炎症体
钾通道
化学
钾
细胞生物学
生物物理学
生物
生物化学
受体
有机化学
作者
C.N. Immanuel,Bo Teng,Bo Dong,Elizabeth M. Gordon,Charlean Luellen,Benjamín López,Jeffrey N Harding,Stephania A. Cormier,Elizabeth Fitzpatrick,Andreas Schwingshackl,Christopher M. Waters
出处
期刊:American Journal of Physiology-lung Cellular and Molecular Physiology
[American Physiological Society]
日期:2024-01-22
标识
DOI:10.1152/ajplung.00313.2023
摘要
Because of the importance of potassium efflux in inflammasome activation, we investigated the role of the two-pore potassium (K2P) channel TREK-1 in macrophage inflammasome activity. Using primary alveolar macrophages (AMs) and bone marrow derived macrophages (BMDMs) from wild type (wt) and TREK-1-/- mice, we measured responses to inflammasome priming (using LPS) and activation (LPS +ATP). We measured IL-1β, caspase-1, and NLRP3 via ELISA and western blot. A membrane-permeable potassium indicator was used to measure potassium efflux during ATP exposure, and a fluorescence-based assay was used to assess changes in membrane potential. Inflammasome activation induced by LPS+ATP increased IL-1β secretion from wt AMs, whereas activation was significantly reduced in TREK-1-/- AMs. Priming of BMDMs using LPS was not affected by either genetic deficiency or pharmacological inhibition of TREK-1 with Spadin. Cleavage of caspase-1 following LPS+ATP treatment was significantly reduced in TREK-1-/- BMDMs. The intracellular potassium concentration in LPS primed wt BMDMs was significantly lower compared with TREK-1-/- BMDMs or wt BMDMs treated with Spadin. Conversely, activation of TREK-1 with BL1249 caused a decrease in intracellular potassium in wt BMDMs. Treatment of LPS-primed BMDMs with ATP caused a rapid reduction in intracellular potassium levels, with the largest change observed in TREK-1-/- BMDMs. Intracellular K+ changes were associated with changes in the plasma membrane potential (Em), as evidenced by a more depolarized Em in TREK-1-/- BMDMs compared with wt, and Em hyperpolarization upon TREK-1 channel opening with BL1249. These results suggest that TREK-1 is an important regulator of NLRP3 inflammasome activation in macrophages.
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