Klouckova Iveta, Hrncirova Petra, Mechref Yehia, Arnold Randy J, Li Ting-Kai, McBride William J, Novotny Milos V
Department of Chemistry, Indiana University, 800 E. Kirkwood Avenue, Bloomington, IN 47405, USA.
Proteomics. 2006 May;6(10):3060-74. doi: 10.1002/pmic.200500725.
This study compares the total liver proteome of inbred alcohol-preferring line (iP) rats exposed to alcohol with iP rats without alcohol experience. Rat liver proteins were extracted using a three-step procedure. Each of the three solutions solubilizes a different set of proteins. The extracted proteins were separated by 2-DE. Scanned gels of two sample groups, alcohol-exposed iP and alcohol-naïve iP, were compared, revealing many protein spots with significantly higher or lower densities. These spots were cut from the gel, destained, and subjected to trypsin digestion and subsequent identification by LC-MS/MS. Twenty-four individual rats, 12 alcohol-naïve, and 12 alcohol-exposed, were used in this study. Two groups, each containing six naïve and six exposed animals, were created for statistical comparison. For the first group, 64 spots were observed to have statistically significant intensity differences upon alcohol exposure across all three extracts while 118 such spots were found in the second group. There were 113 unique proteins in both groups together. The majority of these proteins were enzymes. Significant changes are observed for three major metabolic pathways: glycolysis, gluconeogenesis, and fatty acid beta-oxidation. In addition, enzymes involved in protein synthesis and antioxidant activity show significant changes in abundance in response to alcohol exposure.
本研究比较了有酒精接触史的近交系嗜酒大鼠（iP）与无酒精接触史的iP大鼠的肝脏蛋白质组。采用三步法提取大鼠肝脏蛋白质。三种溶液中的每一种都能溶解不同组别的蛋白质。提取的蛋白质通过双向电泳（2-DE）进行分离。比较了酒精接触组iP和未接触酒精组iP两个样本组的扫描凝胶，发现许多蛋白质斑点的密度显著更高或更低。从凝胶上切下这些斑点，脱色，然后进行胰蛋白酶消化，并随后通过液相色谱-串联质谱（LC-MS/MS）进行鉴定。本研究使用了24只大鼠，其中12只未接触酒精，12只接触酒精。为进行统计学比较，创建了两组，每组包含6只未接触酒精的动物和6只接触酒精的动物。对于第一组，在所有三种提取物中，观察到64个斑点在酒精接触后具有统计学上显著的强度差异，而在第二组中发现了118个这样的斑点。两组共有113种独特的蛋白质。这些蛋白质大多数是酶。在三个主要代谢途径中观察到了显著变化：糖酵解、糖异生和脂肪酸β氧化。此外，参与蛋白质合成和抗氧化活性的酶在酒精接触后丰度显示出显著变化。
