长链非编码RNA-微小RNA-信使RNA网络分析揭示隔药灸治疗克罗恩病大鼠的潜在生物标志物
LncRNA-miRNA-mRNA Network Analysis Reveals the Potential Biomarkers in Crohn's Disease Rats Treated with Herb-Partitioned Moxibustion.
作者信息
Wang Xue-Jun, Li Xiao-Ying, Guo Xiao-Cong, Liu Li, Jin You-You, Lu Yun-Qiong, Cao Yao-Jia-Ni, Long Jun-Yi, Wu Huan-Gan, Zhang Dan, Yang Guang, Hong Jue, Yang Yan-Ting, Ma Xiao-Peng
机构信息
Yueyang Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai University of Traditional Chinese Medicine, Shanghai, People's Republic of China.
Key Laboratory of Acupuncture-Moxibustion and Immunology, Shanghai Research Institute of Acupuncture and Meridian, Shanghai, People's Republic of China.
出版信息
J Inflamm Res. 2022 Mar 5;15:1699-1716. doi: 10.2147/JIR.S351672. eCollection 2022.
BACKGROUND
Long noncoding RNA (lncRNA) is receiving growing attention in Crohn's disease (CD). However, the mechanism by which herb-partitioned moxibustion (HPM) regulates the expression and functions of lncRNAs in CD rats is still unclear. The aim of our study is to identify lncRNA-miRNA-mRNA network potential biological functions in CD.
METHODS
RNA sequencing and microRNA (miRNA) sequencing were carried out to analyze lncRNA, miRNA and mRNA expression profiles among the CD rats, normal control rats, and CD rats after HPM treatment and constructed the potential related lncRNA-miRNA-mRNA competing endogenous RNA (ceRNA) networks. Then, Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, protein-protein interaction (PPI) analysis and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to explore potentially important genes in ceRNA networks.
RESULTS
A total of 189 lncRNAs, 32 miRNAs and 463 mRNAs were determined as differentially expressed (DE) genes in CD rats compared to normal control rats, and 161 lncRNAs, 12 miRNAs and 130 mRNAs were identified as remarkably DE genes in CD rats after HPM treatment compared to CD rats. GO analysis indicated that the target genes were most enriched in cAMP and in KEGG pathway analysis the main pathways included adipocytokine, PPAR, AMPK, FoxO and PI3K-Akt signaling pathway. Finally, qRT-PCR results confirmed that lncRNA LOC102550026 sponged miRNA-34c-5p to regulate the intestinal immune inflammatory response by targeting Pck1.
CONCLUSION
By constructing a ceRNA network with lncRNA-miRNA-mRNA, PCR verification, and KEGG analysis, we revealed that LOC102550026/miRNA-34c-5p/Pck1 axis and adipocytokine, PPAR, AMPK, FoxO, and PI3K-Akt signaling pathways might regulate the intestinal immune-inflammatory response, and HPM may regulate the lncRNA LOC102550026/miR-34c-5p/Pck1 axis and adipocytokine, PPAR, AMPK, FoxO, and PI3K-Akt signaling pathways, thus improving intestinal inflammation in CD. These findings may be novel potential targets in CD.
背景
长链非编码RNA（lncRNA）在克罗恩病（CD）中受到越来越多的关注。然而，隔物灸（HPM）调节CD大鼠lncRNAs表达和功能的机制仍不清楚。我们研究的目的是确定CD中lncRNA-miRNA-mRNA网络的潜在生物学功能。
方法
进行RNA测序和微小RNA（miRNA）测序，以分析CD大鼠、正常对照大鼠以及HPM治疗后的CD大鼠之间的lncRNA、miRNA和mRNA表达谱，并构建潜在相关的lncRNA-miRNA-mRNA竞争性内源RNA（ceRNA）网络。然后，进行基因本体论（GO）、京都基因与基因组百科全书（KEGG）通路富集分析、蛋白质-蛋白质相互作用（PPI）分析和定量实时聚合酶链反应（qRT-PCR），以探索ceRNA网络中潜在的重要基因。
结果
与正常对照大鼠相比，共确定189个lncRNAs、32个miRNAs和463个mRNAs为CD大鼠中的差异表达（DE）基因；与CD大鼠相比，161个lncRNAs、12个miRNAs和130个mRNAs被鉴定为HPM治疗后CD大鼠中的显著DE基因。GO分析表明，靶基因在cAMP中富集最多，KEGG通路分析中主要通路包括脂肪细胞因子、PPAR、AMPK、FoxO和PI3K-Akt信号通路。最后，qRT-PCR结果证实lncRNA LOC102550026通过靶向Pck1来海绵化miRNA-34c-5p以调节肠道免疫炎症反应。
结论
通过构建lncRNA-miRNA-mRNA的ceRNA网络、PCR验证和KEGG分析，我们揭示了LOC102550026/miRNA-34c-5p/Pck1轴以及脂肪细胞因子、PPAR、AMPK、FoxO和PI3K-Akt信号通路可能调节肠道免疫炎症反应，且HPM可能调节lncRNA LOC102550026/miR-34c-5p/Pck1轴以及脂肪细胞因子、PPAR、AMPK、FoxO和PI3K-Akt信号通路，从而改善CD中的肠道炎症。这些发现可能是CD中的新型潜在靶点。
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