Research Paper
Upregulation of miR-125b-5p relieves chondrocyte inflammation and apoptosis in osteoarthritis by repressing the YAP1/NF-κB pathway
- 1 Department of Emergency Surgery, The First People's Hospital of Shangqiu City, Shangqiu 476000, Henan Province, China
- 2 Department of Orthopedics, Taizhou Hospital of Zhejiang Province, Taizhou 317000, Zhejiang Province, China
- 3 Operating Room, Taizhou Hospital of Zhejiang Province, Taizhou 317000, Zhejiang Province, China
Received: December 10, 2020 Accepted: May 31, 2021 Published: July 14, 2021
https://doi.org/10.18632/aging.203278How to Cite
Copyright: © 2021 Li et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Purpose: Presently, we aim to probe the influence of miR-125b-5p on osteoarthritis (OA) progression.
Methods: Interleukin-1β was applied for treating human chondrocytes to induce the OA model in vitro. The in-vivo OA model was constructed in rats by the destabilization of the medial meniscus (DMM). The profiles of miR-125b-5p, COX-2, iNOS, TNF-α, and IL-6 were compared by quantitative real-time polymerase chain reaction. The expression of Bax, Bcl2, Caspase3, COX-2, iNOS, Collagen II, Aggrecan, ADAM metallopeptidase with thrombospondin type 1 motif 5 (ADAMTS5), MMP9, YAP1 and NF-κB was measured by Western blotting. Cell viability and apoptosis were tested by the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay and flow cytometry. Safranin O, toluidine blue staining, immunohistochemistry (IHC) and OARSI standards were implemented to evaluate the OA severity and articular cartilage degradation. Griess reaction method determined the NO content, while TNF-α, IL-6, and PGE2 were quantified by enzyme-linked immunosorbent assay (ELISA). Bioinformatics analysis predicted the downstream targets of miR-125b-5p. The targeting relationship between YAP1 and NF-κB was verified by the dual-luciferase reporter assay and RNA pull-down assay.
Results: miR-125b-5p was downregulated in IL-1β-mediated OA. Overexpressing miR-125b-5p alleviated IL-1β-induced chondrocyte injury, hampered the profiles of COX-2, iNOS, TNF-α, IL-6, ADAMTS5, and MMP9, and strengthened the expression of Collagen II and Aggrecan. Mechanistically, miR-125b-5p targeted YAP1, and overexpressing miR-125b-5p repressed the YAP1/NF-κB pathway. In the in-vivo OA rat model, overexpressing miR-125b-5p eased inflammation and apoptosis in OA rats.
Conclusion: Overexpressing miR-125b-5p impeded inflammation and apoptosis in chondrocytes by repressing the YAP1/NF-κB pathway.
Abbreviations
ASG-IV: astragaloside IV; BMM: bone marrow macrophages; DMSO: discarded and methyl sulfoxide; EDTA: ethylenediamine tetraacetic acid; ECM: equilibrium of the extracellular matrix; ELISA: enzyme-linked immunosorbent assay; MSCs: mesenchymal stem cells; OA: Osteoarthritis; qRT-PCR: quantitative real-time polymerase chain reaction.