正常与一次性力竭运动小鼠骨骼肌基因表达谱差异的基因芯片检测

Microarray detection of gene expression profiles difference of skeletal muscle between normal rats and those with single bout of exhaustive exercise

目的:利用基因芯片技术了解小鼠骨骼肌基因表达谱,以及一次性力竭运动后小鼠骨骼肌基因表达谱的变化。方法:实验于2005-04/2005-06在卫生部北京老年医学研究所完成。①实验小鼠6只分为两组,正常对照组和一次性力竭运动组各3只。②从实验小鼠的骨骼肌组织中抽提总RNA;纯化后合成双链cDNA;体外转录合成生物素标记的cRNA;cRNA纯化后片段化为300bp左右的探针片断。③生物素标记的cRNA探针与涵盖了14000个明晰基因的Affymetrix基因表达谱芯片MouseGenome430A2.0杂交,分别检测正常对照组小鼠和一次性力竭运动组小鼠的骨骼肌基因表达谱。④利用生物信息学方法对检测结果进行分析。结果:6只实验小鼠进入结果分析。分别得到了正常对照组和一次性力竭运动组小鼠骨骼肌基因的表达谱,一次性力竭运动组与正常对照组相比,共筛选出差异表达基因189条,78条基因表达增加,111条基因表达降低。结论:利用基因芯片技术在阐明了小鼠骨骼肌基因表达谱的同时,提示运动性疲劳的产生涉及多个方面,为进一步研究运动性疲劳产生的机制提供了依据。

AIM：To explore the gene expression profiles of skeletal muscle in rat by microarray and the gene expression profiles alteration after single bout of exhaustive exercise. METHODS： The experiment was carried out in Beijiug-Institute of Geriatrics, Ministry of Health from April to June 2005. ①Totally 6 experimental rats were divided into 2 groups （ the group of normal control and the group of exhaustive exercised ）, with 3 in each group,② The total RNA was extracted from skeletal muscles of the experimental rats; Double-stand （ds-cDNA） was synthesized after the total RNA was purified; then cRNA labeled by biotin were synthesized by in vitro transcription; purified cRNA fragments were used as probe with the length of 300 bp.③ The skeletal muscle gene expression profiles of rats in 2 groups were tested by cRNA probes hybridizing with Affymetrix Mouse Genome 430A 2.0 gene chip, which approximately included 14 000 well-characterized genes. ④ The detected results were analyzed by bioinfonnatics method. RESULTS： Totally 6 rats were involved in the result analysis. The gene expression profiles of rats in 2 groups were obtained.Among the total 189 differential expression genes, selected by comparison, the expressions of 78 genes were up,regulated while 111 genes were down-regulated in exhaustive exercise rats compared with controls. CONCLUSION： The gene expression profile of skeletal muscle in rat is explored by microarray. Besides, it is indicated that the occurrence of exercise-induced fatigue results from the cooperation of many factors, providing data for the further research of the mechanism of exerciseinduced fatigue.