Effects Of Silybin On MiRNA-122 Mediated Wnt Pathway On Liver Fibrosis In Obstructive Jaundice Mice

Background and purpose:Obstructive jaundice is mainly caused by pancreaticobiliary duct disease.If left untreated,it will cause persistent cholestasis.Patients’ serum bilirubin and bile acid levels will increase,which induces intracellular inflammation and hepatic fibrosis.Tissues have abnormal hyperplasia,which leads to liver fibrosis and even cirrhosis.In recent years,domestic and foreign studies on the Wnt signaling pathway and liver disease have become hot spots.A large number of research results have confirmed that the abnormal activation of the Wnt/β-catenin signaling pathway is involved in liver inflammation,cirrhosis,liver fibrosis,and benign and malignant liver The occurrence and development of diseases such as tumors.As an important microRNA that is highly expressed in the liver,miRNA-122 has been proved to be an effective biomarker for liver disease,which can target the expression of Wnt1 gene.Silybin is a kind of medicine for protecting liver and protecting liver.It has the functions of regulating lipid metabolism,anti-oxidation,scavenging free radicals,and stabilizing cell membrane.However,its specific mechanism in the process of repairing liver fibrosis damage caused by jaundice is still worthy of deepening discussion.Based on the above conclusions,this experiment was to observe the effect of silybin on hepatic fibrosis in obstructive jaundice mice and the expression of Wnt1,β-catenin and GSK-3β in liver tissues in the classic Wnt signaling pathway mediated by miRNA-122 Difference,to explore the mechanism of silybin on liver fibrosis caused by obstructive jaundice.Methods:Thirty male KM mice were randomly divided into a model group(M),a silibinin group(SFJB),and a sham operation group(SO),with 10 in each group.Except for the sham operation group,the other groups were made of obstructive jaundice using the common bile duct suspension method,and the obstruction was relieved after 48 hours.The SFJB group was administered with a 20 mg/ml silybin solution at a dose of 0.2 mg/g/d Stomach,the other two groups of mice were given an equal volume of saline.After 7 weeks of intervention,blood was removed by eyeball removal and liver tissue was isolated.The contents of aspartate aminotransferase(AST),alanine aminotransferase(ALT),total bilirubin(TBIL),total bile acid(TBA)and alkaline phosphatase(ALP)in serum of each group of mice were determined by spectrophotometry.Detection;HE staining to observe pathological changes in liver tissue;immunohistochemical staining to detect differential expression of Wnt1,β-catenin,GSK-3β in liver tissue;enzyme-linked immunosorbent assay(ELISA)to detect Wnt1,β-catenin,GSK-3β protein content;Realtime-PCR detected Wnt1,β-catenin,GSK-3β,p-GSK3β,miRNA-122 levels in liver tissue.Results:(1)48 hours after the operation,the mice in the model group and silibinin group had poor appetite,and the hair,mouse tail,and ear tips were severely yellow-stained,and the urine was dark yellow;the mice in the sham-operated group had a normal diet,but no hair,mouse tail,or ear tips.Yellow staining,pale yellow urine.(2)Compared with the model group,the AST,ALT,TBIL,TBA and ALP levels in serum of the silibinin group decreased(P<0.01).(3)The results of HE staining showed that the hepatocytes in the sham operation group were arranged radially,and the hepatic cord and sinus structure were clear;in the model group,hepatocytes in the model group were widened,disordered,hepatic cords were broken,hepatocytes showed nuclear contraction,and tissue gaps were seen.Inflammatory cell infiltration and red blood cell accumulation,and many spindle-shaped myofibroblasts were visible;in the silibinin group,hepatic lobular structures were irregular,and hepatic cords were arranged irregularly.The spindle-shaped myofibroblasts were reduced compared to the model group,and liver lobular The structural change was significantly improved compared with the model group.(4)The results of immunohistochemistry showed that Wnt1,β-catenin and GSK-3β had the lowest expression in the sham operation group,less positive staining,and the distribution range was scattered;the highest expression in the model group,more positive staining,and a wider distribution range;and compared with the model group,the expression levels of all factors in the silybin group decreased significantly,and the area of positive staining decreased significantly(P<0.01).(5)ELISA test results showed that the Wnt1,β-catenin,and GSK-3β protein contents were the lowest in the sham operation group and the highest in the model group.Compared with the model group,the Wnt1,β-catenin,and GSK-3β protein contents in the silybin group were significant decrease(P<0.01).(6)Realtime-PCR test results showed that compared with the sham operation group,Wnt1,β-catenin,and GSK-3β mRNA levels in the liver of the model group were significantly increased(P<0.01),and the silybin group was lower than the model group(P<0.01);The miR-122 mRNA content was the lowest in the sham operation group.Compared with the sham operation group,the miR-122 mRNA content in the model group was significantly higher(P<0.01),and the silybin group was higher than the model group(P <0.01).Conclusion:Silibinin can improve the liver fibrosis of mice with obstructive jaundice.The mechanism may be related to silibinin up regulating the expression of mirna-122 and down regulating the expression of Wnt1,β-Catenin and GSK-3 β in Wnt / β-Catenin pathway.