衰老过程中骨骼肌microRNA-mRNA调控网络分析

doi:10.3969/j.issn.1003-9198.2022.04.008

摘要/Abstract

摘要： 目的整合高通量 miRNA/mRNA表达数据,探究肌肉衰老过程中微小RNA(miRNA)和mRNA的变化。方法利用2个不同年龄(6个月和24个月)小鼠腓肠肌的mRNA数据库,通过“差异miRNA 靶向调控基因”和差异mRNA联合分析,构建骨骼肌衰老过程中的核心调控网络。使用表达分析系统(GSEA)软件进行差异表达基因基因本体(GO)和京东基因与基因组百科全书(KEGG)信号通路分析。采用实时荧光定量聚合酶链式反应(qRT-PCR)和蛋白免疫印迹检测代表性差异miRNA和靶向mRNA表达。结果青年和老年小鼠骨骼肌差异表达的基因和miRNA分别为877个和84个。构建miRNA-mRNA网络,结果包含21个miRNA,其中靶向基因超过7个的miRNA有9个,qRT-PCR验证均与测序结果一致。通路分析显示,差异基因主要与转录、代谢过程和骨骼肌分化过程相关,骨骼肌分化过程中包含了早期生长反应蛋白2(EGR2), 环1(Ring1)人转录因子 Yin-Yang1(YY1)和 Ring1 和 YY1 结合蛋白(Rybp)基因。qRT-PCR验证骨骼肌分化过程中4个基因的mRNA水平均升高,Rybp在蛋白质水平上也有所增加。结论 miRNA可能通过参与转录、代谢和骨骼肌分化等过程来促进肌肉衰老。Rybp及其推测的调控性miRNA miR-136可能促进了肌肉衰老过程。

关键词: 肌肉衰老, 微小RNA, 基因, 骨骼肌

Abstract: Objective To explore the changes of miRNA and mRNA during muscle aging through integrating high-throughput microRNA(miRNA)/mRNA expression data. Methods Two different age(6 and 24 months) mice gastrocnemius miRNA databases were used, and the combined analysis of “differential miRNA targeted regulatory genes” and differential mRNA was carried out. The core regulatory network in the skeletal muscle aging process was constructed. GSEA software was used to analyze the gene ontology(GO) function and Kyoto encyclopedia of genes and genomes(KEGG )signaling pathway involved in the differential gene. Representative differential miRNA and targeted mRNA expression were detected by real-time fluorescent quantitative PCR (qRT-PCR)and Western blot. Results A total of 877 and 84 differentially expressed genes and miRNAs were identified in the skeletal muscle of young and old mice, respectively. The construction of a miRNA-mRNA network resulted in 21 miRNAs, and among them, there were 9 miRNAs with more than 7 target genes, and the qRT-PCR verification is consistent with the sequencing results. Pathway analysis showed that the differential genes were mainly related to transcription, metabolism and skeletal muscle differentiation. The skeletal muscle differentiation process contained early growth response protein 2(EGR2), Ring1, transcription factor Yin-Yang1(YY1)and Ring1 andYY1-binding protein(Rybp). qRT-PCR verified that the mRNA levels of 4 genes increased during skeletal muscle differentiation. Rybp also increased in protein levels. Conclusions miRNA may promote muscle aging through interventing the processes such as transcription, metabolism and skeletal muscle differentiation. Rybp and its presumed regulatory miRNA miR-136 may promote the aging process of muscles.

Key words: muscle aging, miRNA, genes, skeletal muscle

中图分类号:

窦媛媛, 侯静雯, 王枚. 衰老过程中骨骼肌microRNA-mRNA调控网络分析[J]. 实用老年医学, 2022, 36(4): 354-359.

DOU Yuan-yuan, HOU Jing-wen, WANG Mei. Analysis of skeletal muscle microRNA-mRNA regulatory network during aging[J]. Practical Geriatrics, 2022, 36(4): 354-359.

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