NLR family pyrin domain containing 3 (NLRP3) and caspase 1 (CASP1) modulation by intracellular Cl– concentration
Abstract: 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-1β (IL-1β), 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-1β mRNA expression and partially the IL-1β 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-1β mRNA response to Cl– and the IL-1β secretion, interrupting the autocrine IL-1β loop. The results suggest that Cl– effects are mediated by SGK1, in which under Cl– modulation stimulates the secretion of mature IL-1β, in turn, responsible for the upregulation of ROS, CASP1, NLRP3 and IL-1β itself, through autocrine signalling.
Main Authors: | , , , , , , , , , |
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Format: | Artículo biblioteca |
Language: | eng |
Published: |
Wiley-Blackwell
2021
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Subjects: | FIBROSIS QUISTICA REGULADOR DE LA CONDUCTANCIA TRANSMEMBRANA, REGULADOR DE CONDUCTANCIA TRANSMEMBRANA DE LA FIBROSIS QUÍSTICA, SEÑALIZACION AUTOCRINA, ANIÓN CLORURO, CANAL DE CLORURO, INFLAMASOMA, INTERLEUCINA, NLRP3, SEGUNDO MENSAJERO, SGK1, |
Online Access: | https://repositorio.uca.edu.ar/handle/123456789/14599 |
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Summary: | Abstract:
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-1β (IL-1β), 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-1β mRNA expression and partially the IL-1β 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-1β mRNA response
to Cl–
and the IL-1β secretion, interrupting the autocrine IL-1β loop. The results suggest that Cl–
effects are mediated by SGK1, in which under Cl–
modulation stimulates
the secretion of mature IL-1β, in turn, responsible for the upregulation of ROS, CASP1,
NLRP3 and IL-1β itself, through autocrine signalling. |
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