基于相对和绝对定量核素标记技术的大鼠海马线粒体蛋白质组学研究

Proteomics of the hippocampal mitochondria in rats based on isobaric tags for relative and absolutequantification technology

目的筛查深低温停循环时大鼠海马线粒体的差异蛋白质。方法以16G导管和22G留置针分别穿刺颈内静脉和尾动脉建立大鼠停循环10min模型，提取海马组织并纯化线粒体，超微结构验证，采用核素标记相对和绝对定量技术联合液相色谱-串联质谱技术获取肽段信息，Mascot2．2软件查库鉴定和分析获取差异蛋白质。结果共筛查到差异蛋白质8个（表达量2倍以上且P〈0．05），包括囊泡分拣蛋白45、无活性的磷脂酶c、假想蛋白loe365985、猫眼综合征染色体蛋白和Tid．1长亚型5个已知蛋白质，另外3个为未知蛋白质。结论通过8个线粒体差异蛋白质的变化发挥深低温停循环的脑保护作用。

Objective To screen the differentiall expressed proteins in the mitochondria of hippo campus in rats following deep hypothermic circulation arrest （DHCA）. Methods The model of circulation arrest with 10 min was created by puncture through the jugular vein and caudal artery by 16 G canal and 22 G detained needle, the hippocampus was extracted, and then hippocampal mitochondria were separated and validated at ultra-structure by electro-microscope. Peptide information was obtained by the methods of isobaric tags for relative and absolute quantification and liquid chromatography-mass spectrometry （MS）. The differentially expressed proteins were identified and analyzed by the soft of Mascot 2. 2. Results Eight proteins were revealed based on the standard that the relative ratio was larger than 2 or smaller than 0. 5 and P value less than 0. 05, including vacuolar protein sorting-associated protein 45, inactive phospholipase C-like protein 1, hypothetical protein LOC365985, cat eye syndrome chromosome region, Tid-1 long iso form and other three indefinite proteins. Conclusion Eight successfully identified proteins produce proba bly effect and Drotection after DHCA in hiDnocamnal mitochondria.