Guizhou Miniature Pig Model Of Biliary Liver Fibrosis

Objective: To establish Guizhou miniature pigs cholestatic liverfibrosis model by bile duct ligation; observation the changes of liver functionand histopathology in the model animals,discuss the expression and mechanismof Fas protein in the liver cells of animal models. Methods: Eight femaleweighing12to15kg of Guizhou miniarure pigs,3%pentobarbital (1ml/kg)intraperitoneal injection of anesthesia, laparotomy revealed the common bileduct and free, double common bile duct ligation and transection of the commonbile duct between the two lines, we have established an animal model.30minutes before surgery and postoperative7days，14days and28days to takeblood samples for testing liver function,routine blood and coagulation; beforebile duct ligation, postoperative7days,14days and28days，we take livertissue for Optical microscopic and electron microscopy examination;immunohistochemical detection of Fas expression in specimens of each period;Tunel situ method to detect cell apoptosis. Results: Pig cholestatic liver fibrosismodel successfully established by this method. Two pigs died due to liverfailure at1week and3weeks, the remaining six survived to cut the abdominalaorta were sacrificed at four weeks, survival pigs are different degrees of liverfibrosis. Within28days after ligation, compared with the preoperative serummarkers aspartate aminotransferase (AST) in the7days after that to a highervalue (394.2±124.8U/L), compared with preoperative (139.8±66.9U/L)were significantly different (P <0.01), and then showed a gradually increasing trend;alanine aminotransferase (ALT) after7days increased (65.0±1.6U/L),compared with preoperative (46.35±1.4U/L) were significantly different (P<0.01);28days after surgery, the serum total protein and albumin slow decline,compared with preoperative significant difference (P <0.01); Total bilirubin(TBIL) and direct the bilirubin (DBIL) content continues to rise from7daysafter surgery (111.4±11.0umol/L and93.2±8.2umol/L),compared withpreoperative (3.1±1.7umol,/L and1.4±0.6umol/L) were significantlydifferent (P <0.01); Alkaline phosphatase (ALP) from7days after the surgerycontinues to rise (599.3±64.0U/L), compared with the preoperative(161.5±21.4U/L) were significantly different (P <0.01);White blood cells(WBC) from7days after the surgery continues to rise (30.0±5.7×10E9/L), compared withthe preoperative(11.7±1.8×10E9/L) were significantly different (P <0.01),28days after surgery, Red blood cells(RBC)and Hemoglobin(HGB) slow decline,compared with preoperative significant difference (P <0.05); Prothrombin time(PT), thrombin time (TT) and fibrinogen (dFbg) slow decline in the28daysafter surgery, compared with preoperative significant difference (P <0.01);28days after surgury, the optical microscope examinations showed gradualdestruction of lobular architecture, interlobular fibrous hyperplasia,disorganized, the electron microscopic examination showed a large number oflipid droplets in liver cells, the rough endoplasmic reticulum disappears theperinuclear gap widened, the nuclear membrane shrinkage, mitochondrialedema; Pathological examination and immunohistochemistry:7days after bile duct ligation, liver cell necrosis as the main performance, over time, thehepatocyte apoptosis and expression of Fas increased, severity of of liverfibrosis increased; Conclusion: We can establish pigs cholestatic liver fibrosismodel by the common bile duct ligation; its mechanisms related to theexpression of FAS increased and promote liver cell apoptosis.