Effect And Mechanism Of Histone Deacetylase 6 Inhibitor ACY-1215 On Nonalcoholic Fatty Liver Disease

Background and objective:Nonalcoholic fatty liver disease(NAFLD)is a chronic liver disease characterized by liver steatosis,which can develop into cirrhosis and hepatocellular carcinoma at the end stage.The number of its patients has increased rapidly in recent years,bringing huge pressure on the social economy and health resources.At present,there is no specific treatment for NAFLD.Changing living habits and diet structure and reducing weight through physical exercise are the mainstream intervention measures.ACY-1215 is a selective inhibitor of histone deacetylation 6,which has shown good therapeutic potential in many tumors,acute liver injury,and alcoholic liver disease.At present,there is no research about ACY-1215 in the treatment of NAFLD.Therefore,this study aims to explore the role and mechanism of ACY-1215 in the experimental model of NAFLD,to propose a new treatment strategy for NAFLD.Methods:(1)Different concentrations of fatty acids were used to co-culture with mouse AML 12 cells and human MIHA cells,the cell growth activity,oil red O staining,and triglyceride(TG)content of cells were used as evaluation indicators to screen the appropriate conditions for establishing the NAFLD cell model.(2)The effect of ACY-1215 on the NAFLD cell model was explored with oil red O staining and cell TG content as evaluation indicators.(3)The AML 12 cell model of NAFLD was detected by TMT relative quantitative proteomics to explore the protein that expression level was changed after the treatment of ACY-1215,and concentrated on the corresponding signal pathway,and preliminarily explore the molecular mechanism of the action of ACY-1215.(4)Lipopolysaccharide was used to stimulate the NAFLD cell model to simulate NAFLD progression,and oil red O staining and cell TG content were used as evaluation indicators;C57BL/6J mice were fed with high fat diet(HFD)to establish an animal model of NAFLD.The body weight,liver samples,HE staining sections,oil red O staining sections,and serum liver enzymes were used as evaluation indicators.To explore the protective effects of ACY-1215 in the NAFLD cell model and animal model.(5)Verify whether ACY-1215 plays a role through the signal pathway explored by TMT relative quantitative proteomics in the cell model and animal model of NAFLD.Results:(1)With the increase of fatty acid concentration and co-culture time,the steatosis produced by hepatocytes would be serious,the content of TG in hepatocytes would rise,and the growing vitality of hepatocytes would decline.The modeling condition of AML 12 cell is co-culture with OA250μmol/L+PA 125μmol/L for 48 hours.The modeling condition of MIHA cell is co-culture with OA 125μmol/L+PA 62.5μmol/L for 48 hours.(2)ACY-1215 can reduce the steatosis produced by the NAFLD cell model and the content of TG in cells,and its effect increases with the increase of ACY-1215 concentration.(3)According to the TMT relative quantitative proteomics analysis,the expression level of 245 proteins in the AML 12 cell model of NAFLD changed under the treatment of ACY-1215,of which 109 proteins were upregulated in the ACY-1215 treatment group and 136 proteins were downregulated in the ACY-1215 treatment group.In the ACY-1215 treatment group,several metabolic pathway signals were changed.It was preliminarily proposed that ACY-1215 inhibited the expression of CD14 and protected NAFLD.(4)ACY-1215 can reduce steatosis,intracellular TG,and the expression level of intracellular inflammatory factor m RNA in the cell model of NAFLD stimulated by LPS.ACY-1215 can reduce the fatty degeneration in the liver,the level of liver enzymes in serum,and the expression level of inflammatory factor m RNA in liver tissue of the HFD-induced NAFLD mice model.(5)ACY-1215 can inhibit CD14/TLR4/My D88/MAPK/NFκB signal pathway-related protein expression levels in the NAFLD cell model and animal model,thereby inhibiting the activation of this signaling pathway.Conclusion:(1)ACY-1215 has protective effects on fatty acid-induced NAFLD cell model and HFD-induced NAFLD animal model.(2)ACY-1215 plays a protective role on NAFLD experimental model by inhibits CD14/TLR4/My D88/MAPK/NFκB signal pathway.