NLRP3 Inflammasome-Mediated Neutrophil Extracellular Trap Release Regulates Cardiomyocyte Apoptosis in Septic Mice
Description
The cecal ligation and puncture (CLP) method was adopted to establish a sepsis model in C57BL/6 mice. Wild-type (WT) and NLRP3 homozygous knockout (NLRP3 KO) mice were randomly divided into the sham operation group, CLP model group, and CLP + MCC950 intervention group. The survival rate of mice was recorded continuously for 7 days. Small-animal ultrasound was used to detect cardiac function indicators. Hematoxylin-eosin (HE) staining was performed to observe the pathological damage of myocardial tissue, and Wright-Giemsa staining was used to assess the formation of NETs in neutrophils. Western blot was applied to detect the expression levels of NLRP3 pathway-related proteins in neutrophils and apoptosis-related proteins in myocardium. ELISA was used to determine the levels of inflammatory factors, NETs biomarkers and myocardial injury markers in serum and myocardial tissue. In vitro experiments, primary neutrophils were treated with LPS combined with MCC950 to verify the regulatory effect of the NLRP3 pathway on NETs release. Additionally, cardiomyocytes were stimulated with exogenous NETs at different concentrations to detect changes in cell viability and apoptosis level.