Eipel C, Kidess E, Abshagen K, Leminh K, Menger M D, Burkhardt H, Vollmar B
Institute for Experimental Surgery, University of Rostock, Rostock, Germany.
Br J Pharmacol. 2007 Jun;151(3):406-13. doi: 10.1038/sj.bjp.0707230. Epub 2007 Apr 10.
There is major evidence for the strong bi-directional interrelation of parenchymal cell apoptosis and leukocyte accumulation and inflammation in acute liver injury. Therefore, the aim of this in vivo study was to investigate the anti-apoptotic and anti-inflammatory potential of antileukoproteinase (ALP) in a murine model of acute liver failure.
C57BL/6J mice were given galactosamine (D-GalN) and E. coli lipopolysaccharide (LPS) followed by administration of saline or ALP. Besides survival rate, hepatic tissue damage and inflammatory response were analyzed by intravital fluorescence microscopy 6 hours after treatment. In addition, immunohistochemical analysis of NFkappaB-p65 and hepatocellular apoptosis, plasma levels of AST/ALT, TNF-alpha and IL-10 were determined.
Administration of D-GalN/LPS provoked hepatic damage, including marked leukocyte recruitment and microvascular perfusion failure, as well as hepatocellular apoptosis and enzyme release. NFkappaB-p65 became increasingly detectable in hepatocellular nuclei, accompanied by a rise of TNF-alpha and IL-10 plasma levels. ALP markedly reduced intrahepatic leukocyte accumulation, nuclear translocation of NFkappaB and plasma levels of TNF-alpha and IL-10. Moreover, liver enzyme levels indicated the absence of necrotic parenchymal cell death. In contrast, ALP failed to block both apoptosis and caspase-3 levels and the mortality rate of ALP-treated animals was comparable to that of saline-treated mice.
ALP could effectively prevent D-GalN/LPS-associated intrahepatic inflammatory responses by inhibition of NFkappaB activity, but not apoptosis-driven mortality. Thus, a protease-inactivating approach such as application of ALP seems to be inadequate in damaged liver where apoptosis represents the predominant mode of cell death.
有大量证据表明，在急性肝损伤中，实质细胞凋亡与白细胞聚集及炎症之间存在强烈的双向相互关系。因此，本体内研究旨在探讨抗白细胞蛋白酶（ALP）在急性肝衰竭小鼠模型中的抗凋亡和抗炎潜力。
给C57BL/6J小鼠注射半乳糖胺（D - GalN）和大肠杆菌脂多糖（LPS），随后给予生理盐水或ALP。除存活率外，在治疗6小时后通过活体荧光显微镜分析肝组织损伤和炎症反应。此外，测定NFκB - p65的免疫组化分析、肝细胞凋亡、血浆中AST/ALT、TNF - α和IL - 10水平。
给予D - GalN/LPS可引发肝损伤，包括显著的白细胞募集和微血管灌注衰竭，以及肝细胞凋亡和酶释放。NFκB - p65在肝细胞核中越来越容易检测到，同时血浆中TNF - α和IL - 10水平升高。ALP显著减少肝内白细胞聚集、NFκB的核转位以及血浆中TNF - α和IL - 10水平。此外，肝酶水平表明不存在坏死性实质细胞死亡。相比之下，ALP未能阻止凋亡和caspase - 3水平，且接受ALP治疗的动物死亡率与接受生理盐水治疗的小鼠相当。
ALP可通过抑制NFκB活性有效预防D - GalN/LPS相关的肝内炎症反应，但不能预防凋亡驱动的死亡率。因此，在凋亡是主要细胞死亡模式的受损肝脏中，应用ALP等蛋白酶失活方法似乎并不充分。
