Background Intrinsic immunity is an important line of defense against pathogenic microorganisms , which can effectively fight and remove external dangers. However, dysfunction of innate and adaptive immunity is considered to be a key step in the initiation and maintenance of autoimmune diseases. NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome is a multimeric protein complex, which can detect exogenous pathogen irritants and endogenous danger signals, which promote interleukin (IL)-1β and IL-18 secretion and pyroptosis mediated by caspase-1. Numerous studies have shown their significance in autoimmune diseases, such as rheumatoid arthritis (RA) and inflammatory bowel disease (IBD), which strongly indicate that NLRP3 inflammasome complex may serve as a promising and novel therapeutic target for clinical treatment in inflammatory-related diseases.

Methods THP-1, PBMC and monocyte-induced macrophages are the main model cells in cellular experiments in vitro. For THP-1, which is usually induced into macrophage-like cells before being tested; primary macrophage can be Macrophages can be induced by PBMC-isolated monocytes. LPS is generally used as the primary source of stimulation to activate cells and cause NLRP3 priming, and compounds can be used to inhibit NLRP3 prior to the use of Nigericin or ATP to stimulate cellular secretion of IL-1β or IL-18, and compound activity is assessed by assaying the amount of cytokine secreted.

Results Whether it is THP-1 cells, PBMC or monocyte-induced macrophages, once NLRP3 is inhibited after LPS stimulation, it can effectively inhibit the secretion of IL-1β or IL-18, as well as caspase-1 activity, and by analyzing the amount of cytokine secretion or caspase1 activity, we can assess the inhibitory activity of different types of compounds on NLRP3.

Conclusions Based on the function of NLRP3, we have established different methods for evaluating the activity of various NLRP3 inhibitors in different cell systems.