MCC950 improves cell damage and necrosis induced by severe acute pancreatitis by regulating the NLRP3/GSDME signaling pathway
Abulikeli·Wulayin
Maimaitiyiming·Maimaituerxun
Alimu·Maimaiti
Abstract：Objective To explore the molecular mechanism of a novel NOD-like receptor family pyrin domain containing 3(NLRP3)inhibitor(MCC950)in inducing pyroptosis of pancreatic acinar cells(PACs)with an inflammatory response induced by lipopolysaccharide(LPS).Methods Mouse PACs were selected for primary culture in vitro,and monolayer cells in the third generation(P3)were selected for the study.PACs were treated with blank control,LPS,and LPS+low-dose or high-dose MCC950.Fluorescent probes were used to detect reactive oxygen species(ROS)levels.Enzyme-linked immunosorbent assay(ELISA)was performed to measure intracellular inflammatory factors(interleukin-1beta[IL-1β],interleukin-18[IL-18]),and oxidative stress factors(superoxide dismutase[SOD],malondialdehyde[MDA],and glutathione peroxidase[GSH-Px]).RT-PCR and Western blot were used to detect the mRNA and protein expressions of key members in the NLRP3 signaling pathway(NLRP3,apoptosis-associated speck-like protein containing a CARD[ASC],Caspase-1)and pyroptosis-associated markers(Caspase-4,Caspase-5,Caspase-11,gasdermin D[GSDMD],gasdermin E[GSDME]).Results Compared with the control group,LPS induction significantly increased intracellular ROS fluorescence intensity,MDA content,IL-1β and IL-18 levels,and mRNA and protein levels of NLRP3,ASC,Casepase-1,Caspase-4,Caspase-5,Caspase-11,GSDMD and GSDME,and decreased SOD and GSH-Px levels(P<0.05).Compared with LPS-induced PACs,LPS+MCC950 treatment significantly decreased intracellular ROS fluorescence intensity,MDA content,IL-1β and IL-18 levels,and mRNA and protein levels of NLRP3,ASC,Casepase-1,Caspase-4,Caspase-5,Caspase-11,GSDMD and GSDME,and increased SOD and GSH-Px levels(P<0.05).High-dose MCC950 treatment resulted in significantly lower intracellular ROS fluorescence intensity,MDA content,IL-1β and IL-18 levels,and mRNA and protein levels of NLRP3,ASC,Casepase-1,Caspase-4,Caspase-5,Caspase-11,GSDMD and GSDME than the low-dose MCC950 treatment,showing a dose-dependent manner(P<0.05).Conclusion MCC950 can inhibit LPS-induced inflammatory response,oxidative stress injury,and cell pyroptosis in PACs by inhibiting the activity of the NLRP3/GSDME signaling pathway,which deserves further research.
Keywords：acute pancreatitisMCC950cell pyroptosisreactive oxygen species(ROS)NOD-like receptor family pyrin domain containing 3(NLRP3)signaling pathwayoxidative damage
Publication Date：2026-01-26
Online Publishing Date：2026-03-17(First online date of this platform, not the publication date of the document)
Pages：6( 10-14,20 )