大鼠感觉、运动神经瓦勒氏变性前后蛋白分析

Protein expression profiling during wallerian degeneration in sensory and motor nerve in rat

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 Isobaric tags for relative and absolute quantitation（iTRAQ） and online two-dimensional liquid chromatographytandem mass spectrometry（2D LC-MS/MS） were performed to identify differentially expressed proteins in sensory and motor nerves at 7 days after transaction. Methods Forty adult male wistar rats weighing 220-250 g were randomly divided into control group and experimental group（n=20 each group）. Spinal cord transaction was performed to rats in experimental group. Seven days later, samples of normal/injured ventral root nerve and dorsal root nerve were obtained and labeled using iTRAQ（114, 115, 116, 117 were used for labeling normal motor nerve, injured motor nerve, normal sensory nerve, injured motor nerve respectively）, and analyzed by 2DLC-MS/MS. Results A total of 4 312 proteins were identified. 1）There were 637 differentially expressed proteins in the injured motor nerve compared to the normal motor nerve, in which 265 were up-regulated and 372 were down-regulated. 2）There were 626 differentially expressed proteins in the injured sensory nerve compared to the normal sensory nerve, in which 258 were upregulated and 368 were down-regulated proteins. Conclusion We screen out differentially expressed proteins in sensory and motor nerve in the process of Wallerian degeneration, and this may contribute to the research in peripheral nerve regeneration.