked. The levels of all studied GF in tumor tissue were lower than levels in PT in both males and females, except for VEGFA in males — 5.6 times higher in tumor tissue. Changes in PT of C57BL/6 mice were similar: maximally increased levels of all GF, especially VEGF, IGF and TGF-p1 — in females on average by 6.2, 15.9 and 5.5 times, respectively, in males by 9.4, 5.9 and 6.7 times, respectively, compared to the skin levels. While the absolute values of GF concentrations and the intensity of changes were higher than in uPA-deficient mice. Conclusion . In general, skin levels of GF in intact uPA-deficient mice were similar to the levels in mice without uPA-deficient with CNP. The GF dynamics was analogous in mice of both lines at simultaneously CNP and growth of the melanoma, but the intensity of changes in mice without uPA-deficient was significantly higher implying a synergic effect of CNP and paracrine influence of melanoma on the GF levels.

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.

Метастазирование в печень - частый признак прогрессирования злокачественного процесса, механизм которого до конца не изучен. Цель - изучение особенностей функционирования основных систем нейрогуморальной регуляции: надпочечниковой (ГГНС), тиреоидной (ГГТС) и гонадной (ГГГС), на этапах метастазирования саркомы 45 (С45) в печень. Методика. Работа выполнена на 43 белых крысах-самцах. Через 1, 2, 5, 7 нед. от момента введения клеток саркомы 45 в дислоцированную под кожу селезенку. Органы взвешивались, в сыворотке крови (СК) методом РИА исследовали уровни фолликулостимулирующего гормона, лютеинизирующего гормона (ЛГ), тиреотропного гормона, адренокортикотропного гормона, кортизола, альдостерона, общей формы тестостерона, свободной и общей форм тироксина и трийодтиронина (Тсв и Т); методом ИФА - эстрона, эстрадиола, свободного тестостерона. Результаты. Метастазирование в печень сопровождалось активацией с последующим истощением ГГНС со снижением в 1,8 раза уровней кортизола и альдостерона в крови; значительной активацией ГГГС (пятикратное увеличение ЛГ в крови и уменьшение в 1,7 раза массы семенников) вследствие гипотестостеронемии (в 9,7 раза) на фоне гиперэстрадиолемии (в 2,3 раза); активацией ГГТС с формированием «low T» синдрома (в 4,6 раза). Заключение. Процесс метастазирования в печень - системная патология, в основе которой лежат глубокие нарушения работы основных систем регуляции организма. Aim: To evaluate functioning of three major systems of neurohumoral regulation, adrenal (HPA), thyroid (HPT), and gonadal (HPG) axes, in liver metastasis based on weights of organs and blood levels of hormones. Methods: The study included 34 white male rats. Organs were weighted at 1, 2, 5, and 7 weeks after S45 sarcoma cell injections into the subcutaneously transferred spleen; blood serum levels of FSH, LH, TSH, ACTH, cortisol, aldosterone, total testosterone, free and total thyroxine, and triiodothyronine were measured by RIA; estrone, estradiol, and free testosterone concentrations were measured by ELISA. Results. The process of liver metastasis was accompanied by activation and following exhaustion of the HPA axis with 1.8 time decreases in blood levels of cortisol and aldosterone , significant activation of the HPG axis (5-fold increased LH level and 59% decreased testicular weight) due to hypotestosteronemia (9.7 times) and hyperestradiolemia (2.3 times), and activation of the HPT axis with the low T3 syndrome (4.6 times). Conclusion. Liver metastasis is a systemic pathology based on profound dysfunction of the major regulatory systems.

Метастазирование в печень - частый признак прогрессирования злокачественного процесса, механизм которого до конца не изучен. Цель – изучения особенностей функционирования основных систем нейро-гуморальной регуляции: надпочечниковой (ГГНС), тиреоидной (ГГТС) и гонадной (ГГГС), на этапах метастазирования саркомы 45 (С 45) в печень. Методика. Работа выполнена на 43 белых крысах-самцах. Через 1, 2, 5, 7 нед от момента введения клеток саркомы 45 в дислоцированную под кожу селезенку. Органы взвешивались, в сыворотке крови (СК) методом РИА исследовали уровни фолликулостимулирующего гормона, лютеинизирующего гормона (ЛГ), тиреотропного гормона, адренокортикотропного гормона, кортизола, альдостерона, общей формы тестостерона, свободной и общей форм тироксина и трийодтиронина (Т3св и Т3); методом ИФА - эстрона, эстрадиола, свободного тестостерона. Результаты. Метастазирование в печень сопровождалось активацией с последующим истощением ГГНС со снижением в 1,8 раза уровней кортизола и альдостерона в крови; значительной активацией ГГГС (пятикратное увеличение ЛГ в крови и уменьшение в 1,7 раза массы семенников) вследствие гипотестостеронемии (в 9,7 раза) на фоне гиперэстрадиолемии (в 2,3 раза); активацией ГГТС с формированием "low T3" синдрома (в 4,6 раза). Заключение. Процесс метастазирования в печень - системная патология, в основе которой лежат глубокие нарушения работы основных систем регуляции организма.

