Ellis Jillian L, Yin Chunyue
Division of Gastroenterology, Hepatology and Nutrition, Cincinnati Children's Hospital Medical Center.
Division of Gastroenterology, Hepatology and Nutrition, Cincinnati Children's Hospital Medical Center; Division of Developmental Biology, Cincinnati Children's Hospital Medical Center;
J Vis Exp. 2017 May 25(123):55630. doi: 10.3791/55630.
Alcoholic Liver Disease (ALD) refers to damage to the liver due to acute or chronic alcohol abuse. It is among the leading causes of alcohol-related morbidity and mortality and affects more than 2 million people in the United States. A better understanding of the cellular and molecular mechanisms underlying alcohol-induced liver injury is crucial for developing effective treatment for ALD. Zebrafish larvae exhibit hepatic steatosis and fibrogenesis after just 24 h of exposure to 2% ethanol, making them useful for the study of acute alcoholic liver injury. This work describes the procedure for acute ethanol treatment in zebrafish larvae and shows that it causes steatosis and swelling of the hepatic blood vessels. A detailed protocol for Hematoxylin and Eosin (H&E) staining that is optimized for the histological analysis of the zebrafish larval liver, is also described. H&E staining has several unique advantages over immunofluorescence, as it marks all liver cells and extracellular components simultaneously and can readily detect hepatic injury, such as steatosis and fibrosis. Given the increasing usage of zebrafish in modeling toxin and virus-induced liver injury, as well as inherited liver diseases, this protocol serves as a reference for the histological analyses performed in all these studies.
酒精性肝病（ALD）是指由于急性或慢性酒精滥用导致的肝脏损伤。它是与酒精相关的发病和死亡的主要原因之一，在美国影响着超过200万人。更好地了解酒精性肝损伤的细胞和分子机制对于开发有效的ALD治疗方法至关重要。斑马鱼幼体在暴露于2%乙醇仅24小时后就会出现肝脂肪变性和纤维化，这使得它们可用于急性酒精性肝损伤的研究。这项工作描述了斑马鱼幼体急性乙醇处理的程序，并表明它会导致肝血管脂肪变性和肿胀。还描述了一种针对斑马鱼幼体肝脏组织学分析优化过的苏木精和伊红（H&E）染色详细方案。H&E染色相对于免疫荧光有几个独特的优点，因为它能同时标记所有肝细胞和细胞外成分，并且能很容易地检测到肝损伤，如脂肪变性和纤维化。鉴于斑马鱼在模拟毒素和病毒诱导的肝损伤以及遗传性肝病方面的使用越来越多，该方案可作为所有这些研究中进行组织学分析的参考。
