烧冲复合伤早期大鼠肺组织蛋白质组学比较

Comparative proteomics study on lung tissue in early stage of burn-blast combined injury in rats

目的比较烧冲复合伤早期大鼠肺组织蛋白质组学表达差异。方法采用清洁级sD大鼠18只，用随机数表法分为烧冲复合伤组和单纯冲击伤组各9只。于致伤后6h，提取左侧肺组织蛋白。运用差异蛋白质组学方法，对两组肺组织蛋白提取物分别进行双向凝胶电泳（2-DE）。使用PDQuest图像分析，找出差异表达的蛋白，并进一步进行基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）分析，结合生物信息检索，确定烧冲复合伤早期大鼠急性肺损伤发生时差异表达的蛋白。同时观察大鼠伤后6h肺组织病理改变。结果去除实验中的死亡损耗后，最终每组均衡纳入8只大鼠进行统计学分析。2-DE凝胶结果显示，单纯冲击伤组肺组织得到736±47个蛋白点，烧冲复合伤组肺组织凝胶上共得到782±30个蛋白点，配比率为91％。PDQuest图像分析结果，有显著差异改变的蛋白点14个，其中烧冲复合伤组有10个点丰度显著增高（变化2倍以上），4个点丰度显著降低（变化2倍以上）。进一步行MALDI-TOF-MS分析，将得到的肽质量指纹峰谱（PMF）经过蛋白质数据库检索比对，得到12个有意义的蛋白质：参与应激反应的热休克蛋白27／70、参与代谢与免疫反应的碳酸甘酶2、细胞色素C氧化酶、ATP合成酶、Ca^2＋转运酶、参与细胞骨架的肌动蛋白结合蛋白2蛋白等。结论烧冲复合伤早期肺组织即出现了显著的蛋白质组学差异表达，其机制可能涉及与氧化应激、能量代谢变化、免疫应答反应、细胞骨架变化等相关蛋白。

Objective To explore changes in the proteomics of lung tissue in early stage of burnblast combined injury in rats. Methods According to a random digital table, a total number of 18 Sprague Dawley （ SD ） rats were divided into burn-blast combined injury group （ N = 9） and blast injury group （ n = 9）. Lung protein samples were collected at 6 h post injury. 2-DE was performed to separate proteins. After silver staining, the protein of differential expression were analyzed by PQ Quest and then identified by matrix-assisted laser desorption/ionization time of fight mass spectrometry （MALDI-TOF-MS）. The features of the changes of proteomics of rat lung after bum-blast combined injury were studied by biological spectrometry, protein bank and reference article analysis technique. At same time, pathologic changes of the lung were monitored after injury. Results After removing death drain during the experiment, each group contained 8 rats and the results were analyzed statistically. Well focused and distinct 2-DE maps with good reproducibility were obtained, means of 736 ± 47 and 782 ± 30 protein spots were detected from the blast injury group and burn-blast combined injury group and the matching rates were 91%. From the two groups, 14 differential protein spots expressions were analyzed by MALDI-TOF-MS, of which 10 proteins were up- regulated and 4 proteins were down-regulated in burn-blast combined injury group. 12 different expression proteins were identified in the lung through 2-DE, mass spectrometry and protein date base, including heat shock 27 protein 1, heat shock 70 protein 1, carbonic anhydrase 2, cytochrome c oxidase, ATP synthase subunit alpha, Ca ^2 ＋-transporting ATPase etal, which took part in stress reaction, metabolism, immune response and cytoskeleton. Conclusions Burn-blast combined injury could induce dramatically changes of proteomics in lung tissue at early stage. The mechanism probably involves several proteins associated with oxidative stress, energy metabolism, immune response and cytoskeleton.