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  • Title: MiR-146a-5p promotes IL-1β-induced chondrocyte apoptosis through the TRAF6-mediated NF-kB pathway.
    Author: Shao J, Ding Z, Peng J, Zhou R, Li L, Qian Q, Chen Y.
    Journal: Inflamm Res; 2020 Jun; 69(6):619-630. PubMed ID: 32328683.
    Abstract:
    OBJECTIVE: This study aimed to explore the role of the miR-146a-5p/TRAF6/NF-KB axis in chondrocyte apoptosis. METHODS: Transcriptome sequencing for microRNA expression in control and osteoarthritic cartilage was performed. Bioinformatic analysis was performed to identify the target genes of miR-146a-5p, and subsequently, Gene Ontology (GO) terms and KEGG pathways were identified. Furthermore, protein-protein interactions were analyzed to identify the hub regulatory gene of miR-146a-5p. MiR-146a-5p mimic, inhibitor and the corresponding negative control were constructed, and the apoptosis rates were measured in the transfected groups by flow cytometry, TUNEL staining and Western blot. Potential miRNA-target interactions were identified by dual-luciferase reporter assay. RESULTS: The microRNA array demonstrated that miR-146a-5p was significantly upregulated in osteoarthritic tissues, which was further confirmed by PCR analysis. Compared with the control group, IL-1β significantly decreased the viability of chondrocytes, while coculture with miR-146a-5p inhibitor rescued the IL-1β-induced inhibition of chondrocyte viability. Western blot results also identified the proapoptotic effects of miR-146a-5p. Bioinformatic analysis results revealed that miR-146a-5p targeted 159 potential genes, and TRAF6 was the hub gene among the 159 genes. The relative expression of TRAF6 was significantly decreased in the IL-1β-induced group. When siTRAF6 was added, apoptosis was significantly increased. Luciferase reporter assays showed that luciferase activity of the TRAF6 3'-UTR reporter was decreased in chondrocytes after transfection with the miR-146a-5p mimic. CONCLUSIONS: This work showed that miR-146 induces chondrocyte apoptosis by targeting the TRAF6-mediated NF-KB signaling pathway, and miR-146 may be a potential target for OA treatment.
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