Name: establishment of a mouse severe acute pancreatitis model using retrograde injection of sodium taurocholate into the biliopancreatic duct
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Description: Watch this Scientific Journal Video about Establishment of a Mouse Severe Acute Pancreatitis Model using Retrograde Injection of Sodium Taurocholate into the Biliopancreatic Duct at JoVE.com

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 &#38; Social Development Plan of Jiangsu Province (BE2018659).

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
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<ol>
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A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Classification+of+acute+pancreatitis-2012:+revision+of+the+Atlanta+classification+and+definitions+by+international+consensus.">Classification of acute pancreatitis-2012: revision of the Atlanta classification and definitions by international consensus.</a> Gut. 62 (1), 102-111 (2013).</li><li>Munir, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Advances+in+immunomodulatory+therapy+for+severe+acute+pancreatitis.">Advances in immunomodulatory therapy for severe acute pancreatitis.</a> Immunology Letters. 217, 72-76 (2020).</li><li>Peery, A. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Burden+of+gastrointestinal+disease+in+the+United+States:+2012+update.">Burden of gastrointestinal disease in the United States: 2012 update.</a> Gastroenterology. 143 (5), 1179-1187 (2012).</li><li>Hines, O. J., Pandol, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Management+of+severe+acute+pancreatitis.">Management of severe acute pancreatitis.</a> BMJ. 367, 6227 (2019).</li><li>James, T. W., Crockett, S. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Management+of+acute+pancreatitis+in+the+first+72+hours.">Management of acute pancreatitis in the first 72 hours.</a> Current Opinion in Gastroenterology. 34 (5), 330-335 (2018).</li><li>Silva-Vaz, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Murine+models+of+acute+pancreatitis:+a+critical+appraisal+of+clinical+relevance.">Murine models of acute pancreatitis: a critical appraisal of clinical relevance.</a> International Journal of Molecular Sciences. 20 (11), 2794 (2019).</li><li>Hyun, J. J., Lee, H. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+models+of+pancreatitis.">Experimental models of pancreatitis.</a> Clinical Endoscopy. 47 (3), 212-216 (2014).</li><li>Renner, I. G., Wisner, J. R., Rinderknecht, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protective+effects+of+exogenous+secretin+on+ceruletide-induced+acute+pancreatitis+in+the+rat.">Protective effects of exogenous secretin on ceruletide-induced acute pancreatitis in the rat.</a> Journal of Clinical Investigation. 72 (3), 1081-1092 (1983).</li><li>Niederau, C., Ferrell, L. D., Grendell, J. 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M., Werner, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+alcohol-induced+pancreatic+injury+is+similar+with+intravenous+and+intragastric+routes+of+alcohol+administration.">Acute alcohol-induced pancreatic injury is similar with intravenous and intragastric routes of alcohol administration.</a> Pancreas. 43 (1), 69-74 (2014).</li><li>Huang, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fatty+acid+ethyl+ester+synthase+inhibition+ameliorates+ethanol-induced+Ca2+-dependent+mitochondrial+dysfunction+and+acute+pancreatitis.">Fatty acid ethyl ester synthase inhibition ameliorates ethanol-induced Ca2+-dependent mitochondrial dysfunction and acute pancreatitis.</a> Gut. 63 (8), 1313-1324 (2014).</li><li>Sun, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NRF2+mitigates+acute+alcohol-induced+hepatic+and+pancreatic+injury+in+mice.">NRF2 mitigates acute alcohol-induced hepatic and pancreatic injury in mice.</a> Food and Chemical Toxicology. 121, 495-503 (2018).</li><li>Kui, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+insights+into+the+methodology+of+L-arginine-induced+acute+pancreatitis.">New insights into the methodology of L-arginine-induced acute pancreatitis.</a> PLoS One. 10 (2), 0117588 (2015).</li><li>Creutzfeldt, W., Schmidt, H., Horbach, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Studies+on+the+effects+of+a+trypsin+inhibitor+(Trasylol)+on+Enzyme+activities+and+morphology+in+taurocholate+and+calciphylaxis+pancreatitis+of+the+rat+(a+contribution+to+the+pathogenesis+of+pancreatitis).">Studies on the effects of a trypsin inhibitor (Trasylol) on Enzyme activities and morphology in taurocholate and calciphylaxis pancreatitis of the rat (a contribution to the pathogenesis of pancreatitis).</a> Klin Wochenschr. 43, 15-22 (1965).</li><li>Liu, Z. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+taurocholate-induced+model+of+severe+acute+pancreatitis+in+rats.">A simple taurocholate-induced model of severe acute pancreatitis in rats.</a> World Journal of Gastroenterology. 15 (45), 5732-5739 (2009).