1. Rat biliäre Anatomie Die durchschnittliche Gallengang Durchmesser und die Länge war 1 mm und 29 mm. Der durchschnittliche Abstand zwischen den Pylorus und die Papilla betrug 28 mm. Es gibt zwei wesentliche Bauchspeicheldrüsengänge die in den Gallengang offen - die vordere mündet auf der linken Seite und der hinteren mündet auf der rechten Seite. Neben diesen großen Kanälen, eine Reihe von kleineren Kanälen, die alle drei Teile der Bauchspeicheldrüse Drainage direkt öffnen, um den Gallengang. 6 2. Technik Alle Tierexperimente wurden in Übereinstimmung mit den National Institutes of Health Richtlinien für ethische Forschung durchgeführt und wurden von der Institution Research Committee und Animal Care Committee genehmigt. Männliche Sprague-Dawley-Ratten im Alter von 8 Wochen wurden für diese Demonstration verwendet. Die Ratten wurden von Charles River Laboratories in Wilmington, MA, und alle waren mehr als 300 Gramm erhalten. Ratten hatten Zugang zu Standard-Labor-Diät und wurden in Standard-Laborbedingungen untergebracht. Die Ratten wurden zunächst betäubt mit Isofluoran und hielt unter Narkose während der Operation mit Isofluoran durch eine Maske. Ein Mittellinienschnitt von 2 cm wurde ab dem xiphisternum. Nachdem die Faszie eröffnet wurde, die Leberlappen kranial wurden mit einer Sen Retraktor eingefahren, und der Zwölffingerdarm wurde zurückgezogen kaudal mit Q-Tips. Der Gallengang wurde als eine röhrenförmige Struktur in das Duodenum identifiziert. Der Gallengang wurde unter Spannung gesetzt und teilweise aufgeschnitten. Eine sterile 5 cm langen Polyethylen-Rohr mit einem Innendurchmesser von 0,6 mm und einem Außendurchmesser von 0,9 mm verwendet wurde. Ein Ende dieser Röhre war heiß versiegelt und das andere Ende wurde leicht abgeschrägt. Das abgeschrägte Ende wurde dann in den Gallengang weitergegeben. Dies war in Ort mit 8-0 Nylon Fäden sowohl proximal und distal der Eröffnung in den Gallengang gesichert. Die restlichen Rohr zusammen mit dem verschlossenen Ende war in der Bauchhöhle platziert. Die operative Seite wurde mit physiologischer Kochsalzlösung gespült. Der Bauch wurde dann in 2 Schichten geschlossen, mit 4-0 Polyglactin Naht. Die Linea alba wurde mit kontinuierlichen Nähten verschlossen und die unterbrochene dermale Nähte wurden platziert. Hautleim wurde dann auf die Haut aufgetragen. Die Ratten durften unter Standard-Laborbedingungen mit täglich Körpergewicht Überwachung wieder auf ausreichende Flüssigkeitszufuhr zu gewährleisten. Für die Cholestase Umkehr Operation wurden die Ratten mit Isofluoran wie zuvor beschrieben. Der Unterleib war in den vergangenen Inzisionsstelle eröffnet. Das Rohr wurde identifiziert und aus der Bauchhöhle entnommen. Netzadhäsionen um das Rohr frei werden mit stumpfen und scharfen Dissektion. Eine Website in den Zwölffingerdarm wurde dann für eine komfortable Positionierung des Duodenostomie Rohr identifiziert. Eine Tabaksbeutelnaht, 2-3 mm im Durchmesser, wurde dann platziert mit 8-0 Nylon. Das Duodenum wurde dann in diesem Tabaksbeutelnaht eröffnet. Die versiegelte Ende des Rohres wurde dann auf entsprechende Länge geschnitten. Freier Fluss der Galle wurde sichergestellt. Das Ende des Rohres wurde dann in den Zwölffingerdarm gelegt und die Tabaksbeutelnaht wurde verschärft. Der Bauch war bewässert und in Schichten verschlossen. In einer früheren Studie, wobei Choledochoduodenostomie wurde unter Verwendung einer anderen Technik wurden die Ratten 12 Wochen lang überlebte und die Ratten wurden während dieser Zeit gesund. Auch diese Studie bestätigt die Umkehrung der biochemischen und histologischen Veränderungen nach Choledochoduodenostomie in Ratten, die zur Obstruktion des Gallengangs für einen Zeitraum von 2 Wochen. 7

