摘要
目的实施大鼠体外循环(CPB)前后血浆比较蛋白质组学研究,了解CPB前、后血浆蛋白质的差异性表达,以期发现对早期诊断CPB后并发症有参考价值的血浆标志物。方法将成年雄性SD大鼠10只分为实验组和对照组,每组5只,术前自由进食进水。实验组大鼠建立CPB模型,对照组只进行麻醉和动、静脉穿刺操作。两组动物分别在CPB前和CPB结束时采取血液标本1mL,肝素抗凝,分离血浆。血浆蛋白提纯后进行二维凝胶电泳、用ImageScanner扫描仪扫描成像。将被认定为差异表达的蛋白质斑点进行切胶、酶解和肽段提取后,用质谱仪分析鉴定。结果 CPB后,大鼠血浆蛋白质表达斑点数目明显增加,共有17个表达上调的蛋白质斑点被认定为CPB所致。经质谱分析,共鉴定出5种蛋白质,分别为:凝溶胶蛋白、结合珠蛋白、载脂蛋白A1(ApoA1)、免疫球蛋白和Ba1-647。结论 CPB可引起大鼠血浆蛋白质出现差异性表达,从功能上分析,凝溶胶蛋白、结合珠蛋白、ApoA1有可能作为研究CPB并发症的血浆标志物。
Objective To understand the plasma protein differential expression before and after cardiopulmonary bypass（CPB）through conducting the comparative proteomics study on rats in order to find the plasma markers with potential value in the early diagnosis of CPB resulted complications.Methods 10 adult male SD rats were divided into the experiment group and the control group randomly（n=5）,and took food and water freely before operation.The rat models of CPB were constructed in the experiment group.But no any CPB operation was administered in the control group in addition to anesthesia induction,arterial and venous puncture procedure.1mL of blood sample was extracted for separating plasma before CPB and at the end of CPB in the two groups.The total plasma protein was purified.Then the 2-dimensional electrophoresis and the scanning imaging by ImageScanner were performed.The protein spots verified to be differential expression were performed the cutting,enzymolysis and peptide fragment extraction.Finally the mass spectrometer was adopted to conduct the analysis and identification.Results The number of visualized spots was increased significantly after CPB.17 protein spots with up-regulated expression were identified as differential expression caused by CPB.5proteins were verified by mass spectrometer analysis and database research.They were gelsolin,haptoglobin,apolipoprotein A-1,immunoglobulin gamma-2band Ba1-647 respectively.Conclusion CPB can cause the differential expression of plasma proteins in rat model.According to the function analysis,gelsolin,haptoglobin and apolipoprotein A-1have the potentiality of being the plasma markers for studying CPB complications.
出处
《重庆医学》
CAS
北大核心
2015年第10期1311-1313,共3页
Chongqing medicine
基金
国家自然科学基金资助项目(30700785)
中国博士后科学基金资助项目(200704100779)
