Protocol for in vivo and in vitro activation of NLRP3 inflammasome in mice using monosodium urate

STAR Protoc. 2023 Sep 15;4(3):102554. doi: 10.1016/j.xpro.2023.102554. Epub 2023 Sep 7.

Abstract

Although intraperitoneal injection of monosodium urate (MSU) is an effective model for studying peritonitis, its establishment remains challenging. Here, we present a protocol for using MSU to activate the NLRP3 inflammasome in bone-marrow-derived macrophages (BMDMs) and to induce peritonitis in mice. We describe steps for isolating and culturing BMDMs, preparing MSU crystals, and activating the NLRP3 inflammasome using western blot and ELISA. We then detail procedures for inducing peritonitis and testing for relevant indicators using flow cytometry and ELISA. For complete details on the use and execution of this protocol, please refer to Huang et al. (2023).1.

Keywords: Cell Biology; Immunology; Metabolism.

MeSH terms

  • Animals
  • Inflammasomes*
  • Macrophages
  • Mice
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • Peritonitis* / chemically induced
  • Uric Acid / pharmacology

Substances

  • Inflammasomes
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • Uric Acid