CAJ | 학술논문

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C7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.
목적 용형광차이쌍향응효전영기술(2-D DIGE)분석대서경7(C7)신경근절단후굉삼두기단백질표체변화규률,초보탐토기공능대상적분자궤제.방법 건립일측C7신경근절단대서모형,대측위대조조,분별재C7신경근절단후제1、2、3、6、12주취등량적굉삼두기,용형광차이쌍향응효전영기술분리、단인소방차분석(one-way ANOVA)사선단백질표체량차이구유통계학의의적단백질점(P＜0.05),재대차이단백질점진행효내매해,병용기질보조격광해흡전리비행시간질보(MALDI-TOF-MS)혹천련질보기술진행감정.결과 재대서C7신경근절단전화절단후적불동시간점,굉삼두기유38개단백질점표체발생료명현변화,기중유27개단백질피학절지감정,유11개단백질여기섬유적능량대사상관.재C7절단후제1주,무양대사매표체하조,제3주무양대사매、양화매(유양대사매)표체균상조,단재제6～ 12주발생대사류형적전변,무양매상조、양화매하조,동시선립체단백질번역연장인자표체하조.결론 C7신경근절단후굉삼두기공능대상궤제가능여기섬유류형적전변유관,이저충전변적조절가능발생재능량대사매적번역수평.
Objective To investigate the protein expression changes in rat triceps muscle after C7 nerve root rhizotomy by two way fluorescence differential gel electrophoresis (2D-DICE),and to study the molecular mechanism of the triceps muscle's functional compensation. Methods C7 nerve root rhizotomy 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 rhizotomy 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.11proteins belonged to functional groups associated with the energy metabolism of muscles.One week after rat C7 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.Conclusion 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 the level of protein translation ofenergy metabolism enzyme.