Glucose regulates hypoxia‐induced NLRP3 inflammasome activation in macrophages

• Published in: Journal of cellular physiology 2020-10, Vol.235 (10), p.7554-7566
• Main Authors: Watanabe, Sachiko, Usui‐Kawanishi, Fumitake, Karasawa, Tadayoshi, Kimura, Hiroaki, Kamata, Ryo, Komada, Takanori, Inoue, Yoshiyuki, Mise, Nathan, Kasahara, Tadashi, Takahashi, Masafumi
• Format: Article
• Language: English
• Subjects: Adenosine; Adenosine triphosphate; Adenosine Triphosphate - metabolism; Animals; Apoptosis; Apoptosis - drug effects; Atomic absorption analysis; Atomic absorption spectroscopy; Caspase; Caspase 1 - metabolism; Cell activation; Cell death; Cell Death - drug effects; Cells, Cultured; Deprivation; Efflux; Glucose; Glucose - metabolism; Glycolysis; Hypoxia; Hypoxia - metabolism; Inflammasomes; Inflammasomes - metabolism; Inflammation; Inflammation - metabolism; Inflammatory diseases; Interleukin-1beta - metabolism; Interleukins; interleukin‐1; Intracellular; L-Lactate dehydrogenase; Lactate dehydrogenase; Lactic acid; Lipopolysaccharides; Lipopolysaccharides - pharmacology; Macrophages; Macrophages - drug effects; Macrophages - metabolism; Male; Mice; Mice, Inbred C57BL; NLR Family, Pyrin Domain-Containing 3 Protein - metabolism; Nucleotides; Oligomerization; Pathogenesis; Potassium; Potassium - metabolism; potassium efflux; Pyrin protein; Pyroptosis; Regulatory mechanisms (biology); Signal Transduction - drug effects; Signal Transduction - physiology; Spectral analysis; Spectrometry
• ISSN: 0021-9541; 1097-4652
• DOI: 10.1002/jcp.29659

Although the intimate linkage between hypoxia and inflammation is well known, the mechanism underlying this linkage has not been fully understood. Nucleotide‐binding oligomerization domain‐like receptor (NLR) family pyrin domain containing 3 (NLRP3) inflammasome is an intracellular multiprotein complex that regulates interleukin‐1β (IL‐1β) secretion and pyroptosis, and is implicated in the pathogenesis of sterile inflammatory diseases. Here, we investigated the regulatory mechanism of NLRP3 inflammasome activation in response to hypoxia in macrophages. Severe hypoxia (0.1% O2) induced the processing of pro‐IL‐1β, pro‐caspase‐1, and gasdermin D, as well as the release of IL‐1β and lactate dehydrogenase in lipopolysaccharide (LPS)‐primed murine macrophages, indicating that hypoxia induces NLRP3 inflammasome‐driven inflammation and pyroptosis. NLRP3 deficiency and a specific caspase‐1 blockade inhibited hypoxia‐induced IL‐1β release. Hypoxia‐induced IL‐1β release and cell death were augmented under glucose deprivation, and an addition of glucose in the media negatively regulated hypoxia‐induced IL‐1β release. Under hypoxia and glucose deprivation, hypoxia‐induced glycolysis was not driven and subsequently, the intracellular adenosine triphosphates (ATPs) were depleted. Atomic absorption spectrometry analysis showed a reduction of intracellular K+ concentrations, indicating the K+ efflux occurring under hypoxia and glucose deprivation. Furthermore, hypoxia and glucose deprivation‐induced IL‐1β release was significantly prevented by inhibition of K+ efflux and KATP channel blockers. In vivo experiments further revealed that IL‐1β production was increased in LPS‐primed mice exposed to hypoxia (9.5% O2), which was prevented by a deficiency of NLRP3, an apoptosis‐associated speck‐like protein containing a caspase recruitment domain, and caspase‐1. Our results demonstrate that NLRP3 inflammasome can sense intracellular energy crisis as a danger signal induced by hypoxia and glucose deprivation, and provide new insights into the mechanism underlying hypoxia‐induced inflammation. Hypoxia induces nucleotide‐binding oligomerization domain‐like receptor (NLR) family pyrin domain containing 3 (NLRP3) inflammasome activation, resulting in subsequent interleukin‐1β release and pyroptotic cell death in macrophages. Hypoxia‐induced NLRP3 inflammasome activation is markedly influenced by glucose levels.