Wuxi School of Medicine, Jiangnan University, Jiangsu 214000, China.
Wuxi School of Medicine, Jiangnan University, Jiangsu 214000, China.
STAR Protoc. 2023 Sep 15;4(3):102554. doi: 10.1016/j.xpro.2023.102554. Epub 2023 Sep 7.
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)..
虽然腹腔内注射单钠尿酸盐 (MSU) 是研究腹膜炎的有效模型,但它的建立仍然具有挑战性。在这里,我们提出了一个使用 MSU 激活骨髓来源的巨噬细胞 (BMDM) 中的 NLRP3 炎性体并在小鼠中诱导腹膜炎的方案。我们描述了分离和培养 BMDM、制备 MSU 晶体以及使用 Western blot 和 ELISA 激活 NLRP3 炎性体的步骤。然后,我们详细介绍了诱导腹膜炎和使用流式细胞术和 ELISA 检测相关指标的程序。有关该方案使用和执行的完整详细信息,请参阅 Huang 等人。(2023)。