体神经-内脏神经吻合后异类神经再生过程的蛋白质组学分析

Proteomics analysis during foreign nerve regeneration after somatic-autonomic nerve anastomosis

目的使用蛋白质组研究技术研究大鼠体神经-内脏神经吻合术后异类神经再生过程中不同时段蛋白质表达的变化。方法建立大鼠体神经-内脏神经吻合动物模型,分别于术后7d、14d、28d取出再生的异类神经与对侧正常的体神经,应用双向电泳技术对提取物进行分析,经图像分析系统识别差异蛋白质,用MALDI-TOF-MS对差异表达的蛋白质进行鉴定分析,获得的肽质指纹图谱进入数据库中检索。结果质谱分析识别了25种具不同功能的蛋白质,其中包括神经髓鞘中的成分、急性期炎性因子、神经细胞骨架蛋白和脂类代谢蛋白等。这些蛋白可能在Wallerian变性、轴突再生和再髓鞘化中起作用。结论获得了再生异类神经时序性双向电泳凝胶图谱,成功鉴定25个与异类神经再生相关的蛋白质,揭示了施万细胞和巨噬细胞在异类神经再生过程中所起的重要作用。

Objective Using proteomics to explore the phasic expression profiles of proteins in rat during foreign nerve regeneration after somatic-autonomic nerve anastomosis. Methods The artificial somatic-autonomic nerve anatomosis were established in rats. Foreign nerves and contralateral somatic nerves were collected at 7,14 and 28 days respectively after operation. Extracts of foreign nerves were analyzed by two-dimensional gel eletrophoresis and the differential proteins were measured by matrix asisisted laser desorption ionization time of flight mass spectrometry. The data obtained from peptide mass fingerprinting （PMF） were used in protein database search and the protein identification. Results Mass spectrometric analysis identified 25 proteins displayed different function,including proteins in myeline sheath,inflammatory factors at acute stage,nerve cell cytoskeletal proteins and lipid metabolism proteins etc. The differential proteins may play a part at the process of Wallerian degeneration,axon regeneration and remyelinization.Conclusion Through proteome study we acquired time sequence protein expressions of regenerated foreign nerve,and successfully identified 25 proteins correlated with the process of foreign nerve regeneration. These results suggest that Schwann cells and macrophage cells may play important roles in the process of the foreign nerve regeneration.