Study On The Mechanism Of Wedelolactone Improving Alcohol-induced Liver Injury In Mice

Objective:To investigate the protective effect and mechanism of Wedelolactone on alcoholic liver injury in mice.Method:According to the Gao-binge model,which is a mouse model of alcoholic liver injury using chronic alcohol feeding plus acute alcohol gavage,C57BL/6J male mice of 8-10 weeks were selected for weighing,and the mice were randomly divided into 6 groups according to body weight,5-8 mice in each group,including the control group(Control,control liquid feed),the model group(Ethanol,alcohol liquid feed),the low-dose Wedelolactone group(WED11,The dosage Wedelolactone was 11 mg/kg,control liquid feed),the high-dose Wedelolactone group(WED22,The dosage Wedelolactone was 22 mg/kg,control liquid feed),the low-dose Wedelolactone alcohol group(E+W11,The dosage Wedelolactone was 11 mg/kg,alcohol liquid feed),and the high-dose Wedelolactone alcohol group(E+W22,The dosage Wedelolactone was 22mg/kg,alcohol liquid feed).First,each group was given a LieberDeCarli control diet(35%fat calories)for 5 days.On day6,Ethanol,E+W11 and E+W22 were replaced with Lieber-DeCarli 5%(v/v)alcohol liquid feed for 10 days.During the 10 days,mice in the WED 11 group and the E+W11 group were given daily gavage of 11mg/kg of Wedelolactone,mice in the WED22 group and the E+W22 group were given daily gavage of 22mg/kg of Wedelolactone,and mice in the Control group and the Ethanol group were given daily gavage the same amount of 0.5%of tween-80 solution.On the 15th day.Ethanol,E+W1l and E+W22 were given 5g/kg Ethanol by gavage,while Control,WED 11 and WED22 were given equal heat and volume dextrin.After 9 hours,the mice were killed and samples of blood,liver and fat were collected.HE staining was used to observe the liver pathological changes in mice.The activity of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in serum and the content of triglyceride(TG)and total cholesterol(TC)in the liver were determined by enzymatic method.The serum adiponectin(ADPN)content was measured by ELISA.The malondialdehyde(MDA)content in liver tissue was measured by colorimetric method.Using real-time fluorescence quantitative PCR(qRT-PCR)to detect the expression of related genes.Western blot was used to test the expression of liver tissue related proteins.Result:1.The final weight of mice in the Ethanol group and E+W11 group and E+W22 group decreased significantly from the initial weight.Compared with the Control group,HE staining results showed that hepatocytes in the Ethanol group had increased and increased big fat vacuoles.Hepatic cell steatosis was significantly increased,and hepatocytes were hyalinized.Mallory bodies were visible in the cells.Infiltration of inflammatory cells in the liver lobes.In the E+W1 1 and E+W22 group,the hepatocytes of the mice were significantly steatosis,with mixed steatosis as the main component;some hepatocytes had increased eosinophils and had vitreous changes.2.Compared with the Control group,the serum ALT and AST enzyme activities of the Ethanol group mice increased,the content of ADPN increased,and the contents of TG,TC and MD A in the liver tissue increased.Compared with the Ethanol group mice,in the E+W22 group,the serum ALT and AST enzyme activity decreased,ADPN content decreased,liver TC and MDA content decreased.3.Compared with the Control group,the Ethanol group of liver tissues interleukin 1β(IL1β)and interleukin 6(IL-6)and monocyte chemokine 1(MCP-1)and macrophage inflammatory protein 1α(MIP-1α)and macrophage inflammatory protein 1β(MIP-1β)were significantly increased.Compared with the Ethanol group,IL-1β,IL-6,MCP-1,MIP-1α,MIP-1β and NF-κB p65 phosphorylation levels were significantly down-regulated in the E+W22 group.4.Compared with the Control group,the expression of glutathione peroxidase 4(GPx4)in the Ethanol group was decreased.Compared with the Ethanol group,the expression of hepatic GPx4 was increased in the E+W22 group.5.Compared with the Control group,the adiponectin receptor 2(AdipoR2)expression in the liver was decreased,the adiponectin expressed in the adipose was decreased in the Ethanol group.Compared with the Ethanol group,the expression of hepatic adiponectin receptor 1(AdipoR1),and AdipoR2 were all increased,the adiponectin expressed in the adipose was increased in the E+W22 group.6.Compared with the Control group,the NF-κB p65 phosphorylation level of the Ethanol group was increased.Compared with the Ethanol group,the NF-κB p65 phosphorylation level of the E+W22 group was decreased.Conclusion:Wedelolactone can improve liver inflammation in alcoholic liver injury mice,and its mechanism may be related to the regulation of GPX4 and adiponectin signaling pathways,and the inhibition of NF-κB inflammatory signaling pathways.