Effects And Mechanisms Of Anti Hepatic Fibrosis Of Ethanolic Extracts Of Euonymus Alatus On Mice Induced By CCl₄ Based On The AMPK/mTOR Signaling Pathway

Objective:To study the effects and mechanisms of Ethanolic Extracts of Euonymus Alatus（EEEA）on Hepatatic fibrosis（HF）model mice induced by CCl₄.Methods:50 C57BL/6J male mice were randomly divided into 5 groups,10 in each group,namely,normal group（NG）,model group（MG）,low,medium and high-dose group（EEEA-L,EEEA-M,EEEA-H）.In addition to NG,the other four groups were molded with CCl₄mixed liquid.At the same time of molding,NG and MG filled the stomach with double steam water,and the remaining three groups were filled with the stomach at the corresponding dose:EEEA-L,EEEA-M and EEA-H were 2g/kg,4g/kg,and 8g/kg respectively.30 days later,the mouse liver index was calculated,the serum and liver of mice were collected,the liver function indexes in the serum were detected by AST and ALT test boxes,the content of TNF-αand IL-6 in the serum were detected by ELISA method,the liver damage and collagen deposition in mice were detected by HE and Masson staining,the relative expression of protein in AMPK,m TOR,α-SMA and Collagen I in liver tissue by Western blot method,and the relative expression of AMPK,m TOR,α-SMA and Collagen I in liver tissue was detected by RT-q PCR method.Results:（1）Liver function results show that compared with NG,the serum content of AST and ALT in MG mice increased significantly（P<0.05）,and compared with MG,the serum content of AST and ALT in the three groups of EEEA decreased significantly（P<0.05）.（2）ELISA results show that compared with NG,the serum content of TNF-αand IL-6 in MG mice increased significantly（P<0.05）,compared with MG,the serum content of TNF-αin the three groups of EEEA decreased significantly（P<0.05）,and the content of IL-6 in the serum of EEEA-M and EEEA-H decreased significantly（P<0.05）.（3）The liver index results show that compared with NG,the MG liver index is significantly higher（P<0.05）,and compared with MG,the EEEA-H liver index decreases significantly（P<0.05）.（4）HE staining results show that compared with NG,MG mice hepatocyte structure is destroyed,the hepatic cord arrangement is disordered,the hepatic lobulary structure is destroyed,and more inflammatory cells can be seen between tissues.Compared with MG,the three groups of EEEA improved the hepatocyte structure,hepatic cord arrangement,hepatic lobulary structure and inflammatory cell infiltration in mice to varying degrees.（5）Masson staining results show that compared with NG,MG hepatocytes have obvious edema deformation,and more collagen fiber deposition is seen in the confluence area.Compared with MG,the three groups of EEEA improved mouse hepatocytes and collagen deposition to varying degrees.（6）Western Blot results show that compared with NG,the relative expression ofα-SMA,Collagen I and m TOR proteins of MG increased significantly（P<0.05）.Compared with MG,the three groups of EEA reduced the relative expression ofα-SMA,Collagen I and m TOR proteins（P<0.05）to varying degrees.Compared with NG,the relative expression of AMPK protein in MG decreased significantly（P<0.05）,and compared with MG,the relative expression of AMK was increased by the three groups of AMK to varying degrees（P<0.05）.（7）RT-q PCR results show that compared with NG,the expression of AMPK m RNA in MG decreased significantly（P<0.05）,and the expression of AMPK m RNA in the three groups of EEEA increased significantly（P<0.05）;compared with NG,the expressions ofα-SMA,Collagen I and m TOR m RNA in MG increased significantly（P<0.05）,and the expressions ofα-SMA,Collagen I and m TOR m RNA in the three groups of EEEA decreased significantly decreased significantly（P<0.05）.Conclusion:EEEA can improve liver injury and liver fibrosis in mice induced by CCl₄.Its mechanism may be related to the expression of proteins and m RNA that activate AMPK and inhibit m TOR.