<ol>
	<li>Gouma, D. J., Roughneen, P. T., Kumar, S., Moody, F. G., Rowlands, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+nutritional+status+associated+with+obstructive+jaundice+and+biliary+drainage+in+rats.">Changes in nutritional status associated with obstructive jaundice and biliary drainage in rats.</a> Am J Clin Nutr. 44, 362-369 (1986).</li><li>Ohtsuka, M., Miyazaki, M., Kondo, Y., Nakajima, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neutrophil-mediated+sinusoidal+endothelial+cell+injury+after+extensive+hepatectomy+in+cholestatic+rats.">Neutrophil-mediated sinusoidal endothelial cell injury after extensive hepatectomy in cholestatic rats.</a> Hepatology. 25, 636-641 (1997).</li><li>Abe, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kupffer+cell-derived+interleukin+10+is+responsible+for+impaired+bacterial+clearance+in+bile+duct-ligated+mice.">Kupffer cell-derived interleukin 10 is responsible for impaired bacterial clearance in bile duct-ligated mice.</a> Hepatology. 40, 414-423 (2004).</li><li>Saiki, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preoperative+internal+biliary+drainage+is+superior+to+external+biliary+drainage+in+liver+regeneration+and+function+after+hepatectomy+in+obstructive+jaundiced+rats.">Preoperative internal biliary drainage is superior to external biliary drainage in liver regeneration and function after hepatectomy in obstructive jaundiced rats.</a> Ann Surg. 230, 655-662 (1999).</li><li>Subhas, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protective+effect+of+methylprednisolone+on+warm+ischemia-reperfusion+injury+in+a+cholestatic+rat+liver.">Protective effect of methylprednisolone on warm ischemia-reperfusion injury in a cholestatic rat liver.</a> Am J Surg. 199, 377-380 (2010).</li><li>Kara, M. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+anatomical+study+on+the+rat+pancreas+and+its+ducts+with+emphasis+on+the+surgical+approach.">The anatomical study on the rat pancreas and its ducts with emphasis on the surgical approach.</a> Ann Anat. 187, 105-112 (2005).</li><li>Ryan, C. J., Than, T., Blumgart, L. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Choledochoduodenostomy+in+the+rat+with+obstructive+jaundice.">Choledochoduodenostomy in the rat with obstructive jaundice.</a> J Surg Res. 23, 321-331 (1977).</li></ol>

Cholestase führt zu einer Vielzahl von Veränderungen im Magen-Darm-System. Es wurden Studien durchgeführt suchen die Veränderungen im Immunsystem und Ernährungszustand. 1-3 Cholestase kann die Ergebnisse der Leber-Operationen zu beeinflussen. Es wurden Studien durchgeführt, um die Ergebnisse der Leber-Operationen in der Gegenwart von Cholestase zu verbessern. 4-5 Mit Verbesserungen in Medizin und Forschung, werden weitere Untersuchungen zur Cholestase durchgeführt werden. Daher ist es notwen...

creation of reversible cholestatic rat model

Cholestase ist ein klinischer Zustand häufig von Chirurgen und Gastroenterologen gestoßen. Cholestase kann verschiedene physiologische Veränderungen und Auswirkungen auf den Ernährungszustand und chirurgische Ergebnisse. Studium der pathophysiologischen Veränderungen in der Leber und andere Organe von Bedeutung ist. Verschiedene Studien haben in cholestatischen Ratten-Modellen getan. Wir verwendeten eine reversible cholestatische Rattenmodell in unserer jüngsten Studie, die die Rolle von Methylprednisolon in der Ischämie Reperfusion. Verschiedene Techniken zur Schaffung eines reversible cholestatische Modell beschrieben worden. Erstellung eines reversible cholestatische Rattenmodell kann in Anbetracht der geringeren Größe und einzigartige hepatopancreatobiliary Anatomie in Ratten eine Herausforderung sein. Dieses Video Artikel veranschaulicht die Schaffung eines reversible cholestatische Modell. Dieses Modell kann in verschiedenen Studien, wie z. B. Blick auf die Veränderungen in Ernährung, physiologischen, pathologischen, histologischen und immunologischen Veränderungen im Gastrointestinaltrakt eingesetzt werden. Dieses Modell kann auch verwendet werden, um die Auswirkungen der Cholestase und verschiedene therapeutische Interventionen am wichtigsten hepatischen Operationen zu sehen.

