Department of Internal Medicine, Osaka Dental University, 8-1 Kuzuhahanazono-cho, Hirakata, Osaka 573-1121, Japan.
Biochem Biophys Res Commun. 2011 Aug 5;411(3):569-73. doi: 10.1016/j.bbrc.2011.06.186. Epub 2011 Jul 6.
Apoptotic cell death eventually results in secondary necrotic cell death, whereas caspase-independent primary necrotic cell death has been reported and its mechanism involving RIP1 and RIP3 has been recently elucidated. Dual staining with fluorescent Annexin V and propidium iodide (PI) has been used to discriminate apoptotic and necrotic cell death, in which Annexin V-positive/PI-negative staining is regarded as apoptosis and PI-positive staining as necrosis. Here we demonstrate that primary necrotic cells unexpectedly show Annexin V-positive/PI-negative staining before they become PI-positive, and that primary necrotic and apoptotic Annexin V-positive/PI-negative cells can be discriminated by necrostatin-1, an inhibitor of primary necrosis by inhibition of RIP1.
细胞凋亡最终导致继发性坏死细胞死亡，而细胞凋亡以外的非经典途径的原发性坏死细胞死亡已经被报道，其涉及 RIP1 和 RIP3 的机制最近已经阐明。使用荧光 Annexin V 和碘化丙啶 (PI) 的双重染色已被用于区分凋亡和坏死细胞死亡，其中 Annexin V 阳性/PI 阴性染色被认为是凋亡，而 PI 阳性染色则被认为是坏死。在这里，我们证明原发性坏死细胞在变为 PI 阳性之前，出乎意料地表现出 Annexin V 阳性/PI 阴性染色，并且可以通过抑制 RIP1 的坏死抑制剂 necrostatin-1 来区分原发性坏死和凋亡 Annexin V 阳性/PI 阴性细胞。
