Hunter Arwen L, Choy Jonathan C, Granville David J
Department of Pathology and Laboratory Medicine, St. Paul's Hospital, University of British Columbia, Vancouver, Canada.
Methods Mol Med. 2005;112:277-89. doi: 10.1385/1-59259-879-x:277.
The past decade has seen a surge in research devoted to understanding the role of cell death in the pathogenesis of various forms of cardiovascular disease. In particular, apoptosis has received much attention owing to the tightly regulated biochemical nature of this form of cell death and the realization of potential therapeutic opportunities. The current chapter describes a few of the more widely used protocols for detecting and quantifying apoptosis in cardiovascular tissues. Specifically, this chapter describes terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining for DNA fragmentation, Hoechst staining for chromatin condensation, annexin V labeling of phosphatidylserine externalization, and Western blot and immunoflorescence detection of caspase cleavage and activation, respectively.
在过去十年中,致力于了解细胞死亡在各种心血管疾病发病机制中作用的研究激增。特别是,由于这种细胞死亡形式具有严格调控的生化特性以及潜在治疗机会的实现,细胞凋亡受到了广泛关注。本章介绍了一些在心血管组织中检测和定量细胞凋亡的更广泛使用的方法。具体而言,本章分别描述了用于检测DNA片段化的末端脱氧核苷酸转移酶dUTP缺口末端标记(TUNEL)染色、用于检测染色质凝聚的Hoechst染色、用于检测磷脂酰丝氨酸外翻的膜联蛋白V标记,以及用于检测半胱天冬酶切割和激活的蛋白质免疫印迹法和免疫荧光法。