Establishment of a Mouse Severe Acute Pancreatitis Model using Retrograde Injection of Sodium Taurocholate into the Biliopancreatic Duct

Summary

A mouse model of severe acute pancreatitis is described herein. The procedure presented here is very rapid, simple, and accessible, thereby potentially allowing the study of the molecular mechanisms and different therapeutic interventions in acute pancreatitis in a convenient way.

Abstract

The prevalence of acute pancreatitis (AP), especially severe acute pancreatitis (SAP), is increasing in younger age groups annually. However, there is a lack of effective treatments in the current clinical practice. With the easy accessibility of transgenic and knockout strains and their small size, which allows minimal doses of drugs required for in vivo evaluation, a well-established experimental model in mice is preferred for AP research. Moreover, SAP induced through sodium taurocholate (TC) is currently one of the most widely used and best characterized models. This model has been investigated for novel therapies and possible molecular events during the process of AP. Here, we present the generation of an AP mouse model using sodium taurocholate and a simple homemade microsyringe. Moreover, we also provide the methodology for the subsequent histology and serological testing.

Introduction

Acute pancreatitis (AP) is an acute inflammation of the pancreas characterized by obstruction of the main pancreatic duct with subsequent ductal distension and pancreas autodigestion by its abnormally activated enzymes. Its clinical manifestations include local or systemic inflammation, abdominal pain, and elevation of serum amylase^1,2. According to the severity classification³, AP can present in mild, moderate, and severe forms, and among them, severe acute pancreatitis (SAP) is the most concerning condition due to its high mortality rate of more than 30%⁴. In t....

Protocol

All experiments involving animals were approved by the Animal Ethics Committee of Soochow University. All surgical procedures were performed under full anesthesia. Analgesics were not used to avoid interference with the natural course of the disease according to previous literatures^28,29. Approval for the lack of analgesia was also granted by the Animal Ethics Committee of Soochow University.

1. Preparation

Discussion

The TC-induced SAP model is an excellent research tool. As shown in this study, this model is very easily realized in general labs without employing specific devices. When used in combination with histology and biochemical analysis, it provides a cost- (inexpensive reagents) and time-saving (24 h time window) approach for inducing and evaluating AP. Adjusting the concentration of TC also offers the possibility of producing mild and moderate AP. Perides et al. also employed TC to induce SAP in mice²⁰

Materials

Name	Company	Catalog Number	Comments
0.5% iodophor	Shanghai Likang Disinfectant	310102	4 mL/mouse
0.9% sodium chloride	Sinopharm Group Co., Ltd.	10019318	0.8 mL/mouse
1% Pentobarbital sodium	Sigma	P3761	0.2 -0.25 mL/mouse
25 μL flat tip Microliter syringe	Gaoge, Shanghai	A124019
4% Paraformaldehyde	Beyotime, Nantong, China	P0099-500ml
5% sodium taurocholate (TC)	Aladdin	S100834-5g	10 μL/SAP mouse
6-0 Sterile nylon microsuture with threaded needle (1/2 circle)	Cheng-He	20093
75% alcohol	Sinopharm Group Co., Ltd.	10009218	4 mL/mouse
8-0 Sterile nylon microsuture with threaded needle (3/8 circle)	Cheng-He	19064
ALT Activity Assay Kit	EPNK, Anhui, China	ALT0012
Amylase Assay Kit	EPNK, Anhui, China	AMY0012
Angled small bulldog clamp with 12 mm jaw (3 cm)	Cheng-He	HC-X022
aspen shavings or shreds for mouse bedding	Beijing Vital River Laboratory Animal Technology	VR03015
AST Activity Assay Kit	EPNK, Anhui, China	AST0012
Blood Urea Nitrogen (BUN) Assay Kit	EPNK, Anhui, China	BUN0011
C57BL/6 mouse	Beijing Vital River Laboratory Animal Technology	213
Creatine Assay Kit	EPNK, Anhui, China	CRE0012
Feature microtome blade	Beyotime, Nantong, China	E0994
Hemostatic Forceps (9.5 cm, Curved)	JZ, Shanghai Medical Instruments Co. Ltd.	JC3901
Lipase Assay Kit	Jiancheng, Nanjing, China	A054-2-1
Microtome	Leica biosystem, Germany	RM2245
Mindray biochemistry analyzer	Mindray, Shenzhen, China	BS-420
MPO Assay Kit	Jiancheng, Nanjing, China	A044-1-1
Normal mouse chow	Trophic, Nantong, China	LAD 1000
Phosphate buffered saline	Beyotime, Nantong, China	C0221A
Straight micro-bulldog clamp with 5 mm jaw (1.5 cm)	JZ, Shanghai Medical Instruments Co. Ltd.	W40130
Straight or curved forceps (11.0 cm)	Cheng-He	HC-X091A or HC-X090A
Straight Scissors (10.0 cm)	Cheng-He, Ningbo, China	HC-J039102
Thermo Scientific Centrifuge	Thermo Scientific, USA	Multifuge X1R