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A dynamic plasma membrane proteome analysis of alcohol-induced liver cirrhosis

机译:酒精性肝硬化的动态质膜蛋白质组学分析

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摘要

Alcohol-induced injury has become one of the major causes for liver cirrhosis. However, the molecular mechanisms of ethanol-induced injury are not fully understood. To this end, we performed a dynamic plasma membrane proteomic research on rat model. A rat model from hepatitis to liver cirrhosis was developed. Plasma membrane from liver tissue with liver fibrosis stage of 2 and 4 (S2 and S4) was purified by sucrose density gradient centrifugation. Its purification was verified by western blotting. Proteins from plasma membrane were separated by two-dimensional electrophoresis (2DE) and differentially expressed proteins were identified by tandem mass spectrometry. 16 consistent differentially expressed proteins from S2 to S4 were identified by mass spectrometry. The expression of differentially expressed proteins annexin A6 and annexin A3 were verified by western blotting, and annexin A3 was futher verified by immunohistochemistry. Our research suggests a possible mechanism by which ethanol alters protein expression to enhance the liver fibrosis progression. These differentially expressed proteins might be new drug targets for treating alcoholic liver cirrhosis.
机译:酒精引起的伤害已成为肝硬化的主要原因之一。但是,乙醇诱导的损伤的分子机制尚不完全清楚。为此,我们对大鼠模型进行了动态质膜蛋白质组学研究。建立了从肝炎到肝硬化的大鼠模型。通过蔗糖密度梯度离心纯化肝纤维化阶段为2和4(S2和S4)的肝组织的质膜。通过蛋白质印迹证实了其纯化。通过二维电泳(2DE)分离质膜中的蛋白质,并通过串联质谱法鉴定差异表达的蛋白质。通过质谱鉴定了从S2到S4的16个一致的差异表达蛋白。通过蛋白质印迹法验证差异表达蛋白膜联蛋白A6和膜联蛋白A3的表达,并通过免疫组织化学进一步验证膜联蛋白A3。我们的研究提出了乙醇可能通过改变蛋白质表达来增强肝纤维化进程的可能机制。这些差异表达的蛋白质可能是治疗酒精性肝硬化的新药物靶标。

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