大鼠在体肺缺血再灌注损伤热休克蛋白25和硒结合蛋白1表达的关系

目的 应用蛋白质组学方法比较大鼠肺缺血再灌注(I/R)肺组织与正常对照组肺组织的差异蛋白质表达,探讨移植肺缺血再灌注损伤机制.方法 建立模拟在体左肺原位移植的大鼠改良模型,在缺血再灌注后4 h获取肺组织标本作为实验组(I/R组).蛋白质组技术比较I/R组和正常对照组大鼠肺组织的蛋白质变化,通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)分析获得肽质量指纹图谱,经Matrix Science查询软件搜索获得匹配的蛋白质.结果 获得分辨率较高、重复性较好的I/R组与对照组的双向凝胶电泳图谱,两组各自3块凝胶的平均蛋白质点数分别为489±52和511±83(P＞0.05),匹配率达89.28%和91.22%(P＞0.05).发现43个差异蛋白质点(P＜0.05),质谱分析鉴定出14个与肺缺血再灌注损伤相关的蛋白质.Western印迹分析证实HspSP25和硒结合蛋白l(SBP-I)的蛋白表达在I/R组早期显著升高(均P＜0.05).结论 肺缺血再灌注损伤后早期,具有应激保护功能的Hsp25和SBP-1等表达上调;而其他鉴定出的差异表达蛋白质口可能在能量代谢、组织抗应激、细胞凋亡及信号转导等方面发挥重要作用.

Relationship between the expression of heat shock protein 25 and selenium bidding protein 1 with ischemia/reperfusion lung injury in vivo: experiment with rats

Objective 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.Methotis 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 ditierential 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 veilfy part of the differentially expressed proteins.Resuits Well-resolved and reproducible 2-DE profile of rat L/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 all 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.Conclusion The protein expression of HSP25 and SBP-1 with stress protection function is up-regulated 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.