Nuclear Factor-κB Signaling Regulates the Nociceptin Receptor but Not Nociceptin Itself

The nociceptin receptor (NOP) and nociceptin are involved in the pathways of pain and inflammation. The potent role of nuclear factor-κB (NFκB) in the modulation of tumor necrosis factor-α (TNF-α) and interleukin (IL)-1β on the nociceptin system in human THP-1 cells under inflammatory conditions wer...

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Main Authors: Lan Zhang, Ulrike M. Stamer, Robin Moolan-Vadackumchery, Frank Stüber
Format: Article
Language:English
Published: MDPI AG 2024-12-01
Series:Cells
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Online Access:https://www.mdpi.com/2073-4409/13/24/2111
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author Lan Zhang
Ulrike M. Stamer
Robin Moolan-Vadackumchery
Frank Stüber
author_facet Lan Zhang
Ulrike M. Stamer
Robin Moolan-Vadackumchery
Frank Stüber
author_sort Lan Zhang
collection DOAJ
description The nociceptin receptor (NOP) and nociceptin are involved in the pathways of pain and inflammation. The potent role of nuclear factor-κB (NFκB) in the modulation of tumor necrosis factor-α (TNF-α) and interleukin (IL)-1β on the nociceptin system in human THP-1 cells under inflammatory conditions were investigated. Cells were stimulated without/with phorbol-myristate-acetate (PMA), TNF-α, IL-1β, or PMA combined with individual cytokines. To examine NFκB’s contribution to the regulation of the nociceptin system, PMA-stimulated cells were treated with NFκB inhibitor BAY 11-7082, JSH-23, or anacardic acid before culturing with TNF-α or IL-1β. <i>NOP</i> and prepronociceptin (<i>ppNOC</i>) mRNA were quantified by RT-qPCR; cell membrane NOP and intracellular nociceptin protein levels were measured by flow cytometry. Phosphorylation and localization of NFκB/p65 were determined using ImageStream. PMA + TNF-α decreased <i>NOP</i> mRNA compared to stimulation with PMA alone, while PMA + IL-1β did not. BAY 11-7082 and JSH-23 reversed the repression of <i>NOP</i> by PMA + TNF-α. TNF-α and IL-1β attenuated PMA’s upregulating effects on <i>ppNOC</i>. None of the inhibitors preserved the upregulation of <i>ppNOC</i> in PMA + TNF-α and PMA + IL-1β cultures. TNF-α strongly mediated the nuclear translocation of NFκB/p65 in PMA-treated cells, while IL-1β did not. Proinflammatory cytokines suppressed <i>NOP</i> and <i>ppNOC</i> mRNA in PMA-induced human THP-1 cells. NFκB signaling seems to be an important regulator controlling the transcription of NOP. These findings suggest that the nociceptin system may play an anti-inflammatory role during immune responses.
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spelling doaj-art-5fcc68da26474db083effc0c687f1cc32024-12-27T14:16:38ZengMDPI AGCells2073-44092024-12-011324211110.3390/cells13242111Nuclear Factor-κB Signaling Regulates the Nociceptin Receptor but Not Nociceptin ItselfLan Zhang0Ulrike M. Stamer1Robin Moolan-Vadackumchery2Frank Stüber3Department of Anaesthesiology and Pain Medicine, Inselspital, Bern University Hospital, University of Bern, 3010 Bern, SwitzerlandDepartment of Anaesthesiology and Pain Medicine, Inselspital, Bern University Hospital, University of Bern, 3010 Bern, SwitzerlandDepartment of Anaesthesiology and Pain Medicine, Inselspital, Bern University Hospital, University of Bern, 3010 Bern, SwitzerlandDepartment of Anaesthesiology and Pain Medicine, Inselspital, Bern University Hospital, University of Bern, 3010 Bern, SwitzerlandThe nociceptin receptor (NOP) and nociceptin are involved in the pathways of pain and inflammation. The potent role of nuclear factor-κB (NFκB) in the modulation of tumor necrosis factor-α (TNF-α) and interleukin (IL)-1β on the nociceptin system in human THP-1 cells under inflammatory conditions were investigated. Cells were stimulated without/with phorbol-myristate-acetate (PMA), TNF-α, IL-1β, or PMA combined with individual cytokines. To examine NFκB’s contribution to the regulation of the nociceptin system, PMA-stimulated cells were treated with NFκB inhibitor BAY 11-7082, JSH-23, or anacardic acid before culturing with TNF-α or IL-1β. <i>NOP</i> and prepronociceptin (<i>ppNOC</i>) mRNA were quantified by RT-qPCR; cell membrane NOP and intracellular nociceptin protein levels were measured by flow cytometry. Phosphorylation and localization of NFκB/p65 were determined using ImageStream. PMA + TNF-α decreased <i>NOP</i> mRNA compared to stimulation with PMA alone, while PMA + IL-1β did not. BAY 11-7082 and JSH-23 reversed the repression of <i>NOP</i> by PMA + TNF-α. TNF-α and IL-1β attenuated PMA’s upregulating effects on <i>ppNOC</i>. None of the inhibitors preserved the upregulation of <i>ppNOC</i> in PMA + TNF-α and PMA + IL-1β cultures. TNF-α strongly mediated the nuclear translocation of NFκB/p65 in PMA-treated cells, while IL-1β did not. Proinflammatory cytokines suppressed <i>NOP</i> and <i>ppNOC</i> mRNA in PMA-induced human THP-1 cells. NFκB signaling seems to be an important regulator controlling the transcription of NOP. These findings suggest that the nociceptin system may play an anti-inflammatory role during immune responses.https://www.mdpi.com/2073-4409/13/24/2111cell culturescytokinesNFκBnociceptinnociceptin receptorsignal transduction
spellingShingle Lan Zhang
Ulrike M. Stamer
Robin Moolan-Vadackumchery
Frank Stüber
Nuclear Factor-κB Signaling Regulates the Nociceptin Receptor but Not Nociceptin Itself
Cells
cell cultures
cytokines
NFκB
nociceptin
nociceptin receptor
signal transduction
title Nuclear Factor-κB Signaling Regulates the Nociceptin Receptor but Not Nociceptin Itself
title_full Nuclear Factor-κB Signaling Regulates the Nociceptin Receptor but Not Nociceptin Itself
title_fullStr Nuclear Factor-κB Signaling Regulates the Nociceptin Receptor but Not Nociceptin Itself
title_full_unstemmed Nuclear Factor-κB Signaling Regulates the Nociceptin Receptor but Not Nociceptin Itself
title_short Nuclear Factor-κB Signaling Regulates the Nociceptin Receptor but Not Nociceptin Itself
title_sort nuclear factor κb signaling regulates the nociceptin receptor but not nociceptin itself
topic cell cultures
cytokines
NFκB
nociceptin
nociceptin receptor
signal transduction
url https://www.mdpi.com/2073-4409/13/24/2111
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AT robinmoolanvadackumchery nuclearfactorkbsignalingregulatesthenociceptinreceptorbutnotnociceptinitself
AT frankstuber nuclearfactorkbsignalingregulatesthenociceptinreceptorbutnotnociceptinitself