Watch this Scientific Journal Video about Creation of Reversible Cholestatic Rat Model at JoVE.com

Creation of Reversible Cholestatic Rat Model

Cholestase ist ein klinischer Zustand häufig von Chirurgen und Gastroenterologen gestoßen. Erstellung eines reversible cholestatische Rattenmodell kann in Anbetracht der geringeren Größe und einzigartige hepatopancreatobiliary Anatomie in Ratten eine Herausforderung sein. Dieses Video Artikel veranschaulicht die Schaffung eines reversible cholestatische Modell.

Erstellung von Reversible Cholestatische Rattenmodell

Cholestasis is a clinical condition commonly encountered by both surgeons and gastroenterologists. Cholestasis can cause various physiological changes and ...

creation-of-reversible-cholestatic-rat-model

Research

JoVE Journal

Medicine

14.9K Aufrufe.  Providence Hospital and Medical Centers. Cholestase ist ein klinischer Zustand häufig von Chirurgen und Gastroenterologen gestoßen. Erstellung eines reversible cholestatische Rattenmodell kann in Anbetracht der geringeren Größe und einzigartige hepatopancreatobiliary Anatomie in Ratten eine Herausforderung sein. Dieses Video Artikel veranschaulicht die Schaffung eines reversible cholestatische Modell.

Serie: Creation of Reversible Cholestatic Rat Model

Bile Duct Ligation in Mice: Induction of Inflammatory Liver Injury and Fibrosis by Obstructive Cholestasis

In most vertebrates, the liver produces bile that is necessary to emulsify absorbed fats and enable the digestion of lipids in the small intestine as well as to excrete bilirubin and other metabolic products. In the liver, the experimental obstruction of the extrahepatic biliary system initiates a complex cascade of pathological events that leads to cholestasis and inflammation resulting in a strong fibrotic reaction originating from the periportal fields. Therefore, surgical ligation of the common bile duct has become the most commonly used model to induce obstructive cholestatic injury in rodents and to study the molecular and cellular events that underlie these pathophysiological mechanisms induced by inappropriate bile flow. In recent years, different surgical techniques have been described that either allow reconnection or reanastomosis after bile duct ligation (BDL), e.g., partial BDL, or other microsurgical methods for specific research questions. However, the most frequently used model is the complete obstruction of the common bile duct that induces a strong fibrotic response after 21 to 28 days. The mortality rate can be high due to infectious complications or technical inaccuracies. Here we provide a detailed surgical procedure for the BDL model in mice that induce a highly reproducible fibrotic response in accordance to the 3R rule for animal welfare postulated by Russel and Burch in 1959.

Disruption of bile flow results in severe inflammatory cholestatic liver injury with a characteristic time-dependent sequence of morphological alterations. Here we present a protocol for the surgical ligation of the common bile duct in mice that allows to induce a strong fibrotic response after 21 to 28 days.

Gallengang-Ligation in Mice: Induktion von Entzündungsleber-Verletzung und Fibrose von obstruktiver Cholestase

In most vertebrates, the liver produces bile that is necessary to emulsify absorbed fats and enable the digestion of lipids in the small intestine as well ...

Forschung

Medizin

The Dimethylnitrosamine Induced Liver Fibrosis Model in the Rat

Four to six week old, male Wistar rats were used to produce animal models of liver fibrosis. The process requires four weeks of administration of 10 mg/kg dimethylnitrosamine (DMN), given intraperitoneally for three consecutive days per week. Intraperitoneal injections were performed in the fume hood as DMN is a known hepatoxin and carcinogen. The model has several advantages. Firstly, liver changes can be studied sequentially or at particular stages of interest. Secondly, the stage of liver disease can be monitored by measurement of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) enzymes. Thirdly, the severity of liver damage at different stages can be confirmed by sacrifice of animals at designated time points, followed by histological examination of Masson&#39;s Trichome stained liver tissues. After four weeks of DMN dosing, the typical fibrosis score is 5 to 6 on the Ishak scale. The model can be reproduced consistently and has been widely used to assess the efficacy of potential anti-fibrotic agents.

We describe a method to produce an animal model of liver fibrosis in the rat, and assess the degree of fibrosis by histological examination of the liver. The model can be used to study the development of liver disease as well as to test the efficacy of potential anti-fibrotic agents.

