Vinokurov A Yu, Popov S V, Belyakov D Yu, Popov D Yu, Nikulin A S, Zakrzhevskaya V D, Guseinov R G, Sivak K V, Dunaev A V, Potapova E V, Abramov A Yu
PhD, Senior Researcher, Research and Development Center of Biomedical Photonics; Orel State University, 95 Komsomolskaya St., Orel, 302026, Russia.
MD, DSc, Professor, Chief Physician; Clinical Hospital of St. Luke, 46 letter A, Chugunnaya St., Saint Petersburg, 194044, Russia; Head of the City Center for Endoscopic Urology and New Technologies; Clinical Hospital of St. Luke, 46 letter A, Chugunnaya St., Saint Petersburg, 194044, Russia; Professor, Department of Urology; Military Medical Academy, 6 letter Zh, Akademika Lebedeva St., Saint Petersburg, 194044, Russia; Head of the Department of Surgery and Urology; Saint Petersburg Medical and Social Institute, 72 letter A, Kondratyevsky Prospekt, Saint Petersburg, 195271, Russia.
Sovrem Tekhnologii Med. 2025;17(1):93-106. doi: 10.17691/stm2025.17.1.09. Epub 2025 Feb 28.
Hypoxia is a part of many pathological and some physiological processes. It also occurs as a result of surgical techniques associated with limiting the blood supply to the operated organs and tissues. Hypoxia leads to a significant decrease in the ability of cells to implement energy-dependent processes due to a reduced contribution of mitochondria to the synthesis of adenosine triphosphate (ATP). In order to protect cells and increase the time of surgery, infusion of a solution of sodium fumarate for several days before the surgical procedure is suggested. However, the mechanism of the observed protective effect is still a subject of discussion. the mechanism of the sodium fumarate cytoprotective effect on renal epithelial cells in acute hypoxia modeling by reducing oxygen in the medium using sodium dithionite.
The study was conducted using the MDCK renal epithelial cell line with sodium dithionite at a concentration of 5 mM to create hypoxic conditions. The parameters of cellular metabolism (including the value of mitochondrial membrane potential, the state of mitochondrial NADH and FAD, the content of Ca and Mg and the pH level in the cytosol, the rate of glucose absorption by cells, and cell death) were assessed by means of confocal and wide-field fluorescence microscopy. The concentration of dissolved oxygen was established using the polarographic method with a Clark electrode.
It was demonstrated that the use of sodium dithionite allows modeling acute hypoxia with a rapid decrease in the oxygen concentration in the cell incubation medium, which resulted in a change in mitochondrial function and the apoptosis progression. At that, sodium fumarate reduces the level of cell death, which is associated not with the restoration of the ATP-producing ability of mitochondria, but rather with an increase in the contribution of alternative sources of high-energy compounds.
At the cellular level, using an optimized hypoxia model, the study revealed the mechanism of the protective role of sodium fumarate, which explained the antihypoxant effectiveness in assisted ischemia of organs and tissues.
缺氧是许多病理过程和一些生理过程的一部分。它也可能由于与限制手术器官和组织血液供应相关的手术技术而出现。缺氧会导致细胞进行能量依赖过程的能力显著下降，这是因为线粒体对三磷酸腺苷（ATP）合成的贡献减少。为了保护细胞并延长手术时间，建议在手术前几天输注富马酸钠溶液。然而，观察到的保护作用机制仍是一个讨论的话题。通过连二亚硫酸钠降低培养基中的氧气含量，研究富马酸钠对急性缺氧模型中肾上皮细胞的细胞保护作用机制。
本研究使用MDCK肾上皮细胞系，用浓度为5 mM的连二亚硫酸钠来制造缺氧条件。通过共聚焦和宽视野荧光显微镜评估细胞代谢参数（包括线粒体膜电位值、线粒体NADH和FAD状态、钙和镁含量以及细胞质中的pH水平、细胞对葡萄糖的吸收速率和细胞死亡情况）。使用带有克拉克电极的极谱法测定溶解氧浓度。
结果表明，使用连二亚硫酸钠能够模拟急性缺氧，使细胞培养液中的氧浓度迅速降低，这导致线粒体功能改变和细胞凋亡进程。此时，富马酸钠降低了细胞死亡水平，这与线粒体产生ATP能力的恢复无关，而是与高能化合物替代来源的贡献增加有关。
在细胞水平上，通过使用优化的缺氧模型，该研究揭示了富马酸钠的保护作用机制，这解释了其在器官和组织辅助缺血中的抗缺氧有效性。
