| 摘要: |
| 目的：应用蛋白质组学技术观察大鼠肝脏大部分切除术后蛋白表达情况，筛选肝脏切除后肝再生相关的差异表达蛋白。方法：大鼠随机分为肝大部切除组（n=35）和假手术对照组（n=5）。肝大部切除组大鼠无菌条件下切除肝左外叶、左中叶和中叶共约70%肝脏；假手术组大鼠仅开腹，不行肝大部切除。肝大部切除术后不同时间点（2、12、24、36、48、72、168 h）各处死5只大鼠，取右叶肝组织，提取大鼠肝脏总蛋白，以假手术组为对照，进行二维电泳和质谱分析，筛选差异表达蛋白，并对其中差异显著蛋白进行Western印迹鉴定。结果：电泳图谱显示肝脏70%切除术后2 h差异表达蛋白点开始增多，36 h达到高峰；共筛选出78个差异表达蛋白点，质谱技术鉴定出其中35个有意义的蛋白点。35个差异蛋白根据动态变化趋势可分为5类：3类丰度上调蛋白和2类丰度下调蛋白（各自间上调或下调时间和幅度均有所差异）；这些蛋白主要涉及以下代谢途径：氧化应激反应、急性期反应蛋白、脂类代谢蛋白、能量代谢的酶类、信号转导、神经递质降解等，部分蛋白功能未知。Western印迹鉴定证实其中的prohibitin蛋白在肝大部切除术后2 h开始上调，36 h达高峰，48 h后趋于正常水平。结论：多种信号和代谢途径参与肝脏大部切除后的肝脏再生过程。 |
| 关键词: 肝再生 肝切除术 蛋白质组学 prohibitin |
| DOI：10.3724/SP.J.1008.2008.00031 |
| 投稿时间：2007-10-09修订日期：2007-12-28 |
| 基金项目: |
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| Proteomics technique in screening of differential proteins for liver regeneration after partial hepatectomy in rats |
| ZHOU Dong-xun, TAN Ye-xiong, HU He-ping*, WANG Hong-yang*, WU Meng-chao |
| (Department of Hepatobiliary Disease Ⅱ, Eastern Hepatobiliary Hospital, Second Military Medical University, Shanghai 200438, China) |
| Abstract: |
| Objective：To screen for the differentially expressed proteins during rat liver regeneration after partial hepatectomy(PH) by proteomics technique. Methods: Healthy male SD rats were randomly divided into 2 groups: hepatectomy group and sham operated group. The hepatectomy model was produced by 70% PH(n=35) and the sham operated rats（n=5）underwent the same surgical protocol without hepatectomy. Rats were executed at 2, 12, 24, 36, 48, 72 and 168 h after partial hepatectomy (each time 5 rats) and the right lobes were harvested. The total protein was extracted and analyzed by two-dimensional gel electrophoresis and mass spectrometric analysis. The differential proteins were then analyzed by Western blotting. Results: The spots of differential protein began to rise at 2 h after PH and peaked at 36 h after PH. A total of 78 protein spots were identified and 35 significant protein spots were found by mass spectrometric analysis. The 35 protein sports fell into 5 types according to their dynamic changes: 3 up-regulated and 2 down-regulated (with different regulation time periods and amplitudes); and their functions involved oxidative stress response, acute reaction, lipid and energy metabolism, intracellular signaling transduction, cell proliferation, etc., with some having unknown functions. Western blotting analysis showed that the prohibitin protein began to increase 2 h after PH and decreased to the normal level after 48 h. Conclusion: It is indicated that many proteins and signal transduction pathways participate in the liver regeneration after partial hepatectomy. |
| Key words: liver regeneration hepatectomy proteome prohibitin |