</li><li>Cavdar, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controversial+issues+in+biliary+pancreatitis:+when+should+we+perform+MRCP+and+ERCP.">Controversial issues in biliary pancreatitis: when should we perform MRCP and ERCP.</a> Pancreatology. 14 (5), 411-414 (2014).</li><li>Perides, G., van Acker, G. J., Laukkarinen, J. M., Steer, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+acute+biliary+pancreatitis+induced+by+retrograde+infusion+of+bile+acids+into+the+mouse+pancreatic+duct.">Experimental acute biliary pancreatitis induced by retrograde infusion of bile acids into the mouse pancreatic duct.</a> Nature Protocols. 5 (2), 335-341 (2010).</li><li>Orabi, A. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cluster+of+differentiation+38+(CD38)+mediates+bile+acid-induced+acinar+cell+injury+and+pancreatitis+through+cyclic+ADP-ribose+and+intracellular+calcium+release.">Cluster of differentiation 38 (CD38) mediates bile acid-induced acinar cell injury and pancreatitis through cyclic ADP-ribose and intracellular calcium release.</a> Journal of Biological Chemistry. 288 (38), 27128-27137 (2013).</li><li>Fanczal, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=TRPM2-mediated+extracellular+Ca(2+)+entry+promotes+acinar+cell+necrosis+in+biliary+acute+pancreatitis.">TRPM2-mediated extracellular Ca(2+) entry promotes acinar cell necrosis in biliary acute pancreatitis.</a> Journal of Physiology. 598 (6), 1253-1270 (2020).</li><li>Huang, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Caffeine+protects+against+experimental+acute+pancreatitis+by+inhibition+of+inositol+1,4,5-trisphosphate+receptor-mediated+Ca2++release.">Caffeine protects against experimental acute pancreatitis by inhibition of inositol 1,4,5-trisphosphate receptor-mediated Ca2+ release.</a> Gut. 66 (2), 301-313 (2017).</li><li>Zhang, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dehydrocholic+acid+ameliorates+sodium+taurocholate-induced+acute+biliary+pancreatitis+in+mice.">Dehydrocholic acid ameliorates sodium taurocholate-induced acute biliary pancreatitis in mice.</a> Biology and Pharmaceutical Bulletin. 43 (6), 985-993 (2020).</li><li>Hagiwara, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antithrombin+III+prevents+cerulein-induced+acute+pancreatitis+in+rats.">Antithrombin III prevents cerulein-induced acute pancreatitis in rats.</a> Pancreas. 38 (7), 746-751 (2009).</li><li>Hagiwara, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Danaparoid+sodium+prevents+cerulein-induced+acute+pancreatitis+in+rats.">Danaparoid sodium prevents cerulein-induced acute pancreatitis in rats.</a> Shock. 32 (1), 94-99 (2009).</li><li>Wittel, U. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Taurocholate-induced+pancreatitis:+a+model+of+severe+necrotizing+pancreatitis+in+mice.">Taurocholate-induced pancreatitis: a model of severe necrotizing pancreatitis in mice.</a> Pancreas. 36 (2), 9-21 (2008).</li><li>Barlass, U., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Morphine+worsens+the+severity+and+prevents+pancreatic+regeneration+in+mouse+models+of+acute+pancreatitis.">Morphine worsens the severity and prevents pancreatic regeneration in mouse models of acute pancreatitis.</a> Gut. 67 (4), 600-602 (2018).</li><li>Wu, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+systematic+review+of+NSAIDs+treatment+for+acute+pancreatitis+in+animal+studies+and+clinical+trials.">A systematic review of NSAIDs treatment for acute pancreatitis in animal studies and clinical trials.</a> Clinical Research in Hepatology and Gastroenterology. 44, 100002 (2020).</li><li>Schmidt, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+better+model+of+acute+pancreatitis+for+evaluating+therapy.">A better model of acute pancreatitis for evaluating therapy.</a> Annals in Surgery. 215 (1), 44-56 (1992).</li><li>Junyuan, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quercetin+protects+against+intestinal+barrier+disruption+and+inflammation+in+acute+necrotizing+pancreatitis+through+TLR4/MyD88/p38MAPK+and+ERS+inhibition.">Quercetin protects against intestinal barrier disruption and inflammation in acute necrotizing pancreatitis through TLR4/MyD88/p38MAPK and ERS inhibition.</a> Pancreatology. 18 (7), 742-752 (2018).</li><li>Waldron, R. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Orai+Ca(2+)+channel+inhibitor+CM4620+targets+both+parenchymal+and+immune+cells+to+reduce+inflammation+in+experimental+acute+pancreatitis.">The Orai Ca(2+) channel inhibitor CM4620 targets both parenchymal and immune cells to reduce inflammation in experimental acute pancreatitis.</a> Journal of Physiology. 597 (12), 3085-3105 (2019).</li><li>Petersen, O. H., Gerasimenko, J. V., Gerasimenko, O. V., Gryshchenko, O., Peng, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+roles+of+calcium+and+ATP+in+the+physiology+and+pathology+of+the+exocrine+pancreas.">The roles of calcium and ATP in the physiology and pathology of the exocrine pancreas.</a> Physiological Reviews. 101 (4), 1691-1744 (2021).</li></ol>

