Effects Of Ursodeoxycholic Acid On Liver Inflammatory Lesions And Blood Glucose In Rats With Bile Duct Ligation

Objective:To investigate the effects of ursodeoxycholic acid on liver inflammatory lesions and blood glucose in rats with bile duct ligation.Methods:Thirty-two healthy male SD rats aged 6-8 weeks were randomly divided into four groups: group A(normal group,n = 8),group B(sham operation group,n = 8)without ligation of bile duct after laparotomy,group C(experimental group,n = 8)underwent bile duct ligation after laparotomy,group D(intervention group,n = 8)on the basis of group C,ursodeoxycholic acid intervention(0.25% of food intake)was given 5 days after operation.Once a day).The general state of SD rats in each group was observed.Group A,B,C and D were weighed at 4 weeks after operation,liver weight was weighed during treatment,liver index(liver weight / body weight,%)was calculated,glucose tolerance test was measured,serum total bile acid(TBA),aspertate aminotransferase(AST),alanine aminotransferase(ALT)and albumin(ALB)were measured.Liver HE staining and masson staining were used to observe the occurrence of liver fibrosis.The expression of hepatic inflammatory cytokines NF-KB-p65 and IKB α in each group was detected by immunohistochemistry.Results:Three days after operation,there was no significant change in daily food intake,sclera,urine color and general mental state between group B and group A.obvious jaundice was found in eyes,ear tip and tail in group C compared with group B.the color of urine deepened and became dark yellow,the daily food intake decreased and the stool color became lighter in group D.there was no significant difference between group D and group C.Four weeks after operation,there was no significant change in daily food intake,sclera,urine color and general mental state between group B and group A.The spirit of group C was slightly worse and the daily food intake was less than that of group B,yellow staining could still be seen on the edge of the ear,the color of urine was roughly the same,and the color of stool was slightly lighter and drier.Four weeks later,the rats in group C opened the abdominal cavity and could see the filled bile duct;compared with group C,the spirit of group D was similar,the degree of yellow staining of ear margin was lighter,the daily food intake was increased,and the color of urine and stool were basically the same.Compared with group A,there was no significant difference in the weight gain rate of group B(4 weeks after operation-0 weeks after operation)/ 0 weeks after operation(P > 0.05).Compared with group B,the weight gain rate of group C was significantly lower than that of group B(P < 0.05),and that of group D was significantly higher than that of group C(P < 0.05).There was no significant difference in liver index between group B and group A(P > 0.05).The liver index in group C was significantly higher than that in group B(P < 0.05),and the liver index in group D was lower than that in group C(P > 0.05).Group A,B,C and D were all treated after 4 weeks.The blood was collected from the heart,and the supernatant was centrifuged to determine the indexes of ALT,AST,TBA and ALB.Compared with group A,ALT,AST,TBA and ALB in group B had no significant changes(P>0.05);AST,ALT and TBA in group C were significantly higher than those in group B after 4 weeks(P<0.05),and ALB in group C was higher than that in group B Significantly decreased(P<0.05);ALB in group D was significantly higher than that in group C(P<0.05),and AST and ALT in group D tended to decrease compared with group C(P>0.05).HE staining under microscope showed: in group B,the hepatic lobules were neatly arranged in cords,the bile ducts were not significantly expanded,and the liver cells were not obviously necrotic.In group C,bile ducts were dilated,fibrous cords were seen around them,and the structure of hepatic lobules was disordered.In group D,the bile ducts were slightly dilated,and there were no obvious fibrous cords around them.Masson staining stains collagen fibers blue.Masson staining observation under microscope: liver lobules in group B were neatly arranged,the cytoplasm was stained dark red,and there was no obvious blue around.In group C,there were obvious blue-stained fibrous cords around the bile ducts.In group D,slight staining was seen in the hepatic lobules and around the bile ducts.The liver tissue wax blocks were selected,and the immunohistochemistry of liver NF-KB-p65 and IKB α antibodies could be used to characterize and quantify the local inflammation in SD rat liver.The results are as follows: the expression of NF-KB-p65 antibody in group C under the microscope was deeper than that in group B,and the local nuclei were more deeply stained and the positive area was larger.Compared with group C,NF-KB-p65 antibody in group D showed local nuclei.Compared with group B,the expression of IKB α antibody in group C did not change significantly.Compared with group C,the IKB α antibody in group D had deeper staining and larger positive area.We use Image J software,which can calculate the positive expression area(Area)and the total optical density value(IOD value),and finally we can obtain the average optical density value(AOD value)by calculating IOD/Area.By comparing the AOD values of each group,we can clarify the expression quantity of NF-k Bp65 antibody and IKB α antibody in each group,and calculate the average optical density value of NF-KB-P65(AOD1 value)and the average optical density value of IKB α(The ratio of AOD2 values)can compare the inflammatory response among the three groups.According to the analysis of statistical results,the ratio of AOD1/AOD2 in group C was significantly increased compared with that in group B(P<0.05),and the ratio of AOD1/AOD2 in group D was significantly lower than that in group C(P<0.05).Four weeks after the operation,the blood glucose of group B at 0h,1h,and 2h had no significant change compared with that of group A(P>0.05),and the blood glucose of group C at 0h,1h,and 2h had a tendency to increase compared with that of group B(P>0.05).,the 1h and 2h blood glucose in group D were significantly lower than those in group C(P<0.05),and the 0h blood glucose in group D had no significant change compared with group C(P>0.05).The area under the OGTT curve(AUC)was calculated from the OGTT chart of SD rats in each group.According to the results,the AUC of group B had no significant change compared with group A(P>0.05),and the AUC of group C was significantly higher than that of group B(P<0.05).),the AUC of group D was significantly lower than that of group C(P<0.05).Conclusion:Through bile duct ligation in rats,the NF-KB inflammatory pathway is activated,and liver fibrosis changes;blood sugar increases,and insulin resistance occurs.After 4weeks of ursodeoxycholic acid intervention,it was found that the expression of NF-KB-p65 was decreased,and liver fibrosis was improved;blood glucose was decreased,and insulin resistance was improved.