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In This Article

  • Summary
  • Abstract
  • Introduction
  • Protocol
  • Representative Results
  • Discussion
  • Acknowledgements
  • Materials
  • References
  • Reprints and Permissions

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 amylase1,2. 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 literatures28,29. Approval for the lack of analgesia was also granted by the Animal Ethics Committee of Soochow University.

1. Preparation

Representative Results

By carefully following the instructions above, we obtained a mean surgery duration of approximately 40 min. The mice were slightly inactive and had lost approximately 0.5-1.75 g, 0.85-1.85 g and 0.5-4.73 g of weight at 24 h, 48 h and 72 h post-operation, respectively (Figure 2).

From the time of surgery completion to 24 h post-operation, as the disease developed, the mice became inactive and showed slow responses and actions.

The surviv.......

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

Acknowledgements

We are grateful for the support from the following grants: a Translational Research Grant of NCRCH [2020WSA01], a KJXW Scientific Grant from Suzhou Commission of Health for Young Scholars [KJXW2020002], a Science and Technology Plan of Suzhou City (SKY2021038 and SKJY2021050), a grant from the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD), and a Primary Research & Social Development Plan of Jiangsu Province (BE2018659).

....

Materials

NameCompanyCatalog NumberComments
0.5% iodophorShanghai Likang Disinfectant3101024 mL/mouse
0.9% sodium chlorideSinopharm Group Co., Ltd.100193180.8 mL/mouse
1% Pentobarbital sodiumSigmaP37610.2 -0.25 mL/mouse
25 μL flat tip Microliter syringeGaoge, ShanghaiA124019
4% ParaformaldehydeBeyotime, Nantong, ChinaP0099-500ml
5% sodium taurocholate (TC)AladdinS100834-5g10 μL/SAP mouse
6-0 Sterile nylon microsuture with threaded needle (1/2 circle)Cheng-He20093
75% alcoholSinopharm Group Co., Ltd.100092184 mL/mouse
8-0 Sterile nylon microsuture with threaded needle (3/8 circle)Cheng-He19064
ALT Activity Assay KitEPNK, Anhui, ChinaALT0012
Amylase Assay KitEPNK, Anhui, ChinaAMY0012
Angled small bulldog clamp with 12 mm jaw (3 cm)Cheng-HeHC-X022
aspen shavings or shreds for mouse beddingBeijing Vital River Laboratory Animal TechnologyVR03015
AST Activity Assay KitEPNK, Anhui, ChinaAST0012
Blood Urea Nitrogen (BUN) Assay KitEPNK, Anhui, ChinaBUN0011
C57BL/6 mouseBeijing Vital River Laboratory Animal Technology213
Creatine Assay KitEPNK, Anhui, ChinaCRE0012
Feature microtome bladeBeyotime, Nantong, ChinaE0994
Hemostatic Forceps (9.5 cm, Curved)JZ, Shanghai Medical Instruments Co. Ltd.JC3901
Lipase Assay KitJiancheng, Nanjing, ChinaA054-2-1
MicrotomeLeica biosystem, GermanyRM2245
Mindray biochemistry analyzerMindray, Shenzhen, ChinaBS-420
MPO Assay KitJiancheng, Nanjing, ChinaA044-1-1
Normal mouse chowTrophic, Nantong, ChinaLAD 1000
Phosphate buffered salineBeyotime, Nantong, ChinaC0221A
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-HeHC-X091A or HC-X090A
Straight Scissors (10.0 cm)Cheng-He, Ningbo, ChinaHC-J039102
Thermo Scientific CentrifugeThermo Scientific, USAMultifuge X1R

References

  1. Lee, P. J., Papachristou, G. I. New insights into acute pancreatitis. Nature Reviews Gastroenterology and Hepatology. 16 (8), 479-496 (2019).
  2. Mandalia, A., Wamsteker, E. J., DiMagno, M. J. Recent advances in understanding and managing acute pancreatit....

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MouseAcute PancreatitisSodium TaurocholateBiliopancreatic DuctRetrograde InjectionAnimal ModelSurgeryAnesthesiaDuodenumPancreasCommon Hepatic DuctMicro injection

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