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</su...

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 the United States, AP is one of the most common reasons for hospitalization, affecting over 200,000 patients5. Moreover, AP, especially SAP, is increasing annually and affecting younger age groups6. However, there is a lack of effective treatment options in current clinical practice6,7. Therefore, it is necessary to explore the molecular mechanisms involved in AP, thereby facilitating treatment improvement.
Well-established experimental animal models are required for studying the mechanisms involved in AP and evaluating the effectiveness of different treatment modalities. With the easy accessibility of transgenic and knockout strains and their small size, which minimizes the doses of drugs required for in vivo evaluation, mice are preferred for AP research. Therefore, several models of AP have been developed in mice8,9.
Working from a mild pancreatitis rat model induced through the intravenous administration of caerulein10, Niederau et al. developed a SAP mouse model presented with acinar cell necrosis induced using the same drug and injection route11. Although this model possesses several advantages, including noninvasiveness, rapid induction, wide reproducibility, and applicability, the major disadvantage is that only a mild form of AP is developed in most cases, thereby limiting its clinical relevance. Alcohol is considered one of the major etiologic factors of AP; however, Foitzik et al. reported that it causes pancreatic injury only when combined with other factors, such as exocrine hyperstimulation12. Moreover, although alcohol-induced AP models developed via different administration routes, and drug doses have been reported13,14,15, their major disadvantage is the difficulty in reproducing them. Intraperitoneal administration of L-arginine can also induce AP in mice16; however, its low clinical relevance hinders its application. Taurocholate, a bile salt, was first proposed by Creutzfeld et al. in 1965 for inducing a condition resembling human AP via pancreatic duct infusion17. Although controversies exist regarding its clinical relevance in pathophysiology18,19, taurocholate-induced pancreatitis remains an indispensable model for SAP.
As this model is simple to realize and is also effective in mice, it is now one of the most used AP models for small animal in vivo studies. Perides et al. employed sodium taurocholate (TC) to induce SAP in mice20, providing insights to understand its pathology. Combined with genetic modification techniques, this model has allowed us to confirm several specific genes involved in AP. For example, Bicozo et al. showed that a knockout of the CD38 gene protected against a model of TC-infusion pancreatitis and attributed the mechanisms to alterations in intracellular Ca2+ signaling21. Fanczal et al. investigated the physiological implication of TRPM2 expression in the plasma membrane of mouse pancreatic acinar and ductal cells, and demonstrated reduced severity of TC-induced SAP in TRPM2 knockout mice22. Furthermore, this model also provides a simple and effective way to test many novel drugs in vivo. For instance, this method enabled validation of the therapeutic effects of caffeine23, dehydrocholic acid24, and various antioxidants and anticoagulants25,26. This evidence demonstrates the versatility of the TC-induced SAP model. Although Wittel et al. described a similar mouse model27, a lack of details on the implementation procedures could result in an inability to reproduce the findings. In this article, we focus on methods using a simple homemade microsyringe and study TC-induced SAP, thereby providing possible guidance not only for further study of the pathogenesis and treatment of AP, but also for a perfectly adaptable experimental method for many other substances.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.5% iodophor</td>
