期刊
IMMUNOLOGY
卷 163, 期 4, 页码 493-511出版社
WILEY
DOI: 10.1111/imm.13336
关键词
autocrine signalling; CFTR; chloride anion; chloride channel; inflammasome; interleukin; NLRP3; ROS; second messenger; SGK1
类别
资金
- National Agency for the Promotion of Science and Technology (ANPCYT) [PICT 2012-1278, PICT 2018-04429]
- National Scientific and Technical Research Council of Argentina (CONICET) [PIP 2015-2017 11220150100227CO, PUE 2016 22920160100129CO]
- Pontifical Catholic University of Argentina (UCA)
- Pablo Cassara Foundation
- CONICET
The modulation of intracellular chloride (Cl-) concentration indirectly affects the activity of NLRP3 and CASP1, potentially through SGK1 mediation. The autocrine positive feedback loop involves multiple signaling molecules and pathways that regulate the secretion of IL-1 beta.
The impairment of the cystic fibrosis transmembrane conductance regulator (CFTR) activity induces intracellular chloride (Cl-) accumulation. The anion Cl-, acting as a second messenger, stimulates the secretion of interleukin-1 beta (IL-1 beta), which starts an autocrine positive feedback loop. Here, we show that NLR family pyrin domain containing 3 (NLRP3) and caspase 1 (CASP1) are indirectly modulated by the intracellular Cl- concentration, showing maximal expression and activity at 75 mM Cl-, in the presence of the ionophores nigericin and tributyltin. The expression of PYD and CARD domain containing (PYCARD/ASC) remained constant from 0 to 125 mM Cl-. The CASP1 inhibitor VX-765 and the NLRP3 inflammasome inhibitor MCC950 completely blocked the Cl--stimulated IL-1 beta mRNA expression and partially the IL-1 beta secretion. DCF fluorescence (cellular reactive oxygen species, cROS) and MitoSOX fluorescence (mitochondrial ROS, mtROS) also showed maximal ROS levels at 75 mM Cl-, a response strongly inhibited by the ROS scavenger N-acetyl-L-cysteine (NAC) or the NADPH oxidase (NOX) inhibitor GKT137831. These inhibitors also affected CASP1 and NLRP3 mRNA and protein expression. More importantly, the serum/glucocorticoid regulated kinase 1 (SGK1) inhibitor GSK650394, or its shRNAs, completely abrogated the IL-1 beta mRNA response to Cl- and the IL-1 beta secretion, interrupting the autocrine IL-1 beta loop. The results suggest that Cl- effects are mediated by SGK1, in which under Cl- modulation stimulates the secretion of mature IL-1 beta, in turn, responsible for the upregulation of ROS, CASP1, NLRP3 and IL-1 beta itself, through autocrine signalling.
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