Specific inhibition of NLRP3 inflammasome by a Smurf1 inhibitor in vitro and in vivo

Objectives: Abnormal activation of the NLRP3 inflammasome is associated with various inflammatory diseases, making it a potential therapeutic target. A01 is an inhibitor of Smurf1, its effect on the activation of NLRP3 inflammasome is still unclear and needs further study.

Material and methods: Caspase-1 and IL-1β were detected by immunoblotting and ELISA, and (Lactate Dehydrogenase) LDH release was measured via a specific kit. Immunoprecipitation was used to explore the interaction between NLRP3 and ASC. In a mouse model of alum-induced peritonitis, PECs were collected and analyzed by flow cytometry. In the high-fat diet (HFD) model, blood glucose was measured with a glucometer.

Results: A01 inhibited the activation of the NLRP3 inflammasome in macrophages without affecting the activation of AIM2 or NLRC4 inflammasomes. Mechanistically, A01 suppressed NLRP3 inflammasome assembly and activation by disrupting the NLRP3-ASC interaction. Moreover, A01 demonstrated protective effects in mouse models of NLRP3 inflammasome-mediated diseases.

Conclusions: A01 specifically suppresses NLRP3 inflammasome activation in vitro and in vivo. A01 disrupts the association of NLRP3 and ASC. These findings suggest that A01 is a specific NLRP3 Inhibitor and may be a promising candidate for treating NLRP3 inflammasome-related diseases.