Die Dimethylnitrosamin Induced Liver Fibrosis Modell in der Ratte

Four to six week old, male Wistar rats were used to produce animal models of liver fibrosis. The process requires four weeks of administration of 10 mg/kg ...

Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy (ALPPS) Procedure

Recent clinical data support an aggressive surgical approach to both primary and metastatic liver tumors. For some indications, like colorectal liver metastases, the amount of liver tissue left behind after liver resection has become the main limiting factor of resectability of large or multiple liver tumors. A minimal amount of functional tissue is required to avoid the severe complication of post-hepatectomy liver failure, which has high morbidity and mortality. Inducing liver growth of the prospective remnant prior to resection has become more established in liver surgery, either in the form of portal vein embolization by interventional radiologists or in the form of portal vein ligation several weeks prior to resection. Recently, it was shown that liver regeneration is more extensive and rapid, when the parenchymal transection is added to portal vein ligation in a first stage and then, after only one week of waiting, resection performed in a second stage (Associating Liver Partition and Portal vein ligation for Staged hepatectomy = ALPPS). ALPPS has rapidly become popular across the world, but has been criticized for its high perioperative mortality. The mechanism of accelerated and extensive growth induced by this procedure has not been well understood. Animal models have been developed to explore both the physiological and molecular mechanisms of accelerated liver regeneration in ALPPS. This protocol presents a rat model that allows mechanistic exploration of accelerated regeneration.

Rat Model of the Associating Liver Partition and Portal Vein Ligation for Staged Hepatectomy ALPPS Procedure

Inducing rapid liver hypertrophy using Associating Liver Partition and Portal vein ligation for a Staged hepatectomy (ALPPS) has been proposed for resection of borderline resectable liver tumors. This model may elucidate mechanisms involved in rapid hypertrophy and allows testing of drugs that promote or block the acceleration of regeneration.

Rattenmodell der Leber Partition zuordnen und Pfortader Ligation inszenierten Hepatektomie (ALPPS) Verfahren

Recent clinical data support an aggressive surgical approach to both primary and metastatic liver tumors. For some indications, like colorectal liver ...

Invasive Hemodynamic Characterization of the Portal-hypertensive Syndrome in Cirrhotic Rats

This is a detailed protocol describing invasive hemodynamic measurements in cirrhotic rats for the characterization of portal hypertensive syndrome. Portal hypertension (PHT) due to cirrhosis is responsible for the most severe complications in patients with liver disease. The full picture of the portal hypertensive syndrome is characterized by increased portal pressure (PP) due to the increased intrahepatic vascular resistance (IHVR), hyperdynamic circulation, and increased splanchnic blood flow. Progressive splanchnic arterial vasodilation and increased cardiac output with elevated heart rate (HR) but low arterial pressure characterizes the portal hypertensive syndrome.
Novel therapies are currently being developed that aim to decrease PP by either targeting IHVR or increased splanchnic blood flow &#8212; but side effects on systemic hemodynamics may occur. Thus, a detailed characterization of portal venous, splanchnic, and systemic hemodynamic parameters, including measurement of PP, portal venous blood flow (PVBF), mesenteric arterial blood flow, mean arterial pressure (MAP), and HR is needed for preclinical evaluation of the efficacy of novel treatments for PHT. Our video article provides the reader with a structured protocol for performing invasive hemodynamic measurements in cirrhotic rats. In particular, we describe the catheterization of the femoral artery and the portal vein via an ileocolic vein and the measurement of portal venous and splanchnic blood flow via perivascular Doppler-ultrasound flow probes. Representative results of different rat models of PHT are shown.

Here we describe a detailed protocol for invasive measurements of hemodynamic parameters including portal pressure, splanchnic blood flow, and systemic hemodynamics in order to characterize the portal hypertensive syndrome in rats.

Invasive hämodynamische Charakterisierung des Portal-hypertensiven Syndroms in zirrhotischen Ratten

This is a detailed protocol describing invasive hemodynamic measurements in cirrhotic rats for the characterization of portal hypertensive syndrome. ...