Objective : to investigate the time course of changes in the early biomarkers of acute kidney injury in patients with clinically localized cancer during partial nephrectomy, as electively indicated, under thermal ischemia with prior epidural block. Materials and methods . To analyze the nephroprotective effect of an epidural block in kidney resection with warm ischemia, markers of acute kidney injury (cystatin C, interleukin 18, NGAL, L-FABP and KIM-1) were studied by ELISA in the blood and urine of 35 patients with local cancer with an epidural block (main group) and 37 patients with local cancer without an epidural block (control group) before surgery and 40 min after its beginning and on days 1 and 3 of the postoperative period. All patients were divided into 2 groups by the levels of cystatin C in the blood serum: 1000 ng/ml and lower, and over 1000 ng/ml. Results. Epidural block during the perioperative period in kidney resection with warm ischemia for patients with local cancer had an obvious nephroprotective effect allowing maintaining the initial renal functional parameters, in contrast to the standard disease management.

Background: Skin cells are capable of the local synthesis of sex and regulatory hormones which makes it an analogue of the hypothalamic-pituitary-gonadal system and facilitates independent melanoma hormonogenesis. Aims: to study the level of hormones in the tumor, perifocal zone, the skin is unaffected, and the pituitary gland in the dynamics of growth of melanoma B16/F10 in female C57BL/6 mice. Materials and methods: Changes in the local steroidogenesis in the dynamics of the growth of transplanted В16/F10 melanoma (from week 1 to week 4) were studied in 40 female С57BL/6 mice. Hormones were determined by standard ELISA methods and radioimmunoassay in homogenates of tumor, surrounding tissues and intact tissues. Results: Tumor and then surrounding tissues showed an increase in estrogens, mostly due to estrone increase, with the development of relative androgen deficiency primarily in tumor and then in the perifocal zone and in intact tissues. Conclusions: Regulation and stimulation of such changes were performed at early stages by autocrine regulatory mechanisms and cells of primary and growing melanoma, and later by the paracrine method spreading its influence on surrounding tissues and the whole body.

Aim: comparative analysis of uPA-Ag, uPA-act, tPA-Ag, tPA-act, PAI-1-Ag and PAI-1-act in tissues of esophageal squamous cell carcinoma (ESCC) in men (M) and women (W) to clarify some pathogenesis issues. Material and methods. Tumor tissue of ESCC and its perifocal area (19 M and 8 menopausal W aged 38 - 72 years, st II, G2, T2-3N0-1M0-1) were studied by ELISA using standard test kits. Results. ESCC tissue of M showed an increase in both uPA types from the resection line (RL), while in W only uPA-act was increased. tPA-Ag was decreased in tumors of W, tPA-act - in tumors of all patients. Levels of both PAI-1 types were higher in ESCC of M than in W. In M, tPA-Ag in perifocal area was lower than in RL, while in W, on the contrary, it was higher. tPA-Ag/tPA-act coefficient in M tumors was 5.7 times higher than in W; in perifocal area it was reduced in all patients, being at the same time in M lower than in W. Both PAI-1 types were increased in malignant tissues of all patients, prevailing in M tumors, and PAI-1-Ag in peritumoral tissue was higher in M than in W. Conclusions. Significant gender differences were found in expression of uPA, tPA and PAI-1 in tissues of esophageal squamous cell carcinoma. Presence of tumor-associated uPA, PAI-1 and tPA in men and uPA and PAI-1 in women is possible in tumor tissue of esophageal squamous cell carcinoma and its perifocal area. Determination of plasminogen regulation system in tissues of esophageal squamous cell carcinoma can be used for the selection and individualization of postoperative treatments.