Effects of N-acetylcysteine on hepatic inflammatory response and pyroptosis in rats with alcoholic liver disease based on NLRP3 inflammasome
10.13431/j.cnki.immunol.j.20240112
- VernacularTitle:基于NLRP3炎性小体探究N-乙酰半胱氨酸对酒精性肝病大鼠肝脏炎性反应及焦亡的作用
- Author:
Xiaojing ZHU
1
;
Hailin TANG
;
Liang ZHOU
Author Information
1. 215101,苏州市中西医结合医院药剂科
- Publication Type:Journal Article
- Keywords:
Alcoholic liver disease;
N-acetylcysteine;
Pyroptosis;
NOD-like receptor pyrin domain-containing protein 3;
Reactive oxygen species
- From:
Immunological Journal
2024;40(11):803-811
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effects of N-acetylcysteine(NAC)on liver tissue inflammation,pyroptosis levels,and NOD-like receptor pyrin domain-containing protein 3(NLRP3)inflammasome in rats with alcoholic liver disease(ALD).Methods Male SD rats were divided into a normal group,an ALD model group,a 100 mg·kg-1 NAC group,and a 200 mg·kg-1 NAC group.HE staining was used to observe pathological changes in liver tissue,and TUNEL method was used to detect the apoptosis rate of liver cells.Biochemical kits were used to measure serum levels of aspartate aminotransferase(AST),alanine aminotransferase(ALT),and triglyceride(TG).Normal rat hepatocytes BRL-3A were divided into a control group,a 200 mmol/L alcohol group(model group),a 2 mmol/L NAC group,and a 5 mmol/L NAC group.MTT method was used to detect the survival rate of BRL-3A cells;Western blot was used to detect the protein expression levels of cleaved caspase-1,apoptosis-associated speck-like protein containing a CARD(ASC),N-terminal gasdermin D(GSDMD-N),and NLRP3.Total superoxide dismutase(SOD)activity detection kit and malondialdehyde(MDA)kit were used to measure SOD and MDA levels.ELISA was used to detect IL-1β and IL-18 levels.Fluorometric assay were used to measure reactive oxygen species(ROS)level in liver tissue,and immunofluorescence staining was used to determine ROS level in BRL-3A cells.Results Compared with the normal group,the model group showed diffuse fatty degeneration in the liver,increased serum levels of ALT,AST and TG,elevated levels of IL-1β and IL-18 in liver tissue,increased ROS and MDA levels,and upregulated protein expression levels of cleaved caspase-1,ASC,GSDMD-N and NLRP3,while a decrease in SOD level.Compared with the model group,the NAC groups exhibited reduced fatty degeneration in the liver,decreased expression levels of pyroptosis-related proteins,reduced levels of IL-1β and IL-18,lower ROS and MDA levels,and increased SOD level.Compared with the control group,the model group of BRL-3A cells showed increased levels of IL-1β and IL-18 in the supernatant,elevated ROS and MDA levels in cells,and upregulated pyroptosis-related protein expression,while a decrease in SOD level.These indicators were reversed in the NAC group as compared to the model group.Conclusion NAC suppresses liver tissue inflammation and pyroptosis in ALD rats,possibly through blocking the NLRP3 inflammasome signaling pathway.
