Russian Journal of Biological Physics and Chemisrty Russian Journal of Biological Physics and Chemisrty АКТУАЛЬНЫЕ ВОПРОСЫ БИОЛОГИЧЕСКОЙ ФИЗИКИ И ХИМИИ 2499-9962 55118 10.29039/rusjbpc.2022.0540 МЕДИЦИНСКАЯ БИОФИЗИКА И БИОФИЗИЧЕСКАЯ ХИМИЯ MEDICAL BIOPHYSICS AND BIOPHYSICAL CHEMISTRY МЕДИЦИНСКАЯ БИОФИЗИКА И БИОФИЗИЧЕСКАЯ ХИМИЯ ANALYSIS OF MICROSOMAL REDUCTASE ACTIVITY IN OVARIAN TISSUE AFTER CRYOPRESERVATION BY ENHANCED CHEMILUMINESCENCE АНАЛИЗ АКТИВНОСТИ МИКРОСОМАЛЬНЫХ РЕДУКТАЗ ТКАНИ ЯИЧНИКОВ ПОСЛЕ КРИОКОНСЕРВАЦИИ МЕТОДОМ АКТИВИРОВАННОЙ ХЕМИЛЮМИНЕСЦЕНЦИИ Проскурнина Е. В. Proskurnina E. V. proskurnina@gmail.com Созарукова М. М. Sozarukova M. M. s_madinam@bk.ru Федорова М. В. Fedorova M. V. theklazontag@yandex.ru Киселева М. В. Kiseleva M. V. oxiscience@mail.ru Медико-генетический научный центр им. академика Н.П. Бочкова Москва Россия Research Centre for Medical Genetics Moscow Russian Federation Институт общей и неорганической химии им. Н.С. Курнакова РАН Москва Россия Kurnakov Institute of Gentral and Inorganic Chemistry of Russian Academy of Sciences Moscow Russian Federation Центральный НИИ эпидемиологии Роспотребнадзора Москва Россия Central Research Institute for Epidemiology» of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing Moscow Russian Federation Национальный медицинский исследовательский центр радиологии Минздрава России Москва Россия National Medical Research Radiological Centre of the Ministry of Health of the Russian Federation Moscow Russian Federation 28 09 2022 28 09 2022 7 3 434 439 20 09 2022 20 09 2022 https://rusjbpc.ru/en/nauka/article/55118/view

Цель исследования — изучить активность НАДН-зависимой цитохром b5-редуктазы (CYB5R) и НАДФН-зависимой цитохром P450-редуктазы (CYPOR) в тканях яичника после криоконсервации методом люцигенин-активированной хемилюминесценции со стимулами НАДН и НАДФН, соответственно. Результаты свидетельствуют о том, что в криоконсервированных тканях яичников сохраняется как активность митохондрий, так и микросомальных редуктаз. После криоконсервации уровень выработки митохондриями супероксидного анион-радикала падает в 3–10 раз, при этом наличие или отсутствие химиотерапии влияния не оказывает, также на этот параметр не оказывает влияния тяжесть заболевания. По сравнению с группой контроля (пациентки с доброкачественными новообразованиями), активность CYB5R и CYPOR ткани яичников при злокачественном онкологическом процессе уменьшается в 1,5–10 раз в зависимости от тяжести заболевания, и наличие химиотерапии резко влияет на эти показатели — при химиотерапии активность микросомальных редуктаз падает в 50–100 раз по сравнению с группой контроля. Таким образом, активность микросомальных редуктаз является более информативным параметром для оценки функциональности криоконсервированной ткани яичников, чем супероксид-продуцирующая способность митохондрий, поскольку, во-первых, зависит от стадии заболевания и предшествующей химиотерапии, и во-вторых, аналитический сигнал НАДН/НАДФН стимулированной хемилюминесценции характеризуется примерно в 30 раз более высокой интенсивностью, чем митохондриальная хемилюминесценция, что обуславливает более высокую аналитическую чувствительность методики.

The aim of the study was to investigate the activity of NADH-dependent cytochrome b5 reductase (CYB5R) and NADPH-dependent cytochrome P450 reductase (CYPOR) in ovarian tissues after cryopreservation by lucigenin-enhanced chemiluminescence with NADH and NADPH stimulation, respectively. The results indicate that both mitochondrial and microsomal reductase activities are preserved in cryopreserved ovarian tissues. After cryopreservation, the level of production of superoxide anion radical by mitochondria drops by 3–10 times, while the presence or absence of chemotherapy has no effect, and this parameter is also not affected by the severity of the disease. Compared to the control group (patients with benign tumors), the activity of CYB5R and CYPOR of ovarian tissue in a malignant cancer process decreases 1.5–10 times depending on the disease severity, and the presence of chemotherapy dramatically affects these parameters — the activity of microsomal reductases decreases by 50–100 times in chemotherapy compared to the control group. Thus, microsomal reductase activity is a more informative parameter for assessing the functionality of cryopreserved ovarian tissue than superoxide-producing capacity of mitochondria, because, firstly, it depends on the stage of disease and prior chemotherapy, and secondly, the analytical signal of NADH/NADPH stimulated chemiluminescence is characterized by approximately 30 times higher intensity than mitochondrial chemiluminescence, which leads to higher analytical sensitivity of the technique.

 цитохром b5-редуктаза цитохром Р450-редуктаза яичники криоконсервация хемилюминесценция cytochrome b5 reductase cytochrome P450 reductase ovaries cryopreservation chemiluminescence

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