Liu Jian-xin, Wang Dian-jun, Zhang Qing-chun, Yin Bang-liang, Yang Jin-fu, Hu Jian-guo
Department of Thoracic and Cardiovascular Surgery, Third Hospital of Xiangya, Center Southern University, Changsha 410013, China.
Zhonghua Yi Xue Za Zhi. 2008 Jul 15;88(27):1929-32.
To analyze the differential expression proteins of rat ischemia/reperfusion (I/R) lung tissues in vivo and normal lung tissues by comparative proteome analysis, and to study the mechanism of donor lung I/R injury.
Forty male SD rats were randomly divided into 2 equal groups: I/R group undergoing mimic orthotopic left lung auto-grafting and harvesting of the left lung five hours after the operation, and control group undergoing isolation of the left hilus of lung and then harvesting of the left lung. The differential proteins in the left ventricle of transplanted heart were separated by means of immobilized pH gradient-based two-dimensional gel electrophoresis (2-DE), identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), and searched through Matrix Science software system. Western blotting was used to verify part of the differentially expressed proteins.
Well-resolved and reproducible 2-DE profile of rat I/R lung tissues and normal lung tissues were obtained. In the I/R lung tissue profile, the average spot number from 3 gels was 489 +/- 52 spots (P > 0.05) with an average matching rate of 89.28% (P > 0.05), and in the control group, the average spot number from 3 gels was 511 +/- 83 spots (P > 0.05) with an average matching rate of 91.22% (P > 0.05). Fourteen differential proteins were identified by peptide mass fingerprinting (PMF) searched in Matrix Science (P < 0.05). Western blotting confirmed that the protein expression of selenium binding protein 1 (SBP-1) and heat shock protein 25 (HSP25) increased at the early stage of I/R group.
The protein expression of HSP25 and SBP-1 with stress protection function is upregulated in the early stage of lung I/R injury. Other differentially expressed proteins identified may have important functions in energy metabolism, tissue stress, cell apoptosis, and signal transduction.
通过比较蛋白质组分析，分析大鼠体内缺血/再灌注（I/R）肺组织与正常肺组织中差异表达的蛋白质，探讨供体肺I/R损伤的机制。
40只雄性SD大鼠随机分为2组：I/R组行模拟原位左肺自体移植，术后5小时取左肺；对照组行左肺门分离后取左肺。采用基于固定化pH梯度的双向凝胶电泳（2-DE）分离移植心脏左心室中的差异蛋白质，用基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF-MS）进行鉴定，并通过Matrix Science软件系统进行检索。采用蛋白质印迹法验证部分差异表达蛋白质。
获得了分辨率良好且可重复的大鼠I/R肺组织和正常肺组织的2-DE图谱。在I/R肺组织图谱中，3块凝胶的平均点数为489±52个点（P>0.05），平均匹配率为89.28%（P>0.05）；对照组3块凝胶的平均点数为511±83个点（P>0.05），平均匹配率为91.22%（P>0.05）。通过在Matrix Science中搜索肽质量指纹图谱（PMF）鉴定出14种差异蛋白质（P<0.05）。蛋白质印迹法证实，I/R组早期硒结合蛋白1（SBP-1）和热休克蛋白25（HSP25）的蛋白表达增加。
具有应激保护功能的HSP25和SBP-1蛋白在肺I/R损伤早期表达上调。鉴定出的其他差异表达蛋白质可能在能量代谢、组织应激、细胞凋亡和信号转导中具有重要作用。
