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Updated: Sep 18, 2025
Quantitative Real-Time PCR Evaluation of microRNA Expressions in Mouse Kidney with Unilateral Ureteral Obstruction
Published on: August 27, 2020
Identification of core noncoding RNA-associated competing endogenous RNA networks in renal fibrosis via
Yizhen Chen1, Meng Cheng2, Rong Dai2
1First Clinical Medical College, Anhui University of Chinese Medicine, Hefei, China.
Objective:
This study aimed to investigate noncoding RNA (ncRNA) expression changes in renal fibrosis (RF) models induced by three distinct etiologies using whole-transcriptome RNA sequencing, identify overlapping differentially expressed (DE) ncRNAs, construct core competing endogenous RNA (ceRNA) networks, and explore their role in RF.
Methods:
Three RF rat models, 5/6 nephrectomy, adenine, and unilateral ureteral obstruction, were established. DE RNAs were identified through sequencing and validated by real-time quantitative polymerase chain reaction. ceRNA and RNA-binding protein (RBP) networks were visualized using Cytoscape. Core ceRNA axes were validated with dual-luciferase assay, RNA fluorescence in situ hybridization, western blot, immunofluorescence, and immunohistochemistry. Enrichment analysis was performed to explore potential functions.
Results:
Sequencing analysis revealed significant dysregulation of ncRNAs in all models compared to the normal group. Intersection analysis identified 215 mRNAs, 19 lncRNAs, and 247 circRNAs as overlapping DE RNAs. lncRNA-based ceRNA networks comprising 7 lncRNAs, 8 miRNAs, and 21 mRNAs, and circRNA-based networks comprising 13 circRNAs, 29 miRNAs, and 41 mRNAs were constructed. The TCONS_00008870/circRNA_3140-miR-466b-3p-Adamts2 axis was identified as a key regulatory pathway. Enrichment analysis showed significant pathways including Rap1 signaling, extracellular matrix-receptor interaction, and PI3K-Akt signaling, with RBPs enriched in RNA binding and ferroptosis.
Conclusion:
By integrating data from three distinct models, we identified conserved ceRNA axis-TCONS_00008870/circRNA_3140-miR-466b-3p-Adamts2-potentially modulating fibrotic progression in renal tissue.