			<td>Shanghai Likang Disinfectant</td>
			<td>310102</td>
			<td>4 mL/mouse</td>
		</tr>
		<tr>
			<td>0.9% sodium chloride</td>
			<td>Sinopharm Group Co., Ltd.</td>
			<td>10019318</td>
			<td>0.8 mL/mouse</td>
		</tr>
		<tr>
			<td>1% Pentobarbital sodium</td>
			<td>Sigma</td>
			<td>P3761</td>
			<td>0.2 -0.25 mL/mouse</td>
		</tr>
		<tr>
			<td>25 &#956;L flat tip Microliter syringe</td>
			<td>Gaoge, Shanghai</td>
			<td>A124019</td>
			<td></td>
		</tr>
		<tr>
			<td>4% Paraformaldehyde</td>
			<td>Beyotime, Nantong, China</td>
			<td>P0099-500ml</td>
			<td></td>
		</tr>
		<tr>
			<td>5% sodium taurocholate (TC)</td>
			<td>Aladdin</td>
			<td>S100834-5g</td>
			<td>10 &#956;L/SAP mouse</td>
		</tr>
		<tr>
			<td>6-0 Sterile nylon microsuture with threaded needle (1/2 circle)</td>
			<td>Cheng-He</td>
			<td>20093</td>
			<td></td>
		</tr>
		<tr>
			<td>75% alcohol</td>
			<td>Sinopharm Group Co., Ltd.</td>
			<td>10009218</td>
			<td>4 mL/mouse</td>
		</tr>
		<tr>
			<td>8-0 Sterile nylon microsuture with threaded needle (3/8 circle)</td>
			<td>Cheng-He</td>
			<td>19064</td>
			<td></td>
		</tr>
		<tr>
			<td>ALT Activity Assay Kit</td>
			<td>EPNK, Anhui, China</td>
			<td>ALT0012</td>
			<td></td>
		</tr>
		<tr>
			<td>Amylase Assay Kit</td>
			<td>EPNK, Anhui, China</td>
			<td>AMY0012</td>
			<td></td>
		</tr>
		<tr>
			<td>Angled small bulldog clamp with 12 mm jaw (3 cm)</td>
			<td>Cheng-He</td>
			<td>HC-X022</td>
			<td></td>
		</tr>
		<tr>
			<td>aspen shavings or shreds for mouse bedding</td>
			<td>Beijing Vital River Laboratory Animal Technology</td>
			<td>VR03015</td>
			<td></td>
		</tr>
		<tr>
			<td>AST Activity Assay Kit</td>
			<td>EPNK, Anhui, China</td>
			<td>AST0012</td>
			<td></td>
		</tr>
		<tr>
			<td>Blood Urea Nitrogen (BUN) Assay Kit</td>
			<td>EPNK, Anhui, China</td>
			<td>BUN0011</td>
			<td></td>
		</tr>
		<tr>
			<td>C57BL/6 mouse</td>
			<td>Beijing Vital River Laboratory Animal Technology</td>
			<td>213</td>
			<td></td>
		</tr>
		<tr>
			<td>Creatine Assay Kit</td>
			<td>EPNK, Anhui, China</td>
			<td>CRE0012</td>
			<td></td>
		</tr>
		<tr>
			<td>Feature microtome blade</td>
			<td>Beyotime, Nantong, China</td>
			<td>E0994</td>
			<td></td>
		</tr>
		<tr>
			<td>Hemostatic Forceps (9.5 cm, Curved)</td>
			<td>JZ, Shanghai Medical Instruments Co. Ltd.</td>
			<td>JC3901</td>
			<td></td>
		</tr>
		<tr>
			<td>Lipase Assay Kit</td>
			<td>Jiancheng, Nanjing, China</td>
			<td>A054-2-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Microtome</td>
			<td>Leica biosystem, Germany</td>
			<td>RM2245</td>
			<td></td>
		</tr>
		<tr>
			<td>Mindray biochemistry analyzer</td>
			<td>Mindray, Shenzhen, China</td>
			<td>BS-420</td>
			<td></td>
		</tr>
		<tr>
			<td>MPO Assay Kit</td>
			<td>Jiancheng, Nanjing, China</td>
			<td>A044-1-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Normal mouse chow</td>
			<td>Trophic, Nantong, China</td>
			<td>LAD 1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate buffered saline</td>
			<td>Beyotime, Nantong, China</td>
			<td>C0221A</td>
			<td></td>
		</tr>
		<tr>
			<td>Straight micro-bulldog clamp with 5 mm jaw (1.5 cm)</td>
			<td>JZ, Shanghai Medical Instruments Co. Ltd.</td>
			<td>W40130</td>
			<td></td>
		</tr>
		<tr>
			<td>Straight or curved forceps (11.0 cm)</td>
			<td>Cheng-He</td>
			<td>HC-X091A or HC-X090A</td>
			<td></td>
		</tr>
		<tr>
			<td>Straight Scissors (10.0 cm)</td>
			<td>Cheng-He, Ningbo, China</td>
			<td>HC-J039102</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermo Scientific Centrifuge</td>
			<td>Thermo Scientific, USA</td>
			<td>Multifuge X1R</td>
			<td></td>
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establishment of a mouse severe acute pancreatitis model using retrograde injection of sodium taurocholate into the biliopancreatic duct

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.

