Analysis of the liver mitochondrial proteome in response to ethanol and S-adenosylmethionine treatments: novel molecular targets of disease and hepatoprotection
Analysis of the liver mitochondrial proteome in response to ethanol and S-adenosylmethionine treatments: novel molecular targets of disease and hepatoprotection
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DOI：
10.1152/ajpgi.00332.2009
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发表时间：
2010-05-01
影响因子：
4.5
通讯作者：
Bailey, Shannon M.
中科院分区：
文献类型：
--
作者：
Andringa, Kelly K.;King, Adrienne L.;Bailey, Shannon M.
安德林加KK，国王AL，埃克莱斯顿HB，曼特纳SK，兰达尔A，贾拉NC，迪金森DA，斯夸德里托GL，贝利SM。乙醇和S-腺苷甲硫氨酸治疗后肝线粒体蛋白质组分析：疾病和肝保护的新分子靶点。美国生理学杂志胃肠和肝脏生理学298：G732-G745，2010。首次发表于2010年2月11日; doi：10.1152/ajpgi.00332.2009。S-腺苷甲硫氨酸（SAM）最大限度地减少酒精肝毒性，然而，负责SAM肝保护的分子机制仍然未知。在此，我们使用蛋白质组学来确定SAM对早期酒精性肝病的肝脏保护作用是否与线粒体蛋白质组的改变有关。为此，给雄性大鼠喂食对照或含乙醇的流质饮食+/- SAM，并制备肝线粒体用于蛋白质组学分析。二维等电聚焦（2D IEF/SDS-PAGE）和蓝色非变性凝胶电泳（BN-PAGE）分别用于确定基质和氧化磷酸化（OxPhos）蛋白的变化。SAM共施用使酒精依赖性炎症最小化并保护线粒体呼吸。SAM补充乙醇喂养大鼠的肝脏SAM水平，然而，线粒体SAM水平增加乙醇和SAM治疗。使用2D IEF/SDS-PAGE，30个蛋白质在响应于乙醇、SAM或两者的丰度上显示出显著变化。受乙醇和SAM处理影响的蛋白质类是分子伴侣、β氧化蛋白、硫代谢蛋白和参与甲硫氨酸、甘氨酸和胆碱代谢的脱氢酶。BN-PAGE揭示了响应于乙醇、SAM或两者的19种OxPhos蛋白水平的新变化。蛋白质组中的乙醇和SAM依赖性改变与基因表达的相应变化无关。总之，乙醇和SAM处理导致肝脏线粒体蛋白质组的多种变化。SAM对酒精毒性的保护作用部分是通过维持参与关键线粒体能量保存和生物合成途径的蛋白质来介导的。这项研究表明，SAM可能是治疗酒精性肝病的一个有前途的候选药物。
Andringa KK, King AL, Eccleston HB, Mantena SK, Landar A, Jhala NC, Dickinson DA, Squadrito GL, Bailey SM. Analysis of the liver mitochondrial proteome in response to ethanol and S-adenosylmethionine treatments: novel molecular targets of disease and hepatoprotection. Am J Physiol Gastrointest Liver Physiol 298: G732-G745, 2010. First published February 11, 2010; doi:10.1152/ajpgi.00332.2009.-S-adenosylmethionine (SAM) minimizes alcohol hepatotoxicity; however, the molecular mechanisms responsible for SAM hepatoprotection remain unknown. Herein, we use proteomics to determine whether the hepato-protective action of SAM against early-stage alcoholic liver disease is linked to alterations in the mitochondrial proteome. For this, male rats were fed control or ethanol-containing liquid diets +/- SAM and liver mitochondria were prepared for proteomic analysis. Two-dimensional isoelectric focusing (2D IEF/SDS-PAGE) and blue native gel electrophoresis (BN-PAGE) were used to determine changes in matrix and oxidative phosphorylation (OxPhos) proteins, respectively. SAM co-administration minimized alcohol-dependent inflammation and preserved mitochondrial respiration. SAM supplementation preserved liver SAM levels in ethanol-fed rats; however, mitochondrial SAM levels were increased by ethanol and SAM treatments. With use of 2D IEF/SDS-PAGE, 30 proteins showed significant changes in abundance in response to ethanol, SAM, or both. Classes of proteins affected by ethanol and SAM treatments were chaperones, beta oxidation proteins, sulfur metabolism proteins, and dehydrogenase enzymes involved in methionine, glycine, and choline metabolism. BN-PAGE revealed novel changes in the levels of 19 OxPhos proteins in response to ethanol, SAM, or both. Ethanol-and SAM-dependent alterations in the proteome were not linked to corresponding changes in gene expression. In conclusion, ethanol and SAM treatment led to multiple changes in the liver mitochondrial proteome. The protective effects of SAM against alcohol toxicity are mediated, in part, through maintenance of proteins involved in key mitochondrial energy conserving and biosynthetic pathways. This study demonstrates that SAM may be a promising candidate for treatment of alcoholic liver disease.