Medical Faculty, Institute of Pharmacology and Experimental Toxicology, University of Ljubljana, Ljubljana, Slovenia.
Drug Chem Toxicol. 2012 Oct;35(4):399-405. doi: 10.3109/01480545.2011.633087. Epub 2012 Feb 29.
Apoptosis and necroptosis are highly regulated, interconnected forms of a cell death. The distinction between them is critical, because necroptosis may cause significant cell loss and local inflammation, whereas apoptosis is essential for tissue homeostasis. The same stimulus can induce both apoptosis and necroptosis. Both forms of a cell death were detected in various pathologies, including pathologies in the central nervous system. Astrocytes are a large, heterogeneous cell population in the central nervous system, with many supportive, developmental functions. Although their demise may seriously impair normal functions of the central nervous system, it is still poorly understood. In this study, apoptosis and necroptosis were induced in cultured rat astrocytes by staurosporine. When a low concentration (10(-7) M) of staurosporine was applied, a significantly increased proportion of early apoptotic cells was detected after regeneration in a staurosporine free medium. The proportion of necroptotic cells was already increased without regeneration after 3 hours of exposure to staurosporine. When a higher (10(-6) M) concentration of staurosporine was applied, further significantly increased necroptosis was detected after regeneration in a staurosporine free medium. Necroptosis was significantly reduced when RIP1 kinase was inhibited by necrostatin-1, whereas inhibition of caspases with z-vad-fmk, an irreversible pan-caspase inhibitor, did not prevent necroptosis. This report of necroptosis induced by staurosporine represents a simple approach for the in vitro induction and detection of apoptosis and necroptosis.
细胞凋亡和坏死性细胞凋亡是高度调控的、相互关联的细胞死亡形式。它们之间的区别是至关重要的，因为坏死性细胞凋亡可能导致大量细胞损失和局部炎症，而细胞凋亡是组织稳态所必需的。相同的刺激可以诱导细胞凋亡和坏死性细胞凋亡。这两种形式的细胞死亡都在各种病理学中被检测到，包括中枢神经系统的病理学。星形胶质细胞是中枢神经系统中一种大型的、异质性的细胞群体，具有许多支持性的、发育性的功能。尽管它们的死亡可能严重损害中枢神经系统的正常功能，但目前对此仍知之甚少。在这项研究中，星形胶质细胞在培养的大鼠星形胶质细胞中通过星形孢菌素诱导凋亡和坏死性细胞凋亡。当应用低浓度（10(-7) M）的星形孢菌素时，在无星形孢菌素的培养基中再生后，早期凋亡细胞的比例显著增加。在暴露于星形孢菌素 3 小时后，没有再生，坏死性细胞的比例已经增加。当应用较高浓度（10(-6) M）的星形孢菌素时，在无星形孢菌素的培养基中再生后，进一步检测到明显增加的坏死性细胞凋亡。当 RIP1 激酶被坏死素-1抑制时，坏死性细胞凋亡显著减少，而用不可逆的多半胱天冬酶抑制剂 z-vad-fmk 抑制半胱天冬酶并不能防止坏死性细胞凋亡。本报告中星形孢菌素诱导的坏死性细胞凋亡代表了一种简单的体外诱导和检测细胞凋亡和坏死性细胞凋亡的方法。