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...

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

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.

The prevalence of acute pancreatitis (AP), especially severe acute pancreatitis (SAP), is increasing in younger age groups annually. However, there is a ...

The overall goal of this video is to establish a mouse model of acute pancreatitis through retrograde injection of sodium taurocholate into biliopancreatic duct. 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 in order not to interfere with the natural course of the disease, which was evidenced by previous literatures and approved by the Animal Ethics Committee of Soochow University. Fast the mouse eight to 12 hours before the surgery. Prepare 5%sodium taurocholate solution filters through a 0.22 microfilter.Autoclave the surgical instruments and materials, including forceps, scissors, blade holder, needle holder, hemostatic clip, vascular clamps, and the sterile drapes. The micro-syringe was sterilized through irradiation. With the mouse, induce a general anesthesia with intraperitoneal injection of 1%pentobarbital sodium solution at a dosage of 50 milligram per kilogram.Fix the mouse in the supine position on an irradiated foam flat plate with the tape. Remove fur of a mouse with a depilation cream from the xiphoid process level down to the connecting line of the bilateral anterior superior iliac spine, with the left and right sides parallel to the front axillary line. Wipe the skin two to three times with 0.5%iodophor and let it dry for one to two minutes each time.Cover the mouse with sterile drapes. Cut open skin layer and then open subcutaneously. Make an approximate two to three centimeters long median abdominal incision.Locate the duodenum from left to right along the stomach. Gently pull the duodenum out and to the left side using surgical forceps and a disinfectant-soaked cotton swab to expose the pancreas, the biliopancreatic duct, and the duodenal papilla. Use 8-0 sutures to pass through the connection between the duodenum and pancreas around the papilla, pull the suture to the left side of the mouse body to fix the duodenum.Gently pull the skin and the lower edge of the liver aside. Clamp the common hepatic duct. Pierce the microinjector into the distal biliopancreatic duct retrogradely.Fix the needle tip near the duodenal papilla with a clamp. Infuse 10 microliters of 5%sodium taurocholate into the biliopancreatic duct at a rate of 5 microliters per minute. Keep the biliopancreatic duct closed for 5 min to achieve a stable pressure.Remove the clamps. Pull out the microinjector, remove the other clamp and fix the suture. Restore the duodenum to the original anatomical position and check the bleeding.Close the wound with 6-0 sutures. Remove the sterile drape. Disinfect the incision with 0.5%iodophor.Recover the animals from anesthesia on a 37 degree Celsius thermal pad. Administer 0.8 milliliter of 0.9%saline subcutaneously for fluid supplementation. At 24 hour post-operation, anesthetize the mouse with pentobarbital sodium using the same dose and injection route.Collect the whole blood from the abdominal aorta and spin at 1, 500 x g for 15 minutes to isolate the serum for serological testing. Collect the pancreas and the lung tissues for histology evaluation. Immerse the pancreas into 4%paraformaldehyde solution for 24 hours.Dehydrate in a series of alcohol concentrations and embed in paraffin. Use a microtome to prepare 5 micro thick pancreas tissue sections. Perform H&E staining for the histological evaluation of the pancreas.The mice were slightly inactive and had weight lost at 24 hours post-operation compared to pre-operation. The survival rates of the control and SAP mice were 100%and 72.7%respectively, at 72 hours post-operation. According to the H&E staining results from pancreas sections, pulpal necrosis of the pancreas tissue could be observed in SAP mice while no significant changes could be found in control mice.Moreover, significantly increased levels of amylase, lipase, pancreas, and lung MPO activity were found in SAP mice compared to control mice. In addition, significantly elevated levels of the liver and kidney indexes including ALT, AST, BUN, and creatinine were also found. Using retrograde injection of sodium taurocholate into biliopancreatic duct, severe acute pancreatitis could be induced in mice.Well exposure of the biliopancreatic duct to ensure the successful puncture. During the puncture, ensure the appropriate angle and force to avoid puncturing of the biliopancreatic duct. TC induced SAP model descried here is a suitable model to study the mechanisms involved in severe acute pancreatitis.It is an economic model using less expensive reagents and this video helps training the novice surgeon faster.

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