Naloxone inhibits nod-like receptor protein 3 inflammasome

Han Yu Lin, Ya Ying Chang, Ming Chang Kao, Chun Jen Huang

Research output: Contribution to journalArticle

Abstract

Background Naloxone, an opioid receptor antagonist, possesses potent anti-inflammation effects. We previously confirmed the effects of naloxone on inhibiting upregulation of inflammatory cytokine interleukin-1β (IL-1β). Production of mature form IL-1β is mediated by the nod-like receptor protein 3 (NLRP3) inflammasome, a multiprotein complex composed of NLRP3, and the adaptor protein apoptosis-associated speck-like protein contains a caspase recruitment domain (ASC). We elucidated whether naloxone could inhibit the activation of NLRP3 inflammasome. Material and methods To induce IL-1β production and NLRP3 inflammasome activation, the human monocytic leukemia cell line THP-1 cells were first primed with lipopolysaccharide (LPS, 1 μg/mL) and then activated with adenosine triphosphate (ATP, 1 mM). For NLRP3 transcription, THP-1 cells were only treated with LPS priming. Results Enzyme-link immunosorbent assay data revealed that the concentration of IL-1β in THP-1 cells treated with LPS plus ATP was significantly higher than that in THP-1 cells treated with LPS plus ATP plus naloxone (0.1 μM) (P < 0.001). Real-time quantitative reverse transcription and polymerase chain reaction data also revealed that NLRP3 mRNA concentration in THP-1 cells treated with LPS was significantly higher than that in THP-1 cells treated with LPS plus naloxone (P = 0.001). ASC speck formation, that is, ASC assembles into a large protein complex, is an indicator for NLRP3 inflammasome activation. Our data revealed that the percentage of cells containing ASC specks in THP-1 cells treated with LPS plus ATP was also significantly higher than that in THP-1 cells treated with LPS plus ATP plus naloxone (P < 0.001). Conclusions Naloxone inhibits NLRP3 inflammasome activation.

Original languageEnglish
Pages (from-to)72-77
Number of pages6
JournalJournal of Surgical Research
Volume219
DOIs
Publication statusPublished - Nov 1 2017
Externally publishedYes

Fingerprint

Inflammasomes
Naloxone
Adenosine Triphosphate
Proteins
Interleukin-1
Multiprotein Complexes
Immunosorbents
Narcotic Antagonists
Reverse Transcription
Lipopolysaccharides
Leukemia
Up-Regulation

Keywords

  • ASC
  • IL-1β
  • Inflammasome
  • Naloxone
  • NLRP3
  • THP-1 cell

ASJC Scopus subject areas

  • Surgery

Cite this

Naloxone inhibits nod-like receptor protein 3 inflammasome. / Lin, Han Yu; Chang, Ya Ying; Kao, Ming Chang; Huang, Chun Jen.

In: Journal of Surgical Research, Vol. 219, 01.11.2017, p. 72-77.

Research output: Contribution to journalArticle

Lin, Han Yu ; Chang, Ya Ying ; Kao, Ming Chang ; Huang, Chun Jen. / Naloxone inhibits nod-like receptor protein 3 inflammasome. In: Journal of Surgical Research. 2017 ; Vol. 219. pp. 72-77.
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AB - Background Naloxone, an opioid receptor antagonist, possesses potent anti-inflammation effects. We previously confirmed the effects of naloxone on inhibiting upregulation of inflammatory cytokine interleukin-1β (IL-1β). Production of mature form IL-1β is mediated by the nod-like receptor protein 3 (NLRP3) inflammasome, a multiprotein complex composed of NLRP3, and the adaptor protein apoptosis-associated speck-like protein contains a caspase recruitment domain (ASC). We elucidated whether naloxone could inhibit the activation of NLRP3 inflammasome. Material and methods To induce IL-1β production and NLRP3 inflammasome activation, the human monocytic leukemia cell line THP-1 cells were first primed with lipopolysaccharide (LPS, 1 μg/mL) and then activated with adenosine triphosphate (ATP, 1 mM). For NLRP3 transcription, THP-1 cells were only treated with LPS priming. Results Enzyme-link immunosorbent assay data revealed that the concentration of IL-1β in THP-1 cells treated with LPS plus ATP was significantly higher than that in THP-1 cells treated with LPS plus ATP plus naloxone (0.1 μM) (P < 0.001). Real-time quantitative reverse transcription and polymerase chain reaction data also revealed that NLRP3 mRNA concentration in THP-1 cells treated with LPS was significantly higher than that in THP-1 cells treated with LPS plus naloxone (P = 0.001). ASC speck formation, that is, ASC assembles into a large protein complex, is an indicator for NLRP3 inflammasome activation. Our data revealed that the percentage of cells containing ASC specks in THP-1 cells treated with LPS plus ATP was also significantly higher than that in THP-1 cells treated with LPS plus ATP plus naloxone (P < 0.001). Conclusions Naloxone inhibits NLRP3 inflammasome activation.

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