Department of Pharmacology and Toxicology, School of Pharmacy, Ardabil University of Medical Sciences, Ardabil, Iran.
Traditional Medicine and Hydrotherapy Research Center, Ardabil University of Medical Sciences, Ardabil, Iran.
Methods Mol Biol. 2024;2753:231-249. doi: 10.1007/978-1-0716-3625-1_11.
Peripheral blood lymphocytes as primary cells can be isolated from human, animal, fetus, and placenta. These cells are an excellent cellular model for the assessment of cytotoxicity, genotoxicity, oxidative stress, and mitochondrial and lysosomal dysfunction induced by drug and chemicals. Moreover, peripheral blood lymphocytes are an easily available source of primary cells appropriate for basic research and in cellular studies regarding teratogenic, genotoxic, and cytotoxic effect of drugs and chemicals. Most drugs and other chemicals that produce birth defects, known as teratogenic agents, produce reactive oxygen species (ROS) formation and mitochondrial and lysosomal dysfunction. It seems that there is an important mechanistic link between oxidative stress, mitochondrial damages, lysosomal integrity, and teratogenic drug-induced birth defects. One of the most sensitive periods in the embryo is transition from an important developmental event to another such as transition from proliferation to differentiation. Mitochondria, lysosomes, and cellular ROS have an important role in proliferative, differentiative, and apoptotic activities during the development. Therefore, disruption of the function of mitochondria, lysosomes, oxidative stress, and redox imbalance leads to cellular dysfunctions and subsequently poor developmental outcomes in the fetus. In this chapter, we will focus on evaluation of mitochondrial/lysosomal functions and estimation of ROS formation using flow cytometry methods in isolated lymphocytes and their isolated mitochondria.
外周血淋巴细胞作为原代细胞可从人、动物、胎儿和胎盘分离得到。这些细胞是评估药物和化学物质引起的细胞毒性、遗传毒性、氧化应激以及线粒体和溶酶体功能障碍的极佳细胞模型。此外，外周血淋巴细胞是一种易于获得的原代细胞来源，适用于基础研究和细胞研究，包括药物和化学物质的致畸、遗传毒性和细胞毒性作用。大多数导致出生缺陷的药物和其他化学物质，称为致畸剂，会产生活性氧（ROS）形成和线粒体和溶酶体功能障碍。氧化应激、线粒体损伤、溶酶体完整性和致畸药物引起的出生缺陷之间似乎存在重要的机制联系。胚胎最敏感的时期之一是从一个重要的发育事件过渡到另一个事件，例如从增殖到分化的过渡。线粒体、溶酶体和细胞内 ROS 在发育过程中的增殖、分化和凋亡活动中发挥重要作用。因此，线粒体、溶酶体功能障碍、氧化应激和氧化还原失衡会导致细胞功能障碍，随后导致胎儿发育不良。在本章中，我们将重点关注使用流式细胞术方法在外周血淋巴细胞及其分离的线粒体中评估线粒体/溶酶体功能和估计 ROS 形成。