Generation of a Rat Model of Acute Liver Failure by Combining 70% Partial Hepatectomy and Acetaminophen

Acute liver failure (ALF) is a clinical condition caused by various etiologies resulting in the loss of metabolic, biochemical, synthesizing, and detoxifying functions of the liver. In most irreversible liver damage cases, orthotropic liver transplant (OLT) remains the only available treatment. To study the therapeutic potential of a treatment for ALF, its prior testing in an animal model of ALF is essential. In the current study, an ALF model in rats was developed by combining 70% partial hepatectomy (PHx) and injections of acetaminophen (APAP) that provides a therapeutic window of 48 h. The median and left lateral lobes of the liver were removed to excise 70% of the liver mass and APAP was given 24 h postsurgically for 2 days. Survival in ALF-induced animals was found to be severely decreased. The development of ALF was confirmed by altered serum levels of the enzymes alanine amino transferase (ALT), aspartate amino transferase (AST), alkaline phosphatase (ALP); changes in prothrombin time (PT); and assessment of the international normalized ratio (INR). Study of the gene expression profile by qPCR revealed an increase in expression levels of genes involved in apoptosis, inflammation, and in the progression of liver injury. Diffused degeneration of hepatocytes and infiltration of immune cells was observed by histological evaluation. The reversibility of ALF was confirmed by the restoration of survival and serum levels of ALT, AST, and ALP after intrasplenic transplantation of syngeneic healthy rat hepatocytes. This model presents a reliable alternative to the available ALF animal models to study the pathophysiology of ALF as well as to evaluate the potential of a novel therapy for ALF. The use of two different approaches also makes it possible to study the combined effect of physical and drug-induced liver injury. The reproducibility and feasibility of current procedure is an added benefit of the model.

The acute liver failure animal model developed in the current study presents a feasible alternative for the study of potential therapies. The current model employs the combined effect of physical and drug-induced hepatic injury and provides a suitable time window to study the potential of novel therapies.

Generierung eines Rattenmodells für akutes Leberversagen durch Kombination von 70% partieller Hepatektomie und Acetaminophen

Acute liver failure (ALF) is a clinical condition caused by various etiologies resulting in the loss of metabolic, biochemical, synthesizing, and ...

A Mouse Model of Chronic Liver Fibrosis for the Study of Biliary Atresia

Biliary atresia (BA) is a fatal disease involving obstructive jaundice, and it is the most common indication for liver transplantation in children. Due to the complex etiology and unknown pathogenesis, there are still no effective drug treatments. At present, the classic BA mouse model induced by rhesus rotavirus (RRV) is the most commonly used model for studying the pathogenesis of BA. This model is characterized by growth retardation, jaundice of the skin and mucosa, clay stools, and dark yellow urine. The histopathology shows severe liver inflammation and obstruction of the intrahepatic and extrahepatic bile ducts, which are similar to the symptoms of human BA. However, the livers of end-stage mice in this model lack fibrosis and cannot fully simulate the characteristics of liver fibrosis in clinical BA. The presented study developed a novel BA mouse model of chronic liver fibrosis by injecting 5-10 &#181;g of anti-Ly6G antibody four times, with gaps of 2 days after each injection. The results showed that some of the mice successfully formed chronic BA with typical fibrosis after the time lapse, meaning these mice represent a suitable animal model for the virus-induced liver fibrosis mechanistic study of BA and a platform for developing future BA treatments.

We established a mouse model of chronic liver fibrosis, which provides a suitable animal model for the virus-induced liver fibrosis mechanistic study of biliary atresia (BA) and a platform for future BA treatments.

Ein Mausmodell der chronischen Leberfibrose zur Untersuchung der Gallengangsatresie

Biliary atresia (BA) is a fatal disease involving obstructive jaundice, and it is the most common indication for liver transplantation in children. Due to ...

Immunologie und Infektion

Sodium Taurocholate Induced Severe Acute Pancreatitis in C57BL/6 Mice

Biliary acute pancreatitis induction by sodium taurocholate infusion has been widely used by the scientific community due to the representation of the human clinical condition and reproduction of inflammatory events corresponding to the onset of clinical biliary pancreatitis. The severity of pancreatic damage can be assessed by measuring the concentration, speed, and volume of the infused bile acid. This study provides an updated checklist of the materials and methods used in the protocol reproduction and shows the main results from this acute pancreatitis (AP) model. Most of the previous publications have limited themselves to reproducing this model in rats. We have applied this method in mice, which provides additional advantages (i.e., the availability of an arsenal of reagents and antibodies for these animals along with the possibility of working with genetically modified strains of mice) that may be relevant to the study. For acute pancreatitis induction in mice, we present a systematic protocol, with a defined dose of 2.5% sodium taurocholate at an infusion speed 10 &#181;L/min for 3 min in C57BL/6 mice that reaches its maximal level of severity within 12 h of induction and highlight results with outcomes that validate the method. With practice and technique, the total estimated time, from the induction of anesthesia to the completion of the infusion, is 25 min per animal.

