Amitriptyline inhibits NLRP3 inflammasome activation via the ASM/CE pathway in a cell model of NAFLD  

作　　者：QIN LIU CHUNYAN NIU QIANG ZHANG SHIQIN SUN YUE CHEN YONGQIANG SHI 

机构地区：[1]Shengli Clinical Medical College,Fujian Medical University,Fuzhou,350001,China [2]Department of Gastroenterology,Nanjing Lishui People’s Hospital,Zhongda Hospital Lishui Branch,Southeast University,Nanjing,211200,China [3]Department of Gastroenterology,Yancheng Third People’s Hospital(The Yancheng School of Clinical Medicine of Nanjing Medical University),Yancheng,224000,China [4]Administrative Department of Teaching and Scientific Research,Nanjing Lishui People’s Hospital,Zhongda Hospital Lishui Branch,Southeast University,Nanjing,211200,China

出　　处：《BIOCELL》2024年第5期759-769,共11页生物细胞（英文）

基　　金：supported by the Initial Scientific Research Fund of the Talents Introduced in Nanjing Lishui People’s Hospital(Project 2021YJ02).

摘　　要：Background:Nonalcoholic fatty liver disease(NAFLD)is a global health concern with the acid sphingomyelinase(ASM)/ceramide(CE)pathway and the NOD-like receptor family,pyrin domain-containing protein 3(NLRP3)inflammasome identified as pivotal players in lipid disorders and inflammation.This study explores the interaction mechanism between the ASM/CE pathway and NLRP3 in NAFLD cell models,aiming to understand the impact of amitriptyline(Ami),an ASM inhibitor,on lipid deposition and hepatocyte injury by regulating the ASM/CE-NLRP3 pathway.Methods:HepG2 and HL-7702 cells were exposed to free fatty acids(FFAs)to establish the NAFLD model.The cells were divided into 5 groups:control group,model group,Ami group,tumor necrosis factoralpha(TNF-α)group,and Ami+TNF-αgroup.Intracellular lipid droplets were visualized using Oil Red O staining,and Western blot analysis quantified ASM,NLRP3,and caspase 1 protein expression.Enzyme linked immunosorbent assay(ELISA)was measured CE and ASM levels,while qRT-PCR assessed mRNA expression.The apoptotic rate was evaluated by flow cytometry(FCM).Results:Following FFAs incubation,significant increases in ASM and CE levels were observed in HepG2 and HL-7702 cells,accompanied by elevated expression of NLRP3,and caspase 1,and IL-1β.TNF-αtreatment further amplified these indicators.Ami demonstrated a reduction in lipid deposition,suppressed ASM/CE pathway activation,downregulated NLRP3 and caspase 1 expression,and improved apoptosis.Additionally,MCC950,a selective inhibitor of the NLRP3,mitigated NLRP3,caspase 1,and IL-1βexpression,alleviating lipid deposition and apoptosis in the NAFLD cell model.Conclusion:The ASM/CE-NLRP3 pathway in NAFLD cells promotes hepatocyte steatosis,inflammation,and cell damage.Ami emerges as a promising therapeutic agent by inhibiting the ASM/CE-NLRP3 pathway,underscoring its potential as a key target for NAFLD treatment.

关 键 词：Nonalcoholic fatty liver disease HEPATOCYTE AMITRIPTYLINE ASM/CE pathway NLRP3 Nonalcoholic steatohepatitis 

分 类 号：R575.5[医药卫生—消化系统]