Shanghai Public Health Clinical Center affiliated to Fudan University, Shanghai 201508, China.
Institute of Clinical Pharmacology, Pharmacogenetics Research Institute, Changsha, Hunan 410078, China.
Proteome Sci. 2012 Jun 8;10(1):39. doi: 10.1186/1477-5956-10-39.
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.
酒精性损伤已成为肝硬化的主要原因之一。然而，乙醇诱导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血浆膜蛋白质组学研究。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从肝纤维化 2 期和 4 期（S2 和 S4）的肝组织中纯化血浆膜。Western blot 验证其纯度。通过二维电泳（2DE）分离血浆膜蛋白，串联质谱鉴定差异表达蛋白。通过质谱鉴定出 S2 到 S4 之间有 16 个一致的差异表达蛋白。Western blot 验证差异表达蛋白 annexin A6 和 annexin A3 的表达，annexin A3 进一步通过免疫组化验证。我们的研究提示了乙醇改变蛋白表达以增强肝纤维化进展的可能机制。这些差异表达蛋白可能是治疗酒精性肝硬化的新药靶点。
