Study On Glucose Metabolism And Islet Function Changes In Mice With Liver Cirrhosis

Objective:Liver is an important organ of glucose metabolism,and many factors can affect the metabolic of glucose,impair the glucose tolerance,and even the appearance of diabetes.In1906,Narmyn defined this type of diabetes to chronic liver parenchyma as hepatogenous diabetes(HD).According to statistics,about 50%-80%of patients with chronic liver disease have abnormal glucose tolerance,and about 30%of them eventually develop diabetes,while the incidence of diabetes in normal population is only about 8%.However,the pathogenesis of HD is not clear,and its clinical manifestation is affected by the original liver disease,which is easily ignored,but the specific mechanism is unclear.In this paper,a model of liver cirrhosis was established through the ligation of mice choledochosis,to explore the changes of glucose metabolism and islet function in cirrhosis,the change of key enzyme activity of sugar metabolism and the role of inflammatory factors.Methods:Select the size of a male 6~8 weeks of age 60 c57BL/6 mices,randomly divided into 6 groups,the one experimental group(group A1,n=10),the other experimental group(group A1,n=10),the one sham-operation group(group B1,n=10),the other sham-operation group(group B2,n=10),the one normal group(group C1,n=10),the other normal group(group C2,n=10).In the aseptic conditions,the mice in the A1 and A2groups were opened with bile duct ligation,and the mice in group B1 and B2 opened the free bile duct in the abdominal cavity but did not ligate,and the mice in the C1 and C2groups did not open the stomach and did not deal with the bile duct.Each group of mice measured fasting blood glucose weekly and weighed it,and looked at the general condition and color of urine and feces.Mices in group A1,B1,C1 and in group A2,B2,C2 were separately weighted,under anesthesia condition,glucose tolerance test and insulin release experiment were determined for evaluating islet function,serum aspertate aminotransferase(AST),alanine aminotransferase(ALT),TG(triglyceride),albumin concentrations were determined by microplate methods,HE and masson staining were used to observe whether these mice develop liver cirrhosis,the activity of glucose kinase(GCK)and glucose 6phosphatase(G6PC)in liver tissue homogenate were measured with ELISA method,inflammatory factors NF-KB-P65,I-KB?in the liver and pancreas were determined using immunohistochemical method in 4~thh and 8~thh weeks after the bile duct ligation.Results:Experiment in the 4~thh weeks,1)compared with C1 group and B1 group tip of ear and the tail of mice in experimental group were visible yellow dye,urine were dark yellow,stools were light color,fur slightly lose luster,activity decreased,weight were reduced obviously(P<0.05);2)Serum AST and ALTwere elevated(P<0.05),there wasno obvious changes in TG,albumin,liver weight,OGTT area under the curve(P>0.05),the area under the insulin release curve was increased(P<0.05).Liver HE and masson staining demonstrated small bile duct dilation,with a small amount of fibrosis around,and partial hepatic lobules with broken ring,indicating necrotic necrosis of liver cells;3)the activity of GCK and the activity of G-6-PC were not changed in liver tissue homogenate(P>0.05);4)Immunohistochemical demonstrated phos-NF-KB expression in the liver and pancreatic tissue were increased.the number of pancreatic islets in the pancreas was reduced and the area was reduced,but there was no obvious changes(P>0.05),and immunohistochemical demonstrated IKB?expression in the liver and pancreatic tissue were increased and the nucleus were stained the brown.Experiment in the 8~thh weeks,1)compared with C2 group and B2 group whole body yellowing obviously,dark yellow urine,stool color becomes shallow,fur more lacklustre,weight were reduced obviously(P<0.01);2)Serum AST and ALTwere elevated obviously(P<0.01),liver weight was elevated(P<0.05),albuminwas reduced obviously(P<0.01),there was no obvious changes in TG and OGTT area under the curve(P>0.05),the area under the insulin release curve was increased(P<0.05).Liver HE and masson staining the bile duct was obviously dilated,and the surrounding fibrosis was obvious,and part of it extended to the leaves of the liver to form false lobules,which showed large necrosis of liver cells.3)The activity of G-6-PC was elevated in liver tissue homogenate(P<0.05),the activity of GCK was not changed.4)The number of pancreatic islets in the pancreas was decreased and the area was reduced,and the liver and pancreatic tissue phos-NF-KBandIKB?wereincreasedinbrownhyperchromaticcells,phos-NF-KB/IKB?was elevated(P<0.05).Conclusion:we built mice mode of liver cirrhosis successfuly bybile duct ligation,mice with liver cirrhosis develoed insulin resistance after the 4~thh week.With the progress of the liver cirrhosis,the activity of G-6-PC was elevated,activatedthe NF-KB signaling pathways,inflammation occurred in liver and pancreas.The number of pancreatic islets in the pancreas was decreased and the area was reduce.