Khotina Victoria A, Vinokurov Andrey Y, Sinyov Vasily V, Zhuravlev Alexander D, Popov Daniil Y, Sukhorukov Vasily N, Sobenin Igor A, Orekhov Alexander N
Institute of General Pathology and Pathophysiology, 8 Baltiyskaya Street, 125315Moscow, Russia.
Cell Physiology and Pathology Laboratory of R&D Center of Biomedical Photonics, Orel State University, 95 Komsomolskaya Street, 302026 Orel, Russia.
Curr Med Chem. 2024 Oct 11. doi: 10.2174/0109298673323639240926095549.
Atherosclerosis is a complex cardiovascular disease often associated with mitochondrial dysfunction, which can lead to various cellular and metabolic abnormalities. Within the mitochondrial genome, specific mutations have been implicated in contributing to mitochondrial dysfunction. Atherosclerosis-associated m.15059G>A mutation has been of particular interest due to its potential role in altering mitochondrial function and cellular health.
This study aims to investigate the role of the atherosclerosis-associated m.15059G>A mutation in the development of mitochondrial dysfunction in monocyte-- like cells.
Monocyte-like cytoplasmic hybrid cell line TC-HSMAM1, which contains the m.15059G>A mutation in mtDNA, was used. The MitoCas9 vector was utilized to eliminate mtDNA copies carrying the m.15059G>A mutation from TC-HSMAM1 cybrids. Mitochondrial membrane potential, generation of reactive oxygen species, and lipid peroxidation levels were assessed using flow cytometry. Cellular reduced glutathione levels were assessed using the confocal microscopy. The oxygen consumption rate was measured using polarographic oxygen respirometry.
The elimination of the m.15059G>A mutation resulted in a significant increase in mitochondrial membrane potential and improved mitochondrial efficiency while also causing a decrease in the generation of reactive oxygen species, lipid peroxidation, as well as cellular bioenergetic parameters, such as proton leak and non-mitochondrial oxygen consumption. At the same time, no changes were found in the intracellular antioxidant system after the mitochondrial genome editing.
The presence of the m.15059G>A mutation contributes to mitochondrial dysfunction by reducing mitochondrial membrane potential, increasing the generation of reactive oxygen species and lipid peroxidation, and altering mitochondrial bioenergetics. Elimination of the mtDNA containing atherogenic mutation leads to an improvement in mitochondrial function.
动脉粥样硬化是一种复杂的心血管疾病，常与线粒体功能障碍相关，可导致各种细胞和代谢异常。在线粒体基因组中，特定突变被认为与线粒体功能障碍有关。与动脉粥样硬化相关的m.15059G>A突变因其在改变线粒体功能和细胞健康方面的潜在作用而备受关注。
本研究旨在探讨与动脉粥样硬化相关的m.15059G>A突变在单核细胞样细胞线粒体功能障碍发生发展中的作用。
使用单核细胞样细胞质杂交细胞系TC-HSMAM1，其线粒体DNA中含有m.15059G>A突变。利用MitoCas9载体从TC-HSMAM1细胞质杂种中消除携带m.15059G>A突变的线粒体DNA拷贝。使用流式细胞术评估线粒体膜电位、活性氧生成和脂质过氧化水平。使用共聚焦显微镜评估细胞内还原型谷胱甘肽水平。使用极谱氧呼吸测定法测量氧消耗率。
消除m.15059G>A突变导致线粒体膜电位显著增加，线粒体效率提高，同时活性氧生成、脂质过氧化以及细胞生物能量参数（如质子泄漏和非线粒体氧消耗）减少。同时，线粒体基因组编辑后细胞内抗氧化系统未发现变化。
m.15059G>A突变的存在通过降低线粒体膜电位、增加活性氧生成和脂质过氧化以及改变线粒体生物能量学导致线粒体功能障碍。消除含有致动脉粥样硬化突变的线粒体DNA可改善线粒体功能。
