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目的通过同位素标记相对和绝对定量(isobaric tags for relative and absolute quantitation,iTRAQ)技术联合二维液相色谱-串联质谱(liquid chromatography-tandem mass spectrometry,2D LC-MS/MS)技术研究感觉、运动神经损伤7 d后蛋白表达差异。方法成年雄性Wistar大鼠40只,体质量220~250 g,随机分为对照组和实验组,各20只。实验组大鼠脊髓横断,对照组不作处理。7 d后对两组大鼠脊神经前后根神经取材,运用iTRAQ技术标记(114标记正常运动神经;115标记损伤的运动神经;116标记正常的感觉神经;117标记代表损伤的感觉神经)并结合2D LC-MS/MS技术进行差异蛋白分析,并利用基因本体论和京都基因与基因组百科全书信号通路分析差异蛋白所在信号通路。结果共鉴定出4 312个蛋白:1)其中运动神经损伤后差异蛋白数量为637个,表达升高的有265个,表达降低的有372个;2)感觉神经损伤后差异蛋白数量为626个,表达升高的有258个,表达降低的有368个。结论成功筛选出大鼠运动神经与感觉神经损伤前后的差异蛋白,可为周围神经再生研究提供支持。
OBJECTIVE: To study the effects of sensory and motor nerves on the basis of isobaric tags for relative and absolute quantitation (iTRAQ) combined with two-dimensional liquid chromatography-tandem mass spectrometry (2D LC-MS / MS) The protein expression difference after 7 d injury. Methods Forty adult male Wistar rats weighing 220-250 g were randomly divided into control group and experimental group, with 20 rats in each group. Spinal cord transection in experimental group, no treatment in control group. After 7 days, the anterior and posterior root nerves of the two groups of rats were harvested and labeled with iTRAQ technique (114 marks normal motor nerves; 115 marks damaged motor nerves; 116 marks normal sensory nerves; 117 marks damaged sensory nerves) combined with 2D LC-MS / MS techniques were used to analyze differential proteins and to use the ontology and Kyoto Encyclopedia of Gene and Enzymes to analyze the signaling pathways involved in differentially expressed proteins. Results A total of 4 312 proteins were identified: 1) The number of differentially expressed proteins was 637 after motor nerve injury, 265 of them were elevated and 372 were decreased. 2) The number of differentially expressed proteins after sensory nerve injury was 626 258 were up-regulated and 368 were down-regulated. Conclusion The successful screening of differentially expressed proteins between motor and sensory nerve in rats may provide support for the study of peripheral nerve regeneration.