抗阻训练改善骨骼肌衰老的关键基因与信号通路鉴定

Identification of Crucial Differentially Expressed Genes and Signaling Pathways in Ameliorating Skeletal Muscle Aging by Resistance Training

目的:探讨抗阻训练对衰老骨骼肌基因表达谱的影响及其生物代谢过程变化,筛选出骨骼肌衰老干预的关键基因与信号通路。方法:从GEO数据库筛选下载3套衰老骨骼肌基因表达谱数据(GSE 25941、GSE 38718与GSE 8479)作为分析材料,进行标准化后,利用R软件筛选3套数据集中的差异表达基因。重叠分析法整合筛选共同显著的差异基因,采用DAVID、KOBAS和Revigo数据库对其基因本体论(Gene Ontology,GO)和京都基因与基因组百科全书代谢通路数据库(Kyoto EncyclopediaofGenesandGenomes,KEGG)的注释进行功能富集分析。STRING数据库对差异基因编码蛋白进行蛋白质-蛋白质互作分析,筛选出关键基因,进行二次功能富集分析,以明确关键通路。结果:通过整合分析,筛选出抗阻训练对衰老骨骼肌影响的差异表达基因共计60个。差异基因编码蛋白间的相互作用主要集中于MXRA5、JAK1、CCNG1、FOS、MYH1、RHOQ、ZAK、ASB11、CCNI、KIF5B、DUSP1、MYL5、ASB15、PRKAA2与SLMAP共计15个关键基因。其中,ZAK、DUSP1、FOS、JAK1、RHOQ和PRKAA2主要富集于MAPK、胰岛素与PI3K-Akt3条KEGG代谢通路,基因本体涉及骨骼肌响应刺激、细胞内信号转导、运动(神经元)活性、核苷酸及蛋白质结合等生物过程与分子功能。结论:ZAK、DUSP1、FOS、JAK1、RHOQ和PRKAA2可能是临床干预骨骼肌衰老的关键靶点,通过MAPK、胰岛素、PI3K-Akt甚至AMPK信号通路介导响应抗阻训练,调节下游靶蛋白功能表达,最终改善衰老骨骼肌表型。

Objective:To investigate the effects of resistance exercise training on gene expression profile and biological metabolism,and screen out candidate targets involved in clinical intervention against skeletal muscle aging.Methods:Three microarray datasets(GSE25941、GSE38718 and GSE8479)of aging skeletal muscle were downloaded from GEO database.After the normalization of raw data,differentially expressed genes(DEGs)were screened by using Limma package within R software.The overlapping analysis was then used to screen out the co significant DEGs in three datasets,and the study applied enrichment analysis including Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway on these DEGS with DAVID,KOBAS and Revigo databases.Finally,the study performed the protein protein interactive analysis based on DEGs encoded proteins to identify the candidate hub genes,and performed the enrichment analysis again to identify the crucial pathways which seemed to be involved in the responses of aging skeletal muscle to resistance exercise training.Results:A total of 60 DEGs were screened out as the candidate molecular signature may be responsible for the effects of resistance exercise training on aging skeletal muscle.And,15 DEGs encoded proteins including MXRA5,JAK1,CCNG1,FOS,MYH1,RHOQ,ZAK,ASB11,CCNI,KIF5B,DUSP1,MYL5,ASB15,PRKAA2 and SLMAP were found to be the most active proteins.Furthermore,6 genes i.e.ZAK,DUSP1,FOS,JAK1,RHOQ and PRKAA2 were found to be enriched in MAPK,insulin,and PI3K-Akt signaling pathway,while the biological processes and molecular functions were found to be enriched in response to stimuli,intracellular signaling,motor activity,ribonucleotide and protein binding.Conclusions:ZAK,DUSP1,FOS,JAK1,RHOQ and PRKAA2 might function as the candidate targets involved in the ameliorating effects of resistance exercise training on skeletal muscle aging.While MAPK,insulin,PI3K-Akt and even AMPK signaling pathway might play the central role in mediating the adaptive response to resistance training stimuli.