Animal models for severe acute pancreatitis enable the study of pathophysiological changes at the initial stage, facilitating observation of the evolution of inflammatory events. Here we provide a protocol for the induction of severe acute biliary pancreatitis by retrograde infusion of sodium taurocholate into the pancreatic duct of anesthetized C57BL/6 mice.

Natriumtaurocolat induzierte schwere akute Pankreatitis bei C57BL/6 Mäusen

Biliary acute pancreatitis induction by sodium taurocholate infusion has been widely used by the scientific community due to the representation of the ...

A Mouse Model for Chronic Pancreatitis via Bile Duct TNBS Infusion

Chronic pancreatitis (CP) is a complex disease involving pancreatic inflammation and fibrosis, glandular atrophy, abdominal pain and other symptoms. Several rodent models have been developed to study CP, of which the bile duct 2,4,6 -trinitrobenzene sulfonic acid (TNBS) infusion model replicates the features of neuropathic pain seen in CP. However, bile duct drug infusion in mice is technically challenging. This protocol demonstrates the procedure of bile duct TNBS infusion for generation of a CP mouse model. TNBS was infused into the pancreas through the ampulla of Vater in the duodenum. This protocol optimized drug volume, surgical techniques, and drug handling during the procedure. TNBS-treated mice showed features of CP as reflected by bodyweight and pancreas weight reductions, changes in pain-associated behaviors, and abnormal pancreatic morphology. With these improvements, mortality associated with TNBS injection was minimal. This procedure is not only critical in generating pancreatic disease models but is also useful in local pancreatic drug delivery.

Chronic pancreatitis (CP) is a disease characterized by inflammation and fibrosis of the pancreas, often associated with intractable abdominal pain. This article focuses on refining the technique to generate a mouse model of CP via bile duct infusion with 2,4,6 -trinitrobenzene sulfonic acid (TNBS).

Ein Mausmodell für chronische Pankreatitis mittels Gallengangs-TNBS-Infusion

Chronic pancreatitis (CP) is a complex disease involving pancreatic inflammation and fibrosis, glandular atrophy, abdominal pain and other symptoms. ...

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.

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.

Etablierung eines Mausmodells für schwere akute Pankreatitis mittels retrograder Injektion von Natriumtaurocholat in den Gallengang

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

The Murine Choline-Deficient, Ethionine-Supplemented (CDE) Diet Model of Chronic Liver Injury

Chronic liver diseases, such as viral hepatitis, alcoholic liver disease, or non-alcoholic fatty liver disease, are characterized by continual inflammation, progressive destruction and regeneration of the hepatic parenchyma, liver progenitor cell proliferation, and fibrosis. The end-stage of every chronic liver disease is cirrhosis, a major risk factor for the development of hepatocellular carcinoma. To study processes regulating disease initiation, establishment, and progression, several animal models are used in laboratories. Here we describe a six-week time course of the choline-deficient and ethionine-supplemented (CDE) mouse model, which involves feeding six-week old male C57BL/6J mice with choline-deficient chow and 0.15% DL-ethionine-supplemented drinking water. Monitoring of animal health and a typical body weight loss curve are explained. The protocol demonstrates the gross examination of a CDE-treated liver and blood collection by cardiac puncture for subsequent serum analyses. Next, the liver perfusion technique and collection of different hepatic lobes for standard evaluations are shown, including liver histology assessments by hematoxylin and eosin or Sirius Red stainings, immunofluorescent detection of hepatic cell populations as well as transcriptome profiling of the liver microenvironment. This mouse model is suitable for studying inflammatory, fibrogenic, and liver progenitor cell dynamics induced through chronic liver disease and can be used to test potential therapeutic agents that may modulate these processes.

The Murine Choline-Deficient, Ethionine-Supplemented CDE Diet Model of Chronic Liver Injury

Here we describe a common method to induce chronic liver injury in mice by feeding of a choline-deficient and ethionine-supplemented (CDE) diet. We demonstrate health monitoring, liver perfusion, isolation, and preservation. A time course of six weeks can inform about liver injury, pathohistology, fibrosis, inflammatory, and liver progenitor cell responses.