Academy of Medical Engineering and Translational Medicine, Tianjin University, Tianjin, China.
Tsinghua Institute for Immunology and Department of Basic Medical Sciences, Beijing Key Lab for Immunological Research on Chronic Diseases, School of Medicine, Tsinghua University, Beijing, China.
Methods Mol Biol. 2022;2459:85-92. doi: 10.1007/978-1-0716-2144-8_9.
Potassium ion (K) efflux is often considered as an upstream signaling event of NLRP3 activation. The main evidence to demonstrate the importance of K efflux is that high concentration of extracellular K inhibits NLRP3 inflammasome assembly. However, the conditions used to prevent K flowing also breaks down a basic parameter of eukaryotic biology, leading to sustained membrane potential depolarization and affecting normal signal transduction in cells. Therefore, direct measurement of intracellular ion concentration can more truly reflect the role of K flow during the activation of NLRP3. In this chapter, we will provide the rationale and a method to evaluate intracellular K concentration by ICP-OES (Inductively Coupled Plasma Optical Emission Spectroscopy), which helps us understand how disturbances in intracellular K level orchestrates NLRP3 inflammasome activation.
钾离子(K)外流通常被认为是 NLRP3 激活的上游信号事件。证明 K 外流重要性的主要证据是,细胞外高浓度的 K 抑制 NLRP3 炎性体的组装。然而,用于阻止 K 流动的条件也破坏了真核生物生物学的一个基本参数,导致持续的膜电位去极化,并影响细胞内的正常信号转导。因此,直接测量细胞内离子浓度可以更真实地反映 NLRP3 激活过程中 K 流动的作用。在本章中,我们将提供通过 ICP-OES(电感耦合等离子体光学发射光谱)评估细胞内 K 浓度的原理和方法,这有助于我们了解细胞内 K 水平的紊乱如何协调 NLRP3 炎性体的激活。