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Use of a Hanging-weight System for Liver Ischemia in Mice

机译:吊重系统在小鼠肝脏缺血中的应用

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摘要

Acute liver injury due to ischemia can occur during several clinical procedures e.g. liver transplantation, hepatic tumor resection or trauma repair and can result in liver failure which has a high mortality rate1-2. Therefore murine studies of hepatic ischemia have become an important field of research by providing the opportunity to utilize pharmacological and genetic studies3-9. Specifically, conditional mice with tissue specific deletion of a gene (cre, flox system) provide insights into the role of proteins in particular tissues10-13 . Because of the technical difficulty associated with manually clamping the portal triad in mice, we performed a systematic evaluation using a hanging-weight system for portal triad occlusion which has been previously described3. By using a hanging-weight system we place a suture around the left branch of the portal triad without causing any damage to the hepatic lobes, since also the finest clamps available can cause hepatic tissue damage because of the close location of liver tissue to the vessels. Furthermore, the right branch of the hepatic triad is still perfused thus no intestinal congestion occurs with this technique as blood flow to the right hepatic lobes is preserved. Furthermore, the portal triad is only manipulated once throughout the entire surgical procedure. As a result, procedures like pre-conditioning, with short times of ischemia and reperfusion, can be easily performed. Systematic evaluation of this model by performing different ischemia and reperfusion times revealed a close correlation of hepatic ischemia time with liver damage as measured by alanine (ALT) and aspartate (AST) aminotransferase serum levels3,9. Taken together, these studies confirm highly reproducible liver injury when using the hanging-weight system for hepatic ischemia and intermittent reperfusion. Thus, this technique might be useful for other investigators interested in liver ischemia studies in mice. Therefore the video clip provides a detailed step-by-step description of this technique.
机译:由缺血引起的急性肝损伤可能在几种临床程序中发生,例如肝移植,肝肿瘤切除或创伤修复可能导致肝功能衰竭,死亡率高 1-2 。因此,通过提供利用药理和遗传学研究的机会 3-9 ,肝缺血的鼠类研究已成为重要的研究领域。具体来说,具有组织特异性缺失基因(cre,flox系统)的条件小鼠可以深入了解蛋白质在特定组织中的作用 10-13 。由于与手动夹紧小鼠的三叉戟相关的技术难度,我们使用悬挂重量系统对三叉戟进行闭塞进行了系统的评估,这已经在前面进行了描述 3 。通过使用悬挂式砝码系统,我们在门三联征的左分支周围放置了一条缝合线,而不会对肝叶造成任何损害,因为由于肝脏组织与血管的位置靠近,因此可用的最精细的夹具也会造成肝组织损害。此外,肝三联征的右分支仍被灌注,因此该技术不会引起肠道充血,因为保留了流向右肝叶的血流。此外,在整个外科手术过程中仅对门部三重轴进行一次操作。结果,可以容易地进行诸如缺血短时间和再灌注的预处理之类的程序。通过进行不同的缺血和再灌注时间对该模型进行系统评价,结果显示,通过丙氨酸(ALT)和天冬氨酸(AST)氨基转移酶血清水平 3,9 测得的肝缺血时间与肝脏损害密切相关。综上所述,这些研究证实了使用悬挂式重物系统进行肝缺血和间歇性再灌注时,肝损伤具有高度可再现性。因此,该技术对于对小鼠肝缺血研究感兴趣的其他研究者可能有用。因此,视频剪辑提供了此技术的详细分步说明。

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