Plasma membrane proteome analysis of the early effect of alcohol on liver: implications for alcoholic liver disease
Plasma membrane proteome analysis of the early effect of alcohol on liver: implications for alcoholic liver disease
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DOI：
10.1093/abbs/gmq108
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发表时间：
2011-01-01
影响因子：
3.7
通讯作者：
Yuan, Zhenghong
中科院分区：
文献类型：
--
作者：
Zhang, Lijun;Jia, Xiaofang;Yuan, Zhenghong
在人类身上，过度饮酒会导致严重的肝病。为了检验酒精对肝病的早期影响，给大鼠足够的酒精以发展为肝硬变。在这项工作中，我们研究了纤维化发生前的大鼠。两次蔗糖密度梯度离心法提取肝细胞质膜。用双向凝胶电泳法(2-DE)和相对和绝对定量等压标签(ITRAQ)技术分析乙醇处理大鼠和对照组PM的蛋白质组图谱。乙醇处理改变了15种不同的肝脏蛋白质的含量：其中10种用2-DE检测到，5种用iTRAQ检测到。两种方法均检测角蛋白8。对这些差异检测到的蛋白质进行的基因本体论分析表明，它们大多参与了重要的细胞功能，如结合活性(包括离子、DNA、ATP结合等)、细胞结构或酶活性。其中，膜联蛋白A2、角蛋白8和角蛋白18用免疫印迹分析进一步证实，膜联蛋白A2用免疫组织化学方法证实。我们的结果表明，酒精可能通过改变PM中几个相关蛋白质的表达水平来影响细胞结构、黏附和酶活性据我们所知，这是首次研究酒精对肝脏PM蛋白质组的影响，这可能有助于了解酒精性肝病的可能机制。
In humans, the over-consumption of alcohol can lead to serious liver disease. To examine the early effects of alcohol on liver disease, rats were given sufficient ethanol to develop liver cirrhosis. Rats before the onset of fibrosis were studied in this work. Plasma membranes (PM) of liver were extracted by twice sucrose density gradient centrifugation. The proteome profiles of PM from ethanol-treated rats and the controls were analyzed using two-dimensional gel electrophoresis (2-DE) and isobaric tag for relative and absolute quantitation (iTRAQ) technology. Ethanol treatment altered the amount of 15 different liver proteins: 10 of them were detected by 2-DE and 5 by iTRAQ. Keratin 8 was detected by both methods. Gene ontology analysis of these differentially detected proteins indicated that most of them were involved in important cell functions such as binding activity (including ion, DNA, ATP binding, etc.), cell structure, or enzyme activity. Among these, annexin A2, keratin 8, and keratin 18 were further verified using western blot analysis and annexin A2 was verified by immunohistochemistry. Our results suggested that alcohol has the potential to affect cell structure, adhesion and enzyme activity by altering expression levels of several relevant proteins in the PM. To the best of our knowledge, this is the first time to study the effect of alcohol on the liver PM proteome and it might be helpful for understanding the possible mechanisms of alcohol-induced liver disease.