Department of Sports Medicine, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, 266000, China.
Department of Abdominal ultrasound, The Affiliated Hospital of Qingdao University, Qingdao, Shandong, 266000, China.
BMC Musculoskelet Disord. 2024 Jun 8;25(1):457. doi: 10.1186/s12891-024-07568-x.
Type 2 diabetes mellitus (T2DM) is one of the high risk factors for sarcopenia. However, the pathogenesis of diabetic sarcopenia has not been fully elucidated. This study obtained transcriptome profiles of gastrocnemius muscle in normal and T2DM rats based on high-throughput sequencing technology, which may provide new ideas for exploring the pathogenesis of diabetic sarcopenia.
Twelve adult male Sprague-Dawley rats were randomly divided into Control group and T2DM group, and gastrocnemius muscle tissue was retained for transcriptome sequencing and real-time quantitative polymerase chain reaction (qRT-PCR) 6 months later. Screening differentially expressed genes (DEGs), Cluster analysis, gene ontology (GO) functional annotation analysis and Kyoto Encyclopedia of Genes and Gnomes (KEGG) functional annotation and enrichment analysis were performed for DEGs. Six DEGs related to apoptosis were selected for qTR-PCR verification.
Transcriptomic analysis showed that there were 1016 DEGs between the gastrocnemius muscle of T2DM and normal rats, among which 665 DEGs were up-regulated and 351 DEGs were down-regulated. GO analysis showed that the extracellular matrix organization was the most enriched in biological processes, with 26 DEGs. The extracellular matrix with 35 DEGs was the most abundant cellular component. The extracellular matrix structural constituent, with 26 DEGs, was the most enriched in molecular functions. The highest number of DEGs enriched in biological processes, cellular components and molecular functions were positive regulation of transcription by RNA polymerase II, nucleus and metal ion binding, respectively. There were 78, 230 and 89 DEGs respectively. KEGG pathway enrichment analysis showed that ECM-receptor interaction, PI3K-Akt signaling pathway and TGF-β signaling pathway(p < 0.001) had higher enrichment degree and number of DEGs. qRT-PCR results showed that the fold change of Map3k14, Atf4, Pik3r1, Il3ra, Gadd45b and Bid were 1.95, 3.25, 2.97, 2.38, 0.43 and 3.6, respectively. The fold change of transcriptome sequencing were 3.45, 2.21, 2.59, 5.39, 0.49 and 2.78, respectively. The transcriptional trends obtained by qRT-PCR were consistent with those obtained by transcriptome sequencing.
Transcriptomic analysis was used to obtain the "gene profiles" of gastrocnemius muscle of T2DM and normal rats. qRT-PCR verification showed that the genes related to apoptosis were differentially expressed. These DEGs and enrichment pathways may provide new ideas for exploring the pathogenesis of diabetic sarcopenia.
2 型糖尿病（T2DM）是肌少症的高危因素之一。然而，糖尿病性肌少症的发病机制尚未完全阐明。本研究基于高通量测序技术获得了正常和 T2DM 大鼠腓肠肌的转录组谱，这可能为探索糖尿病性肌少症的发病机制提供新的思路。
将 12 只成年雄性 Sprague-Dawley 大鼠随机分为对照组和 T2DM 组，6 个月后保留腓肠肌组织进行转录组测序和实时定量聚合酶链反应（qRT-PCR）。筛选差异表达基因（DEGs），进行聚类分析、基因本体（GO）功能注释分析和京都基因与基因组百科全书（KEGG）功能注释和富集分析。选择 6 个与细胞凋亡相关的 DEGs 进行 qTR-PCR 验证。
转录组分析显示，T2DM 大鼠与正常大鼠腓肠肌之间有 1016 个 DEGs，其中 665 个上调，351 个下调。GO 分析显示，生物过程中最丰富的是细胞外基质组织，有 26 个 DEGs。细胞外基质含有 35 个 DEGs，是最丰富的细胞成分。分子功能中最丰富的是细胞外基质结构成分，有 26 个 DEGs。生物过程、细胞成分和分子功能中富集程度最高的 DEGs 分别为 RNA 聚合酶 II 介导的转录正调控、核和金属离子结合。分别有 78、230 和 89 个 DEGs。KEGG 通路富集分析显示，细胞外基质受体相互作用、PI3K-Akt 信号通路和 TGF-β 信号通路（p<0.001）具有较高的富集程度和 DEGs 数量。qRT-PCR 结果显示，Map3k14、Atf4、Pik3r1、Il3ra、Gadd45b 和 Bid 的倍数变化分别为 1.95、3.25、2.97、2.38、0.43 和 3.6，转录组测序的倍数变化分别为 3.45、2.21、2.59、5.39、0.49 和 2.78。qRT-PCR 获得的转录趋势与转录组测序结果一致。
采用转录组分析获得 T2DM 和正常大鼠腓肠肌的“基因谱”。qRT-PCR 验证显示，与细胞凋亡相关的基因表达存在差异。这些 DEGs 和富集通路可能为探索糖尿病性肌少症的发病机制提供新的思路。
