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目的筛选慢性宫内缺氧(CIH)子代大鼠骨骼肌差异表达基因(DEGs)并进行生物信息学分析。方法 20只孕鼠随机分为宫内缺氧组及常氧组,取新生雄鼠股四头肌行数字化基因表达谱(DGE)测序筛选DEGs,进行Gene Ontology及KEGG Pathway分析。实时荧光定量PCR验证测序结果可靠性。结果 1测序reads分布、饱和度及随机性显示测序质量良好。2CIH后DEGs达396个,其中上调表达基因287个,下调109个。GO功能注释表明,DEGs主要集中在细胞组分、胞外区、封闭的膜腔等部位,具有结合、催化、转运等活性,参与发育过程、代谢过程、生物调节等多个生物学过程。Pathway富集分析显示DEGs主要集中在三大营养物质代谢、Insulin信号通路、PPAR信号转导通路。3实时荧光定量PCR验证DEGs上调或下调趋势与DGE结果基本一致。结论 CIH能程序性控制子鼠骨骼肌基因表达,其中糖脂代谢调节是CIH相关的成年期慢性代谢性疾病的主要分子基础。
Objective To screen differentially expressed genes (DEGs) of skeletal muscle of chronic intrauterine hypoxia (CIH) offspring rats and analyze their bioinformatics. Methods Twenty pregnant rats were randomly divided into intrauterine hypoxia group and normoxia group. The degenerative quadriceps of newborn male rats were screened by digital gene expression profile (DGE) for DEGs, and then analyzed by Gene Ontology and KEGG Pathway. Real-time fluorescence quantitative PCR to verify the reliability of sequencing results. Results 1 Sequencing reads distribution, saturation and randomness showed sequencing quality was good. There were 396 DEGs after 2CIH, including 287 up-regulated genes and 109 down-regulated genes. GO functional annotation indicates that DEGs are mainly concentrated in cellular components, extracellular regions and closed membranous cavities, etc. They have the activity of binding, catalysis and transport, and are involved in many biological processes such as developmental process, metabolic process and biological regulation. Pathway enrichment analysis showed that DEGs mainly concentrated on three major nutrient metabolism, Insulin signaling pathway, PPAR signaling pathway. 3 real-time quantitative PCR confirmed DEGs up or down trend and DGE results are basically the same. Conclusion CIH can control the gene expression of skeletal muscle of rats in a programmed manner. The regulation of glucose and lipid metabolism is the main molecular basis of CIH-related chronic metabolic diseases in adulthood.