| Objective:KM mice with severe obstructive jaundice were used as experimental research models to observe the intervention effect of silybin on the liver injury caused by obstructive jaundice in mice,and to explore the mechanism of its effect on the differential expression of factors related to the Bcl-2/Bax-Caspase-3 signaling pathway by affecting the expression of miR-497.Methods:Thirty male KM mice were randomly divided into model group?M?,silybin treatment group?SFJB?,and sham operation group?SO?,with 10 mice in each group.Group M and SFJB constructed the mouse model of severe obstructive jaundice liver injury by using a new common bile duct suspension method of our research group.The operation method of the SO group was the same as that of group M,but the common bile duct was not suspended.After the obstruction of common bile duct was relieved 48 hours after the operation,the SFJB group was given 200mg·kg-1·d-1 silibin by gavage,and the other two groups were given normal saline with equal volume for 7 weeks.In the last time after drug intervention,the night the process groups of mice were banned from food,drink,be sodium pentobarbital anesthesia,the second day morning to remove eye take blood,in groups of mice serum aspertate alanine transaminase?ALT?,aspartate transaminase?AST?,total bilirubin?TBIL?,total bile acid?TBA?,alkaline phosphatase?ALP?levels;The complete liver was taken out,and the liver tissue samples were embedded and sectioned for HE staining.The pathological changes of the liver tissues in each group were observed and compared under the microscope.The differentially expressed proteins of Bcl-2,Bax and Caspase-3 in liver tissues were detected by immunohistochemistry.The expression levels of Bcl-2,Bax and Caspase-3 in liver tissue were determined by ELISA.The expression levels of Bcl-2,Bax,Caspase-3mRNA and miR-497 in liver tissues were detected by qPCR.Results:?1?General conditions:The general conditions of mice in the SO group were normal;Mice in group M showed scratch marks in their hair,lethargy,decreased activity,yellow staining in the tips of their ears,tail and sclera,yellow urine,and lighter stools.The situation of SFJB group was better than that of M group.?2?Serology:Compared with the SO group,the serum levels of ALT,AST,TBIL,TBA and ALP in the M group were increased?P<0.01?,while those in the SFJB group were decreased?P<0.01?.?3?HE staining:The cords of hepatocytes and sinusoids in the SO group were arranged neatly around the central vein,and there were a few red blood cells and inflammatory cells in the sinusoids.In group M,there was spot-like necrosis of hepatocytes,a large number of nuclear pyknotic cells,hepatic cord structure disorder,and a large number of red blood cells and inflammatory cells in sinusoids.In the SFJB group,the liver cells were poorly arranged,the inflammatory cell infiltration was reduced,and no necrotic cells were observed.?4?The immunohistochemical staining results:The positive expression areas of Bax and Caspase-3 proteins in the liver tissues of mice in the SFJB group and SO group were lower than those in the M group?P<0.01?,and the positive expression areas of Bcl-2 were higher than those in the M group?P<0.01?.?5?ELISA results:Compared with group M,Bcl-2 protein content in group SO and SFJB increased?P<0.01?,Bax and Caspase-3protein content decreased?P<0.01?.?6?qPCR results:Compared with group M,Bcl-2 mRNA expression in group SO and SFJB increased?P<0.01?,Bax and Caspase-3 mRNA expression and miR-497 content decreased?P<0.01?.Conclusion:Silybin has better improvement effect on liver injury caused by obstructive jaundice in mice and can obviously reduce the degree of liver injury.The mechanism of its anti-injury action may be related to the differential expression of related factors in the Bcl-2/Bax-Caspase-3 signaling pathway regulated by miR-497. |
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