DAMP Laboratory, The Third Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China.
Department of Surgery, UT Southwestern Medical Center, Dallas, TX, USA.
Methods Mol Biol. 2023;2712:103-115. doi: 10.1007/978-1-0716-3433-2_10.
Ferroptosis is a type of regulated necrosis driven by uncontrolled membrane lipid peroxidation. Mitochondria, which are membrane-bound organelles present in almost all eukaryotic cells and play a central role in energy metabolism and various types of cell death, have a complicated role in ferroptosis. On one hand, mitochondrial-derived iron metabolism and reactive oxygen species (ROS) production may promote ferroptosis. On the other hand, mitochondria also possess a dihydroorotate dehydrogenase (DHODH)-dependent antioxidant system that detoxifies lipid peroxides. This chapter summarizes several methods, such as western blotting, immunofluorescence, cell viability assays, mitochondrial fluorescent probes, adenosine 5'-triphosphate (ATP) assay kits, mitochondrial respiration, and mitophagy tests, that may enable researchers to gain a deeper understanding of the dual role of mitochondria in ferroptosis.
铁死亡是一种由不受控制的膜脂质过氧化驱动的调节性细胞坏死。线粒体是一种存在于几乎所有真核细胞中的膜结合细胞器，在能量代谢和各种类型的细胞死亡中发挥核心作用，在线粒体铁死亡中具有复杂的作用。一方面，线粒体来源的铁代谢和活性氧（ROS）的产生可能会促进铁死亡。另一方面，线粒体还具有一种二氢乳清酸脱氢酶（DHODH）依赖性抗氧化系统，可解毒脂质过氧化物。本章总结了几种方法，如western blotting、免疫荧光、细胞活力测定、线粒体荧光探针、三磷酸腺苷（ATP）测定试剂盒、线粒体呼吸和噬线粒体试验，这些方法可以使研究人员更深入地了解线粒体在铁死亡中的双重作用。
