Molecular Mechanism Of Morin On Alleviating Non-alcoholic Fatty Liver Disease In Mice

Non-alcoholic fatty liver disease(NAFLD)is the most common chronic liver disease,which is considered to be the concentrated expression of metabolic disorders in the liver.NAFLD includes simple fatty liver or non-alcoholic fatty liver(NAFL)and non-alcoholic fatty liver disease(NASH).It is characterized by abnormal deposition of large amounts of fat and is often associated with metabolic diseases such as obesity and type 2 diabetes mellitus.Serious NAFLD can develop into liver cirrhosis and liver cancer.Oleic acid(OA),a kind of free fatty acids is one of the main substances causing NAFLD,which can induce lipid accumulation in human hepatocellular carcinoma(Hep G2)cells.Surfactant Tyloxapol can inhibit the activity of lipase and is often used as a stimulant to establish hyperlipidemia model in mice.Therefore,this study assessed the potential therapeutic effect of MO on NAFLD and its molecular mechanism by using the inflammation and lipid accumulation of Hep G2 cells induced by OA and Tyloxapol-induced lipid elevation and hepatic steatosis in mice.In vitro,MTT assay was used to screen the safe concentration of drugs,oil red O staining was used to determine the degree of intracellular lipid accumulation,and the contents of TNF-α and triglyceride(TG)were measured.Western Blot was used to detect the expression of related proteins and determine the effect of MO on cell model and molecular mechanism.The results showed that OA-induced lipid accumulation in Hep G2 cells was significant,TG level and TNF-α release were significantly increased,but MO could significantly inhibit these changes;Western Blot results showed that MO significantly increased PPARα protein expression,inhibited SREBP-1c protein expression,and increased the phosphorylation levels of ACC,AKT and AMPK;MO also inhibited MAPKs/NF-κB inflammation signaling pathway;In vivo experiments,Tyloxapol significantly increased plasma triglyceride(TG)and total cholesterol(TC)levels in mice,and resulted in a large number of lipid droplets accumulated in mouse hepatocytes.MO significantly inhibited the increase of plasma TC and TG levels induced by Tyloxapol,and alleviated balloon-like changes and lipid accumulation in hepatocytes.Western Blot experiment showed that MO up-regulated PPARα protein expression and inhibited it.The protein expression of SREBP-1c was produced and phosphorylated expression of ACC,AKT and AMPK was activated.In conclusion,MO can play a significant role in anti-lipid accumulation,and it is expected to play a role in the mechanism of NAFLD and other metabolic diseases and the development of candidate drugs.