通过测量释放的乳酸脱氢酶活性来检测细胞焦亡。
Detection of pyroptosis by measuring released lactate dehydrogenase activity.
作者信息
Rayamajhi Manira, Zhang Yue, Miao Edward A
机构信息
Department of Microbiology and Immunology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
出版信息
Methods Mol Biol. 2013;1040:85-90. doi: 10.1007/978-1-62703-523-1_7.
Abstract
Pyroptosis is a form of programmed, inflammatory cell death that is dependent on the activation of a cysteine protease caspase-1. Following caspase-1 activation via inflammasomes (including NLRP3, NLRC4, Nlrp1b, and AIM2), cells lose membrane integrity and lyse, releasing lactate dehydrogenase (LDH) that is normally maintained within the cell cytosol. Thus, pyroptosis is distinct from apoptosis, which results in cellular contents being enclosed within membrane blebs during cellular demise. LDH is only released from apoptotic blebs after secondary necrosis occurs. Pyroptosis is distinct from necrosis in that it requires the activity of caspase-1. In this chapter, we describe enzymatic assays for the detection of LDH released by pyroptotic cells using a commercially available kit, as well as a simple and cost-effective method adapted from Decker et al. (J Immunol Methods 115:61-69, 1988).
摘要
细胞焦亡是一种程序性炎症细胞死亡形式，依赖于半胱氨酸蛋白酶caspase-1的激活。通过炎性小体（包括NLRP3、NLRC4、Nlrp1b和AIM2）激活caspase-1后，细胞失去膜完整性并裂解，释放通常维持在细胞质中的乳酸脱氢酶（LDH）。因此，细胞焦亡不同于凋亡，凋亡导致细胞死亡期间细胞内容物被包裹在膜泡中。只有在继发性坏死发生后，LDH才会从凋亡小泡中释放出来。细胞焦亡与坏死不同，因为它需要caspase-1的活性。在本章中，我们描述了使用市售试剂盒检测细胞焦亡释放的LDH的酶促测定方法，以及一种改编自Decker等人（《免疫学方法杂志》115:61-69，1988年）的简单且经济高效的方法。
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