Tag Carmen G, Sauer-Lehnen Sibille, Weiskirchen Sabine, Borkham-Kamphorst Erawan, Tolba René H, Tacke Frank, Weiskirchen Ralf
Institute of Molecular Pathobiochemistry, Experimental Gene Therapy and Clinical Chemistry, RWTH Aachen University.
Institute for Laboratory Animal Science and Experimental Surgery, RWTH Aachen University.
J Vis Exp. 2015 Feb 10(96):52438. doi: 10.3791/52438.
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.
在大多数脊椎动物中,肝脏产生胆汁,胆汁对于乳化吸收的脂肪、促进小肠中脂质的消化以及排泄胆红素和其他代谢产物是必需的。在肝脏中,肝外胆道系统的实验性梗阻会引发一系列复杂的病理事件,导致胆汁淤积和炎症,进而引发源自门周区域的强烈纤维化反应。因此,胆总管的外科结扎已成为诱导啮齿动物梗阻性胆汁淤积性损伤以及研究由异常胆汁流动引起的这些病理生理机制背后的分子和细胞事件的最常用模型。近年来,已经描述了不同的外科技术,例如部分胆管结扎(BDL),或者用于特定研究问题的其他显微外科方法,这些技术在胆管结扎后允许重新连接或吻合。然而,最常用的模型是胆总管完全梗阻,在21至28天后会引发强烈的纤维化反应。由于感染性并发症或技术失误,死亡率可能会很高。在此,我们提供了一种用于小鼠BDL模型的详细手术方法,该方法根据1959年拉塞尔和伯奇提出的动物福利3R原则,可诱导高度可重复的纤维化反应。