Development of an Ethanol-induced Fibrotic Liver Model in Zebrafish to Study Progenitor Cell-mediated Hepatocyte Regeneration.
Development of an Ethanol-induced Fibrotic Liver Model in Zebrafish to Study Progenitor Cell-mediated Hepatocyte Regeneration.
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DOI：
10.3791/54002
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发表时间：
2016-05
期刊：
影响因子：
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通讯作者：
Mianbo Huang;Jin Xu;C. Shin
中科院分区：
文献类型：
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作者：
Mianbo Huang;Jin Xu;C. Shin
持续的肝纤维化伴随细胞外基质(ECM)蛋白的持续积聚导致肝脏细胞功能的丧失，导致肝硬变并肝细胞功能障碍。在多种肝脏侮辱中，酗酒会导致严重的健康问题，包括肝功能衰竭和肝细胞癌。尽管如此，再生肝细胞的内源性细胞来源对酒精的反应还没有得到适当的研究。此外，很少有研究有效地将肝细胞再生模拟在酒精诱导的损伤上。我们最近报道建立了乙醇(EtoH)诱导的斑马鱼肝纤维化模型，在该模型中，肝祖细胞(HPC)在肝细胞几乎完全丧失的情况下产生肝细胞。此外，通过使用该模型的化学筛选，我们确定了多个促进肝细胞再生的小分子。在这里，我们详细描述了建立乙醇诱导的肝纤维化模型和在斑马鱼中使用该模型进行化学筛选的过程。这一方案将是描述肝细胞如何在纤维化肝脏中再生的分子和细胞机制的关键工具。此外，这些方法将有助于在体内发现新的慢性肝病治疗策略。
Sustained liver fibrosis with continuation of extracellular matrix (ECM) protein build-up results in the loss of cellular competency of the liver, leading to cirrhosis with hepatocellular dysfunction. Among multiple hepatic insults, alcohol abuse can lead to significant health problems including liver failure and hepatocellular carcinoma. Nonetheless, the identity of endogenous cellular sources that regenerate hepatocytes in response to alcohol has not been properly investigated. Moreover, few studies have effectively modeled hepatocyte regeneration upon alcohol-induced injury. We recently reported on establishing an ethanol (EtOH)-induced fibrotic liver model in zebrafish in which hepatic progenitor cells (HPCs) gave rise to hepatocytes upon near-complete hepatocyte loss in the presence of fibrogenic stimulus. Furthermore, through chemical screens using this model, we identified multiple small molecules that enhance hepatocyte regeneration. Here we describe in detail the procedures to develop an EtOH-induced fibrotic liver model and to perform chemical screens using this model in zebrafish. This protocol will be a critical tool to delineate the molecular and cellular mechanisms of how hepatocyte regenerates in the fibrotic liver. Furthermore, these methods will facilitate potential discovery of novel therapeutic strategies for chronic liver disease in vivo.