基于高通量测序技术构建糖尿病肾病小鼠lncRNA相关ceRNA调控网络

Construction of lncRNA-related ceRNA regulatory network of diabetic kidney disease mouse based on high-throughput sequencing technology

目的:筛选糖尿病肾病(DKD)小鼠肾脏组织中差异表达的长链非编码RNA(lncRNA)、小RNA(miRNA),构建lncRNA相关竞争性内源RNA(ceRNA)调控网络。方法:选取6只BKS-db/m雄性小鼠为对照组,6只BKS-db/db雄性小鼠为模型组,模型组构建DKD小鼠模型。处死小鼠后取肾脏组织进行高通量测序,利用DESeq软件筛选两组差异表达的lncRNA、miRNA,使用Miranda软件进行差异表达的miRNA靶基因预测。构建lncRNA相关ceRNA调控网络并进行GO功能富集分析以及KEGG信号通路富集分析。结果:共筛选出DKD小鼠差异表达的lncRNA 1495个、差异表达的miRNA 72个。成功构建由MSTRG.7252.3、MSTRG.10465.2、MSTRG.16253.3等23个lncRNA、23个miRNA与2个mRNA组成,与lncRNA相关的ceRNA网络。GO功能富集分析显示,该网络主要涉及生物学过程、细胞组成和分子功能;KEGG信号通路富集显示,主要与PPAR信号通路、造血细胞谱系、细胞黏附分子、TNF信号通路等有关。结论:成功构建DKD小鼠lncRNA相关的ceRNA调控网络。

Aim:To screen the differentially expressed long non-coding RNA(lncRNA)and microRNA(miRNA)in the kidney tissue of diabetic kidney disease(DKD)mice and construct a lncRNA-related competing endogenous RNA(ceRNA)regulatory network.Methods:A total of 6 BKS-db/m male mice were selected as control group,and 6 BKS-db/db male mice were selected as model group to establish the DKD mouse model.The mice were sacrificed and their kidney tissue samples were collected for high-throughput sequencing.The differentially expressed lncRNAs and miRNAs between the 2 groups were screened using DESeq software.The target genes of differentially expressed miRNAs were predicted using Miranda software.A lncRNA-related ceRNA regulatory network was constructed,and GO functional enrichment analysis and KEGG signaling pathway enrichment analysis were performed.Results:A total of 1495 differentially expressed lncRNAs and 72 differentially expressed miRNAs were screened.A lncRNA-related ceRNA network consisting of 23 lncRNAs such as MSTRG.7252.3,MSTRG.10465.2,MSTRG.16253.3,etc,23 miRNAs,and 2 mRNAs was successfully constructed.The GO functional enrichment analysis showed that the main categories involved biological processes,cellular components,and molecular functions.The KEGG signaling pathway enrichment analysis revealed that the pathways were primarily related to PPAR signaling pathway,hematopoietic cell lineage,cell adhesion molecules,TNF signaling pathway,etc.Conclusion:A lncRNA-related ceRNA regulatory network in DKD mice has been successfully constructed.