论文部分内容阅读
目的应用蛋白质双向凝胶电泳(two-dimensional polyacrylamide gel electrophoresis,2-DE)技术分析在急性重度失血性休克(acute severe hemorrhagic shock,ASHS)条件下大鼠血浆蛋白质组表达的差异。方法16只雄性Wistar大鼠随机分成对照组(sham hemorrhage shock,SHS)和ASHS模型组,每组8只。采用股动脉放血的方法制备模型,在规定时间内处死大鼠并收集血浆,去除血浆中的高丰度蛋白后进行2-DE,运用Image Master 2D Platinum v 5.0凝胶图像分析软件对2-DE凝胶图像进行差异表达分析,有意义的差异蛋白质点用基质辅助激光解析电离飞行时间质谱进行肽质量指纹图谱分析,借助Swiss-prot数据库进行蛋白质搜索和鉴定。结果SHS组和ASHS组肝脏的2-DE图谱分别平均识别到(570.0±0.9)和(578.0±1.4)个蛋白质点,SHS组和ASHS组肝脏间平均匹配率达83.6%~86.2%,共发现8个差异有意义的蛋白质点,鉴定出了白蛋白、甲状腺激素结合蛋白-D链蛋白和信号素-3D前体等3种蛋白质。结论以2-DE技术得到重复性和分辨率都较好的2-DE图谱,并初步鉴定急性重度失血性休克后大鼠血浆去除高丰度蛋白后的差异表达蛋白质,为深入研究失血性休克的生理病理机制及寻找失血性休克预防和治疗的生物标志物提供了依据。
Objective To analyze the difference of proteome expression in rats with acute severe hemorrhagic shock (ASHS) by two-dimensional polyacrylamide gel electrophoresis (2-DE). Methods Sixteen male Wistar rats were randomly divided into control group (sham hemorrhage shock, SHS) and ASHS model group (n = 8). The rats were sacrificed and the plasma was collected. The 2-DE protein was removed from the plasma and the 2-DE was imaged using Image Master 2D Platinum v 5.0 gel image analysis software. Gel images were differentially expressed and analyzed. Significantly different protein spots were analyzed by peptide mass fingerprinting with matrix-assisted laser desorption / ionization time of flight mass spectrometry, and protein searches and identification were performed using the Swiss-prot database. Results The 2-DE maps of the liver of SHS group and ASHS group were (570.0 ± 0.9) and (578.0 ± 1.4), respectively. The average matching rate of liver between SHS group and ASHS group was 83.6% ~ 86.2% Eight differentially significant protein spots identified three proteins, albumin, thyroid hormone-binding protein-D chain protein and signal-3D precursor. Conclusions The 2-DE pattern with good reproducibility and resolution was obtained by 2-DE technique. The differentially expressed proteins after high-abundance protein removal in plasma of rats with acute severe hemorrhagic shock were initially identified. In order to further study the hemorrhagic shock The physiological and pathological mechanisms and find a biomarker for the prevention and treatment of hemorrhagic shock.