PDF(Pubmed)
Abstract：
Acute alcoholic hepatitis (AAH) from binge drinking is a serious disease. It is associated with a high mortality rate, especially among young adults. Apoptosis is known to be a primary cause of liver damage, and it can be induced by either intrinsic signaling pathways or by reactive oxygen species (ROS). Adenosine A1 receptors (ADORA1) are known to be involved in ethanol metabolism; however, underlying mechanism is not well understood. For investigating how the intrinsic ADORA1 function in ethanol metabolism in normal human hepatocytes without interference by extrinsic molecules, primary hepatocytes pose a challenge, due to unavoidable contamination by other kinds of cells in the liver. Also, they are difficult to culture stably. As a novel alternative, hepatocytes derived from human-induced pluripotent stem cells were employed because they display similar function to primary hepatocytes and they can be stably cultured. The dynamics and integrity of signal transduction mechanisms were investigated by following chronological changes in gene expression. This shed light on how and when the ADORA1 function and on causal relationships between the pathways and clinical symptoms. The findings of the present study shows that ADORA1 are most activated soon after exposure to ethanol, and transfection of small interfering RNA targeting ADORA1-messenger-RNA (ADORA1-siRNA) into the hepatocytes significantly suppresses production of actin protein and ROS. It suggests that ADORA1 in the liver contribute to apoptosis in acute alcoholism through both intrinsic pathway and ROS activity. Also, actin that is abundant in the cells could be an appropriate biomarker evaluating hepatic function status.
摘要：
暴饮暴食引起的急性酒精性肝炎（AAH）是一种严重的疾病。它与高死亡率有关，尤其是年轻人。细胞凋亡是肝损伤的主要原因，它可以由内在信号通路或活性氧（ROS）诱导。已知腺苷A1受体(ADORA1)参与乙醇代谢;然而,潜在的机制还没有很好的理解。为了研究内在ADORA1在正常人肝细胞的乙醇代谢中如何发挥作用，而不受外在分子的干扰，原代肝细胞构成挑战，由于肝脏中其他种类的细胞不可避免地污染。此外，它们很难稳定培养。作为一种新颖的选择，采用源自人诱导多能干细胞的肝细胞,因为它们显示与原代肝细胞相似的功能并且它们可以稳定培养。通过跟踪基因表达的时间变化，研究了信号转导机制的动力学和完整性。这揭示了ADORA1如何以及何时起作用以及途径与临床症状之间的因果关系。本研究的结果表明，ADORA1在暴露于乙醇后不久被激活，靶向ADORA1-信使RNA（ADORA1-siRNA）的小干扰RNA转染到肝细胞中显着抑制肌动蛋白和ROS的产生。这表明肝脏中的ADORA1通过内在途径和ROS活性促进急性酒精中毒中的细胞凋亡。此外，在细胞中丰富的肌动蛋白可能是评估肝功能状态的适当生物标志物。
