PDF(Pubmed)
Abstract：
Alcohol-related liver disease (ALD) encompasses a range of pathological conditions that are complex to study at the clinical and preclinical levels. Despite the global burden of ALD, there is a lack of effective treatments, and mortality is high. One of the reasons for the unsuccessful development of novel therapies is that experimental studies are hindered by the challenge of recapitulating this multifactorial disorder in vitro, including the contributions of hepatotoxicity, impaired lipid metabolism, fibrosis and inflammatory cytokine storm, which are critical drivers in the pathogenesis of ALD in patients and primary targets for drug development. Here, we present the unique characteristics of the culture of human precision-cut liver slices (PCLS) to replicate key disease processes in ALD. PCLS were prepared from human liver specimens and treated with ethanol alone or in combination with fatty acids and lipopolysaccharide (FA + LPS) for up to 5 days to induce hepatotoxic, inflammatory and fibrotic events associated with ALD. Alcohol insult induced hepatocyte death which was more pronounced with the addition of FA + LPS. This mixture showed a significant increase in the cytokines conventionally associated with the prototypical inflammatory response observed in severe ALD, and interestingly, alcohol alone exhibited a different effect. Profibrogenic activation was also observed in the slices and investigated in the context of slice preparation. These results support the versatility of this organotypic model to study different pathways involved in alcohol-induced liver damage and ALD progression and highlight the applicability of the PCLS for drug discovery, confirming their relevance as a bridge between preclinical and clinical studies.
摘要：
酒精相关性肝病（ALD）包括一系列复杂的临床和临床前水平研究的病理状况。尽管ALD的全球负担，缺乏有效的治疗方法，死亡率很高。新疗法开发失败的原因之一是，在体外概括这种多因素疾病的挑战阻碍了实验研究，包括肝毒性的贡献，脂质代谢受损，纤维化和炎性细胞因子风暴，这些是患者ALD发病机制的关键驱动因素和药物开发的主要目标。这里,我们提出了人类精确切割肝片（PCLS）培养的独特特征，以复制ALD中的关键疾病过程。PCLS从人肝标本制备，用乙醇单独或与脂肪酸和脂多糖（FA+LPS）组合处理长达5天，以诱导肝毒性，与ALD相关的炎症和纤维化事件。酒精损伤诱导肝细胞死亡，这在添加FALPS时更为明显。该混合物显示与在严重ALD中观察到的典型炎症反应常规相关的细胞因子显著增加。有趣的是，单独的酒精表现出不同的效果。在切片中也观察到促纤维化活化,并在切片制备的背景下进行研究。这些结果支持了这种器官模型的多功能性，以研究涉及酒精诱导的肝损伤和ALD进展的不同途径，并强调了PCLS在药物发现中的适用性。确认它们作为临床前和临床研究之间的桥梁的相关性。
