铜绿假单胞菌相关性肺损伤的蛋白质差异表达研究  

Differential expression of proteins in Pseudomonas aeruginosa-induced acute lung injury

作　　者：刘冬冬[1] 毛璞[1] 廖东江[1] 余志辉 杨淳[1] 黄红川[1] 刘晓青[1] 何为群[1] 黎毅敏[1] 

机构地区：[1]广州医学院第一附属医院广州呼吸疾病研究所,510120

出　　处：《国际呼吸杂志》2011年第12期896-902,共7页International Journal of Respiration

基　　金：基金项目：广东省科技计划项目（20088060600048）;广州市教育局创新学术团队（B94117）

摘　　要：目的利用蛋白质组学方法筛选铜绿假单胞菌急性肺损伤特异相关蛋白质，为鉴别诊断感染与非感染因素引起的急性肺损伤提供理论依据。方法建立感染（铜绿似单胞菌急性肺损伤）与非感染（包括机械通气急性肺损伤、油酸急性肺损伤）大鼠模型，应用双向凝胶电泳技术分离总蛋白。Image Master双向凝胶电泳软件分析差异表达蛋白质点后，应用基质辅助激光解析电离飞行时间质谱获取肽质量指纹图谱并通过NCBI数据库鉴定差异表达的蛋白质，最后Western blot验证差异表达的蛋白质。结果双向凝胶电泳图谱中找到32个表达量有明显差异的蛋白质点，质谱鉴定出18种蛋白质。其中的过氧化物氧化还原酶I（PrxI）、钙网蛋白等11种蛋白质在铜绿假单胞菌急性肺损伤组表达上调，并在Westernblot验证了PrxI的上调，与凝胶电泳结果一致。谷胱甘肽-S-转移酶a4、甲状腺素蛋白等7种蛋白在铜绿假单胞菌急性肺损伤组表达下调。结论应用比较蛋白组学鉴定出18种差异蛋白，11种在铜绿假单胞菌急性肺损伤组表达上调，7种表达下调，分别参与催化代谢、氧化还原、信号转导等凋节。其中PrxI和钙网蛋白可能通过对氧化／抗氧化和免疫应答的调节在细菌感染性急性肺损伤中起着重要作用。本研究为进一步阐明感染相关性急性肺损伤的发病机制提供新的线索。Objective To screen the differential expressed proteins of Pseudomonas aeruginosainduced acute lung injury by proteomies,and to provide the theoretical basis for the identification of acute lung injury caused by bacterial infection and non infection. Methods The rat models of Pseudomonas aeruginosa-induced acute lung injury and non infection-induced acute lung injury （including ventilator induced acute lung injury and oleic acid-induced acute lung injury） were established. The total proteins were extracted and separated by two-dimensional gel electrophoresis （2 DE）. The sites of differential expression were analyzed using Image Master 2D Elite 5.0 image analysis software,and the peptide mass finger printing was identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF MS）. The biological information of these proteins was searched in the NCBI database and confirmed by Western blot. Results Thirty-two differential expression protein spots were found in 2-DE maps, and eighteen proteins were analyzed and identified by MAI.DI TOF MS. Eleven proteins （including peroxiredoxin I , calreticulin etc. ） were upregulated while seven proteins （including glutathione S-transferasea4, transthyretin etc. ） were downregulated in Pseudomonas aeruginosa-induced acute lung injury group. The upregulated expression of peroxiredoxin I in Pseudomonas aeruginosa induced acute lung injury group was confirmed by Western blot. Conclusions Eighteen different proteins were identified by comparative proteomics, and involved in catalytic metabolism, redox, signal transduction. Eleven proteins were upregulated and seven proteins were downregulated in Pseudomonas aeruginosa-induced acute lung injury group. Peroxiredoxin I and calreticulin may play an important role in bacterial infected-acute lung injury by regulating oxidant/antioxidant and immune response.

关 键 词：急性肺损伤 铜绿假单胞菌 双向凝胶电泳 

分 类 号：R563.8[医药卫生—呼吸系统]