摘要
目的:研究缺血再灌注(ischemia/reperfusion,I/R)对大鼠骨骼肌组织蛋白质表达谱的影响。方法:健康雄性Wistar大鼠12只随机分为2组(n=6):假手术组和I/R组,无创动脉夹夹闭右侧股动脉4h,松夹再灌注24h建立I/R模型;实验结束时提取骨骼肌组织蛋白质,双向电泳技术分离骨骼肌组织蛋白质,分析差异显示的蛋白质并选取7个差异显著的蛋白点进行质谱分析。结果:双向电泳可分离(354±13)个蛋白质,点匹配率为(78.7±1.4)%,I/R诱导骨骼肌组织10种蛋白质出现明显差异表达,其中6种表达上调,3种表达下调,1种在I/R组为2个点。质谱鉴定出5个蛋白质为:线粒体醛脱氢酶(mitochondrial aldehyde dehydrogenase,ALDH)前体、热休克蛋白27(heat shock 27kD protein,HSP27)和一未命名蛋白(I/R组表达升高)、α-肌动蛋白(α-actin,I/R组表达下降)、核转移因子2(nuclear transport factor2,NTF-2)在I/R组发生突变。结论:I/R损伤引起大鼠骨骼肌蛋白质表达发生改变,其中α-肌动蛋白、ALDH和HSP27表达及NTF-2突变可能与I/R损伤有关。
AIM： To investigate ischemia reperfusion （I/R） -induced proteomic changes in rat skeletal muscle. METHODS： Healthy male Wistar rats were randomly divided into two groups as follows （n = 6） ： sham group and I/R group. I/R of fight hind limb was induced by 4 h ischemia followed by 24 h reperfusion. The 2 - DE was applied to separate the proteins extracted from skeletal muscle tissue at the end of experiment, followed by Coomassie Brillant blue R- 250 staining. Computer image analysis was used to determine the differential expression of proteins between the two groups, and 7 protein spots expressed differentially were picked out and subjected to in - gel digest and MALDI - TOP for identification. RESULTS： 354±13 proteins were detected and the match rate was （78.7±1.4）%. 10 proteins displayed significant changes after I/R, of which, 6 proteins increased and 3 proteins decreased in expression. Moreover, 2 spots in I/R group were observed, only 1 spots of which in control. 5 proteins were identified after mass spectrometry. Mitochondrial aldehyde dehydrogenase （ALDH） precursor, heat shock 27 kD protein （HSP27）, an unnamed protein product （increased in I/R group）, α -actin （decreased in I/R group）, and nuclear transport factor 2 （NTF -2） W7a mutant were found in I/R group. CONCLUSION： I/R injury induced differential proteomic changes in rat skeletal muscle. ALDH, α -actin and HSP27 expression, and NTF- 2 mutation are involved in I/R injury.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2008年第11期2186-2190,共5页
Chinese Journal of Pathophysiology
基金
国家自然科学基金资助项目(No.30670822
No.30770902)
国家自然科学基金重大国际合作资助项目(No.30620130111)
国家重点基础研究发展计划资助项目(No.2007CB512003)
关键词
再灌注损伤
蛋白质组
电泳
双向
凝胶
肌
骨骼
Reperfusion injury
Proteome
Electrophoresis, two-dimensional,gel
Muscle, skeletal
