| Objective: To investigate the protective effect and mechanism of Ming Ze Yi Liver Formula(MZYG)on the liver of mice with alcoholic liver disease,and to provide theoretical basis for the clinical treatment of alcoholic liver disease.Methods: Forty Kunming mice were randomly divided into normal group(Normal),model group(Model),Ming Ze Yi Liver low dose group(MZYG-L)and high dose group(MZYG-H),10 mice in each group.Except for the Normal group,all the groups were given52° alcohol for moulding by gavage at regular intervals every day for 4 weeks.Starting from the 5th week,the mice in the Normal and Model groups were given saline and the MZYG-treated group were given Mingze Yihepatic Formula aqueous decoction by gavage for 2 weeks respectively.The liver index was calculated;the levels of ALT,AST,TG,IL-1β,TNF-α,IL-6,ROS,SOD,CAT and GSH-px in the serum of each group were measured by ELISA;the morphological changes of liver tissues were observed by HE staining and oil red O staining;the expression of TLR4 and My D88 were detected by immunohistochemistry.The expression of TLR4,My D88,NF-κB,HMGB1,Bcl-2,bax and P62,Keap1,Nrf2,GPX4,SLC7A11 were detected by Western blot.Results: 1.Liver index: Compared with the Normal group,the liver index of mice in the Model group increased significantly.Compared with the Model group,the liver index of mice in the MZYG-treated group decreased significantly,and the decrease was more obvious in the high-dose group.2.The levels of ALT,AST,TG,IL-1β,TNF-α,IL-6 and ROS in the serum of mice in the Model group were significantly increased and the levels of SOD,CAT and GSH-px were significantly decreased compared with those in the Normal group by ELISA.Compared with the Model group,the serum levels of ALT,AST,TG,IL-1β,TNF-α,IL-6 and ROS were significantly reduced and the levels of SOD,CAT and GSH-px were significantly increased in the MZYG-treated mice.3.Histomorphological changes in the liver: HE staining showed that the hepatocytes in the Model group were disorganized,the texture of the hepatic cords was disorganized,and some of the hepatocytes showed diffuse steatosis,with lipid droplets of different sizes and numbers visible inside,and the liver showed obvious inflammatory cell infiltration;the number of lipid droplets inside the hepatocytes in the MZYG treatment group was significantly reduced,the texture of the hepatic cords was clearly arranged in a radial pattern,and the inflammatory cell infiltration in the liver was significantly reduced.The number of lipid droplets in hepatocytes was significantly reduced in the MZYG-treated group.The hepatocytes in the Model group showed significant fat droplets,while the hepatocytes in the MZYG-treated group showed significantly fewer fat droplets.Immunohistochemical staining,MZYG treatment significantly inhibited the expression of inflammation-related signaling pathway proteins,indicating that,MZYG could significantly reduce the liver damage caused by alcohol.4.Western blot assay: compared with the Normal group,the expression of inflammatory signaling pathway-related proteins TLR4,My D88,p-NF-κBp65,and HMGB1 protein was significantly higher in the liver tissues of mice in the Model group;compared with the Model group,the expression of TLR4,MyD88,p-NF-κBp65,and HMGB1 protein expression was significantly lower in the liver tissues of the MZYG group.This indicated that MZYG could significantly inhibit the alcohol-induced inflammatory response in the liver.Compared with the Normal group,the expression of P62,Nrf2,Bcl-2,SLC7A11,GPX4 and Keap1 and bax proteins were significantly decreased in the liver tissues of the Model group,while the expression of P62,Nrf2,Bcl-2 and Bcl-4 proteins were significantly increased in the liver tissues of the MZYG group.Nrf2,Bcl-2,SLC7A11,GPX4 and Keap1,bax were significantly increased and decreased in the MZYG group,indicating that MZYG could significantly reduce the oxidative stress damage to the liver caused by alcohol.Conclusion: 1.MZYG significantly reduced the levels of AST,ALT,TG and other liver injury-related factors in the serum of mice with alcoholic liver disease,while the degree of hepatocyte steatosis was significantly reduced,indicating that MZYG has a good protective effect on the liver of mice with alcoholic liver disease.2.MZYG significantly reduced the levels of inflammatory factors IL-1β,TNF-α and IL-6 in the serum of mice with alcoholic liver disease,inhibited hepatic inflammatory responses and protected against hepatocyte injury by regulating TLR4/My D88/NF-κBp65related signalling pathways.3.MZYG significantly reduced the serum levels of ROS,increased the levels of antioxidant enzymes SOD,CAT and GSH-Px in mice with alcoholic liver disease,protected against hepatic oxidative stress injury by regulating the P62/Keap1/Nrf2-related signaling pathway,and inhibited hepatocyte apoptosis by suppressing oxidative stress response. |
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