How to prepare experimental animal models of acute pancreatitis?
动物模型,急性胰腺炎
z-bo
2020-07-29
623
The animal model of pancreatitis is the best way to study the etiology of pancreatitis and explore clinical treatment methods. However, since the cause of pancreatitis varies with the type of pancreatitis, the cause of pancreatitis is still unknown, so there is no good method. The animal model can simulate human pancreatitis. However, Busnar-do et al. successfully induced an acute pancreatitis model in 1856 by injecting a mixture of bile and olive oil into the pancreatic duct of a dog. It mainly includes the following contents: 1. Sodium taurocholate modeling
1) The method of establishing the model: The rats are fasted for 12 hours before the operation, and are anesthetized with free drinking water and 2 mg/100 g body weight of pentobarbital solution And fixed. Enter the abdomen through a midline incision in the upper abdomen, lift the duodenum, and fix the duodenal segment and the hilar pancreaticobiliary duct with small blood vessel clips. Puncture the pancreaticobiliary duct with a TB needle (4.5 needle, 1 ml syringe) (preferably using a micro pump injection), inject 5% C sodium taurocholate (0.1 ml/100 g) for 1 minute, let it stand for 5 minutes, and then pull it out Take out the needle and remove the vascular clip. Observe the edema and hemorrhage of rat pancreatic tissue, and suture all layers of the abdominal wall.
2) Model features and functions: This method is a more mature model of acute necrotizing pancreatitis (ANP).
3) Precautions for model establishment and use: The 24-hour survival rate is about 50%
2. Frogpisu model
1) How to build a model: Fast the rats for 12 hours before the operation and drink free water. , Frog skin (25μg/kg) intraperitoneal injection, 5 hours after injection, the formation of edematous acute pancreatitis (CIP)
2) Model features and functions: This method of mild edematous acute pancreatitis is a model.
3) Model establishment and usage instructions: At present, ANP is mainly conducting research in China. Please pay attention to your research when using this model.
3. Retrograde bile duct injection
The pancreatic duct is ligated, inserted into the main pancreatic duct, and injected through the main pancreatic duct. Sodium deoxycholate, autologous bile and other substances that can activate pancreatin or pancreatin. Acute necrotizing pancreatitis can occur when PGE2 is injected at the same time. Schmid et al. established a rat model of acute pancreatitis using small-dose pancreatic perfusion of glycodeoxycholic acid combined with intravenous injection of bombesin. This may result in uniform damage to the gland cells, accompanied by moderate necrosis, suitable for the study of pancreatitis.
Four. Pancreaticobiliary duct ligation
The pancreatic duct is ligated to the duodenum, or only the pancreatic duct is ligated. Lerch and colleagues ligated the pancreatic duct of the ampulla. Edema pancreatitis appeared 6 hours after ligation, bleeding, necrosis, and inflammatory cells appeared 12 hours after infiltration. The advantage of this model is that it is simple and avoids the influence of drugs on non-specific systems. It is similar to human bile reflux pancreatitis, but only mild pancreatitis can occur. Unzi et al. improved this method. After cutting the proximal end of the duodenum 10 cm, the two ends were closed and the common bile duct was ligated. When the stomach and duodenum are anastomosed, pancreatic juice flows into the closed loop, and when the internal pressure increases, reflux occurs. Acute necrotizing pancreatitis develops, edematous pancreatitis occurs 4 hours after surgery, and hemorrhage and necrosis occurs 9-12 hours after surgery. Acute pancreatitis also exists due to pancreatic duct ligation or obstruction of pancreatic arteries or veins.
5. Feeding method
Choline ethionine is not sensitive to the stimulation of female and male rats and can cause acute pancreatitis model. The young mice received choline-free ethionine and pancreatic necrosis, and abdominal fat necrosis occurred 4 days later. The basic principle is that the choline-free ethionine can block the secretion of pancreatic acinar, zymogen granules and lysosomes are activated in the cell.
6. Methods to promote secretion
Bombesin analog CCK-PZ via intravenous or subcutaneous injection. It can significantly increase the secretion of pancreatic proteolytic enzymes to a level that causes pancreatic acinar autolysis.
7. Isolated pancreatitis model
mainly use isolated extracorporeal perfusion technology, free pancreas, duodenum and spleen. Insert the artery, mesenteric artery, portal vein and pancreatic duct, remove the pancreas, and connect to an oxygen and perfusion circulator that controls temperature and humidity. After 30 minutes of stable perfusion, free fatty acids were injected from the artery to simulate alcoholic pancreatitis and block the pancreatic duct and vein. Bombesin injection can cause cholangiopancreatitis and ischemic pancreatitis for 2 hours.
