bFGF长循环脂质体对大鼠脊髓牵张性损伤影响的蛋白质组学研究

PROTEOMICS STUDY ON EFFECT OF BASIC FIBROBLAST GROWTH FACTOR LONG CIRCULATION LIPOSOME ON SPINAL CORD TRACTION INJURY IN RATS

目的从蛋白质组学方面探讨bFGF长循环脂质体对大鼠脊髓牵张性损伤神经保护作用的可能机制。方法取20只SD大鼠随机分为脊髓牵张性损伤模型组(A组)和bFGF长循环脂质体治疗组(B组),每组10只。术后3周取材行脊髓组织蛋白的提取制备及定量;应用双向凝胶电泳对两组脊髓组织样品进行分离,建立差异表达图谱,通过凝胶成像分析和人工比对,分别确定差异蛋白点,采用超高效纳升液相色谱-电喷雾串联质谱(nano ultra-high performance liquid chromatography-electrospray tandem mass spectrometry,NanoUPLC-ESI-MS/MS)技术鉴定差异蛋白,检索数据库,对差异蛋白分类。结果两组间找到完全相同的差异蛋白点,B组与A组相比,得到差异蛋白点10个,其中6个下调,4个上调。采用NanoUPLC-ESI-MS/MS技术鉴定了26个差异蛋白点,共鉴定出18种有意义的蛋白;其中与凋亡相关的蛋白4种,神经传导和信号转导相关的蛋白3种,参与代谢的蛋白7种,1种功能不明,未知蛋白3种。结论bFGF长循环脂质体影响了大鼠牵张性脊髓损伤组织的蛋白质组表达,其对损伤脊髓的保护及修复作用可能是通过神经信号转导、调节细胞凋亡及细胞代谢等途径实现。

Objective To explore the possible active mechanism of the basic fibroblast growth factor （bFGF） long circulation liposome （LCL） （bFGF ＋ LCL） on spinal cord traction injury in rats at the level of proteomics. Methods Twenty Sprague Dawly rats were randomly divided into groups A and B, 10 rats in each group. The models of spinal cord traction injury was established at T12-L3 spines. The rats were not treated in group A, and the rats were treated with bFGF ＋ LCL （20 μg/ kg） in group B. At 3 weeks after operation, the rats were sacrificed for harvesting T13-L2 spinal tissue specimens. The protein was extracted and quantified in the spinal tissue firstly. The proteins from spinal tissue were separated by two-dimensional gel electrophoresis and identified by mass spectrometry. The different expression profiling was established in each group, and the differentially expressed protein was determined by comparing the level of each spot with gel imaging software and manually.The proteins were identified by nano ultra-high performance liquid chromatography-electrospray tandem mass spectrometry （NanoUPLC-ESI-MS/MS）, and the proteins were classified. Results The differentially expressed protein spots were found in 2 groups. Compared with group A, 4 spots were up-regulated and 6 were down-regulated in group B. NanoUPLC-ESI-MS/MS results showed that 18 significant proteins were identified in 26 differentially expressed proteins, including 4 apoptosis-related proteins, 3 nerve signal transduction related proteins, 7 proteins involved in metabolism, 1 unknown function protein, and 3 unnamed proteins. Conclusion The differentially expressed proteins are found in spinal cord traction injury of rats treated with bFGF ＋ LCL. bFGF ＋ LCL can affect the proteins expression in rats with spinal cord traction injury. The possible active mechanism is that it has protective and repair effects on injured spinal cord by nerve signal transduction, and regulation of nerve cells apoptosis and metabolism.