Figg Nichola L, Bennett Martin R
Division of Cardiovascular Medicine, Addenbrooke's Centre for Clinical Investigation, University of Cambridge, Box 110, Hills Road, Cambridge, CB2 0QQ, UK.
Methods Mol Biol. 2015;1339:191-9. doi: 10.1007/978-1-4939-2929-0_13.
Apoptosis is a key process occurring in atherosclerosis, both in humans and in animal models. Apoptosis occurs in all cell types studied thus far, and thus lineage marking is often necessary. Apoptosis should be ascertained using a combination of morphological features and activation of specific pathways (e.g., terminal UTP nick end labeling-TUNEL). Both TUNEL and cryptic epitope antibodies (e.g., cleaved caspase 3) can be used, although they will often give different frequencies. Apoptotic frequency but not rate can be estimated from these methods, as we do not know the timing of apoptosis or how much of the process is marked by each method. We describe the morphological and immunohistochemical methods used in our laboratory to detect apoptotic cells in animal and human atherosclerotic plaques.
细胞凋亡是人类和动物模型中动脉粥样硬化发生过程中的一个关键过程。迄今为止,在所有研究的细胞类型中均会发生细胞凋亡,因此谱系标记通常是必要的。细胞凋亡应通过形态学特征和特定途径的激活(例如,末端脱氧核苷酸转移酶介导的缺口末端标记法-TUNEL)相结合来确定。TUNEL法和隐蔽表位抗体(例如,裂解的半胱天冬酶3)均可使用,尽管它们给出的频率通常会有所不同。通过这些方法可以估计细胞凋亡的频率而非速率,因为我们不知道细胞凋亡的时间,也不知道每种方法标记了该过程的多少部分。我们描述了我们实验室用于检测动物和人类动脉粥样硬化斑块中凋亡细胞的形态学和免疫组织化学方法。