Zhuang Xing-Xing, Liu Tao, Wei Liang-Bing, Gao Jia-Rong
Department of Pharmacy, Chaohu Hospital of Anhui Medical University, Chaohu, Anhui 238000, P.R. China.
Department of Pharmacy, The First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei, Anhui 230012, P.R. China.
Exp Ther Med. 2023 Jul 7;26(2):403. doi: 10.3892/etm.2023.12102. eCollection 2023 Aug.
Long non-coding RNAs (lncRNAs) are ncRNA transcripts >200 nucleotides that are important genetic regulators. LncRNAs can directly regulate mRNA through a lncRNA-mRNA regulatory mode and can also regulate mRNA through competitive binding to micro (mi)RNA, which is generally known as the competitive endogenous RNA (ceRNA) network. The present study evaluated the functional roles and regulatory networks of lncRNAs in chronic glomerulonephritis (CGN). The proliferative ability of mouse glomerular mesangial cells (GMCs) induced by different concentrations of lipopolysaccharide (LPS) was assessed using the Cell Counting Kit-8 assay, and RNA sequencing (RNA-seq) was performed to identify differentially expressed lncRNAs in LPS-induced GMCs. Based on the sequencing results, six lncRNAs were selected for validation using reverse transcription-quantitative PCR (RT-qPCR). Furthermore, the lncRNA-mRNA regulatory network and the lncRNA-miRNA-mRNA ceRNA network were constructed to assess the role and mechanism of CGN-related lncRNAs. To elucidate the biological functions of lncRNAs, Gene Ontology (GO) biological process term enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were performed on all mRNAs involved in the lncRNA-mRNA regulatory network and in the ceRNA network. A total of 1,532 differentially expressed lncRNAs, including 594 upregulated lncRNAs and 938 downregulated lncRNAs, were identified using RNA-seq. The results of RT-qPCR validation were consistent with RNA-seq results. An lncRNA-mRNA regulatory network, including 236 lncRNAs and 556 mRNAs, and a ceRNA network, including 6 lncRNAs, 18 miRNAs and 419 mRNAs, were successfully constructed. The GO biological process term enrichment and KEGG pathway enrichment analyses demonstrated that those lncRNAs were often related to inflammatory response and substance metabolism. The present study identified key CGN-related lncRNAs in LPS-induced GMCs, and further demonstrated a global view of the lncRNA-mRNA regulatory network and ceRNA network involved in CGN. These results offered novel insights into the roles of lncRNAs in the pathogenesis of CGN and identified potential diagnostic biomarkers.
长链非编码RNA(lncRNAs)是长度大于200个核苷酸的非编码RNA转录本,是重要的基因调控因子。LncRNAs可通过lncRNA-mRNA调控模式直接调控mRNA,也可通过与微小(mi)RNA竞争性结合来调控mRNA,这一过程通常被称为竞争性内源RNA(ceRNA)网络。本研究评估了lncRNAs在慢性肾小球肾炎(CGN)中的功能作用和调控网络。使用细胞计数试剂盒-8检测法评估不同浓度脂多糖(LPS)诱导的小鼠肾小球系膜细胞(GMCs)的增殖能力,并进行RNA测序(RNA-seq)以鉴定LPS诱导的GMCs中差异表达的lncRNAs。基于测序结果,选择6个lncRNAs使用逆转录定量PCR(RT-qPCR)进行验证。此外,构建lncRNA-mRNA调控网络和lncRNA-miRNA-mRNA ceRNA网络,以评估与CGN相关的lncRNAs的作用和机制。为阐明lncRNAs的生物学功能,对参与lncRNA-mRNA调控网络和ceRNA网络的所有mRNA进行基因本体论(GO)生物学过程术语富集分析和京都基因与基因组百科全书(KEGG)通路富集分析。使用RNA-seq共鉴定出1532个差异表达的lncRNAs,其中包括594个上调的lncRNAs和938个下调的lncRNAs。RT-qPCR验证结果与RNA-seq结果一致。成功构建了一个包含236个lncRNAs和556个mRNAs的lncRNA-mRNA调控网络,以及一个包含6个lncRNAs、18个miRNAs和419个mRNAs的ceRNA网络。GO生物学过程术语富集分析和KEGG通路富集分析表明,这些lncRNAs通常与炎症反应和物质代谢相关。本研究在LPS诱导的GMCs中鉴定出与CGN相关的关键lncRNAs,并进一步展示了参与CGN的lncRNA-mRNA调控网络和ceRNA网络的整体情况。这些结果为lncRNAs在CGN发病机制中的作用提供了新的见解,并鉴定出潜在的诊断生物标志物。