大鼠颈7神经根切断后肱三头肌蛋白质组学研究  

Proteomies analysis of triceps rnusele after C7 nerve root rhizotomy in rats

作　　者：黄亚福[1] 徐雷[1] 侯辰瑞 徐建光[1] 

机构地区：[1]复旦大学附属华山医院手外科,上海200040 [2]上海生命科学院生化细胞所蛋白质组研究分析中心

出　　处：《中华手外科杂志》2012年第3期169-172,共4页Chinese Journal of Hand Surgery

基　　金：国家自然科学基金资助项目（30872628）

摘　　要：目的用荧光差异双向凝胶电泳技术（2-D DIGE）分析大鼠颈7（C7）神经根切断后肱三头肌蛋白质表达变化规律，初步探讨其功能代偿的分子机制。方法建立一侧C7神经根切断大鼠模型，对侧为对照组，分别在C7神经根切断后第1、2、3、6、12周取等量的肱三头肌，用荧光差异双向凝胶电泳技术分离、单因素方差分析（one-way ANOVA）筛选蛋白质表达量差异具有统计学意义的蛋白质点（P〈0．05），再对差异蛋白质点进行胶内酶解，并用基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）或串联质谱技术进行鉴定。结果在大鼠岛神经根切断前和切断后的不同时间点，肱三头肌有38个蛋白质点表达发生了明显变化，其中有27个蛋白质被确切地鉴定，有11个蛋白质与肌纤维的能量代谢相关。在C7切断后第1周，无氧代谢酶表达下调，第3周无氧代谢酶、氧化酶（有氧代谢酶）表达均上调，但在第6～12周发生代谢类型的转变，无氧酶上调、氧化酶下调，同时线粒体蛋白质翻译延长因子表达下调。结论C7神经根切断后肱三头肌功能代偿机制可能与肌纤维类型的转变有关，而这种转变的调节可能发生在能量代谢酶的翻译水平。Objective To investigate the protein expression changes in rat triceps muscle after Ca nerve root rhizotomy by two way fluorescence differential gel electmphoresis （2D-DIGE）, and to study the molecular mechanism of the triceps muscle＇ s functional compensation. Methods C7 nerve root rkizotomy model was created on one side of the SD rats. The contralateral side did not undergo surgery and served as control. Equal amounts of triceps muscle were harvested from both sides after 1, 2, 3, 6 and 12 weeks. The total proteins from the C7 rhimtomy side and from the normal side were examined by the 2D-DIGE technique. One way ANOVA was used to screen protein spots that differentiated with statistical significance. These protein spots were then identified by MALDI-TOF-MS or MS/MS. Results At different time points following C7 rhizotomy there were 38 protein spots detected with significant change. Among them 27 proteins were identified by MALDI-TOF-MS or MS/MS. 11 proteins belonged to functional groups associated with the energy metabolism of muscles. One week after rat Ca nerve root rhizotomy, 5 protein spots of anaerobic metabolism enzyme down-regulated, then displayed an upregulation in their relative expression levels from 3rd to 12th week postoperatively. However 6 protein spots of aerobic oxidation enzymes down-regulated together with mitochondrial elongation factor Tu from 6th to 12th week. Conduslon The functional compensation of triceps after rat C7 nerve root rhizotomy may be associated with the changes in muscle fiber types, and the regulation of such a change may occur in die level of protein transhtion of energy metabolism enzyme.

关 键 词：蛋白质组学 周围神经 肱三头肌 荧光差异双向凝胶电泳技术 

分 类 号：R363[医药卫生—病理学]