Laboratoire d'Hépatologie Cellulaire, Centre de Recherche du Centre Hospitalier de l'Université de Montréal, Université de Montréal, Montreal, Quebec, Canada.
Département de Médecine, Université de Montréal, Montreal, Quebec, Canada.
Liver Transpl. 2019 Apr;25(4):627-639. doi: 10.1002/lt.25415. Epub 2019 Mar 1.
Liver ischemia/reperfusion injury (IRI) is an important cause of liver damage especially early after liver transplantation, following liver resection, and in other clinical situations. Using rat experimental models, we identified oxaloacetate (OAA) as a key metabolite able to protect hepatocytes from hypoxia and IRI. In vitro screening of metabolic intermediates beneficial for hepatocyte survival under hypoxia was performed by measures of cell death and injury. In vivo, the effect of OAA was evaluated using the left portal vein ligation (LPVL) model of liver ischemia and a model of warm IRI. Liver injury was evaluated in vivo by serum transaminase levels, liver histology, and liver weight (edema). Levels and activity of caspase 3 were also measured. In vitro, the addition of OAA to hepatocytes kept in a hypoxic environment significantly improved cell viability (P < 0.01), decreased cell injury (P < 0.01), and improved energy metabolism (P < 0.01). Administration of OAA significantly reduced the extent of liver injury in the LPVL model with lower levels of alanine aminotransferase (ALT; P < 0.01), aspartate aminotransferase (AST; P < 0.01), and reduced liver necrosis (P < 0.05). When tested in a warm IRI model, OAA significantly decreased ALT (P < 0.001) and AST levels (P < 0.001), prevented liver edema (P < 0.001), significantly decreased caspase 3 expression (P < 0.01), as well as histological signs of cellular vesiculation and vacuolation (P < 0.05). This was associated with higher adenosine triphosphate (P < 0.05) and energy charge levels (P < 0.01). In conclusion, OAA can significantly improve survival of ischemic hepatocytes. The hepatoprotective effect of OAA was associated with increased levels of liver bioenergetics both in vitro and in vivo. These results suggest that it is possible to support mitochondrial activity despite the presence of ischemia and that OAA can effectively reduce ischemia-induced injury in the liver.
肝缺血/再灌注损伤(IRI)是肝损伤的一个重要原因,特别是在肝移植后、肝切除后和其他临床情况下。我们使用大鼠实验模型发现草酰乙酸(OAA)是一种关键代谢物,能够保护肝细胞免受缺氧和 IRI 的损伤。通过细胞死亡和损伤的测量,在体外筛选对缺氧下肝细胞存活有益的代谢中间产物。在体内,通过左门静脉结扎(LPVL)肝缺血模型和热 IRI 模型来评估 OAA 的作用。通过血清转氨酶水平、肝组织学和肝重量(水肿)评估体内肝损伤。还测量了半胱天冬酶 3 的水平和活性。在体外,将 OAA 添加到处于缺氧环境中的肝细胞中,可显著提高细胞活力(P < 0.01),降低细胞损伤(P < 0.01),并改善能量代谢(P < 0.01)。OAA 的给药显著降低了 LPVL 模型中肝损伤的程度,使丙氨酸氨基转移酶(ALT;P < 0.01)、天冬氨酸氨基转移酶(AST;P < 0.01)水平降低,肝坏死减少(P < 0.05)。当在热 IRI 模型中进行测试时,OAA 显著降低 ALT(P < 0.001)和 AST 水平(P < 0.001),防止肝水肿(P < 0.001),显著降低半胱天冬酶 3 表达(P < 0.01),以及细胞空泡化和空泡化的组织学迹象(P < 0.05)。这与三磷酸腺苷(ATP;P < 0.05)和能量电荷水平(P < 0.01)的升高有关。总之,OAA 可以显著提高缺血肝细胞的存活率。OAA 的肝保护作用与体外和体内的肝生物能水平的提高有关。这些结果表明,尽管存在缺血,支持线粒体活性是可能的,并且 OAA 可以有效地减少肝脏的缺血性损伤。