Induction of acute pancreatitis model by Ceruletide

caerulein（Ceruletide）Animal disease model induced acute pancreatitis

Introduction:

acute pancreatitis(Acute pancreatitis, AP)It is a common acute abdomen in clinical practice, with complex causes and a mortality rate of up to5%~10%Severe illnessAPThe mortality rate is as high as20%~30%At present, the pathogenesis has not been elucidated and there is no specific treatment method. Currently liveAPThe commonly used methods for animal model production include pancreatic duct ligation, duodenal loop closure, pancreatic duct puncture injection, pancreatic submucosal injection, pancreatic duct injection combined with intravenous infusion, and intraperitoneal injection of frog skin extract and lipopolysaccharide.

caerulein(Caerulein)It is a substance similar in function and composition to cholecystokinin(cholecystokinin，CCK)Gastric regulatory molecules stimulate excessive secretion of pancreatic acinar cells, leading to obstacles in the separation of trypsinogen and lysosomal hydrolases within the cells. Through the use of tissue proteasesBActivation occurs in a dependent manner, and the activity of pancreatic enzymes can further lead to the activation of zymogens, causing a series of cell damage caused by protease activity. The inflammatory response caused by self digestion of pancreatic tissue recruits inflammatory cells, releases inflammatory factors, and further causes severe local or even systemic inflammatory reactions.

Operation process:

1. select6Zhou Ling's masculinityC57BL/6Mice, raised inSPFLevel environment, maintain12Hour light dark cycle, temperature 20℃Humidity40%-60%Free to eat and drink.

2. Implementation of mold making

1）Animal pretreatment

Before modeling8Fasting treatment (without water) was performed on mice at an hour to reduce the interference of gastrointestinal contents on pancreatic detection.

2）Preparation of rain frog extract

Dilute rain frog extract with physiological saline to the working concentration.

3）Intraperitoneal injection

press100ug/kgDose of weight, per1Inject intraperitoneally into mice at an hour1Secondary rain frog hormone, continuous injection8Multiple low-dose injections can simulate the progressive onset of acute pancreatitis in humans, avoiding acute mortality caused by a single high-dose injection.

4)blank control

The control group mice were intraperitoneally injected with an equal amount of physiological saline at the same time to eliminate the stress effects of the injection procedure itself.

5）Observation and sample collection after modeling

After modeling, closely observe the mental state, activity level, food and water intake, and fecal characteristics of the mice to evaluate their overall stress response. Injecting rain frog hormone24After hours, euthanize the mice and collect key samples:

a.pancreatic tissue

Quickly separate the pancreas, weigh and partially fix it in place10%Neutral formalin (used for pathological sections), partially frozen in liquid nitrogen (used for molecular detection).

b.blood sample

After collecting blood from the eye socket, centrifuge and separate the serum for detecting amylase, lipase, and inflammatory factors.

c.Other samples

According to research needs, samples such as colon and feces can be collected for correlation analysis.

3.Model validation:

1)Pathological injury of pancreatic tissue

a.throughHEStaining was used to observe the morphological changes of the pancreas, and the model group showed structural disorder, edema, and necrosis of pancreatic acinar cells, as well as infiltration of a large number of inflammatory cells.

b.Model group pancreas/An increase in body weight indicates pancreatic swelling and injury.

2)Elevated levels of serum digestive enzymes

The serum amylase and lipase activities in the model group were significantly higher than those in the blank control group, and the difference was statistically significant（p<0.0001）Confirmed that damage to pancreatic acinar cells leads to the release of digestive enzymes into the bloodstream.

3)Release of serum pro-inflammatory cytokines

Model group serum pro-inflammatory factorsIL-6TheTNF-The significant increase in alpha concentration indicates that rain frog extract successfully induced systemic spread of pancreatitis.

Product Information

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