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MiR-19b-3p attenuates IL-1β induced extracellular matrix degradation and inflammatory injury in chondrocytes by targeting GRK6

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Abstract

Osteoarthritis (OA) is characterized by degradation of articular cartilage. MiRNAs are involved in the regulation of chondrogenesis and OA. We aimed to investigate effects and mechanisms of miR-19b-3p in regulating chondrocytes viability, cartilage degradation and inflammatory response. Primary chondrocytes were isolated from cartilages in control subjects and patients with OA. Murine ATDC5 cells were pre-conditioned with IL-1β in vitro. Expressions and interaction of miR-19b-3p with G protein-coupled receptor kinase 6 (GRK6), and their effects on inflammation, chondrocytes viability and cartilage degradation were determined after miR-19b-3p mimic or GRK6 siRNA transfection. MiR-19b-3p was significantly decreased in OA chondrocytes and IL-1β-stimulated ATDC5 cells, in paralleled with the elevated type-II-collagen, aggrecan, MMP13 and GRK6 expression. MiR-19b-3p mimic dramatically increased the viability of chondrocytes and suppressed cell apoptosis. It also increased type-II-collagen, aggrecan expression and glycosaminoglycan (sGAG) content, and decreased the expression of MMP-1 and MMP-13 that controlled by IL-1β. Overexpression of miR-19b-3p inhibited the production of IL-6 and IL-8 in ATDC5 cells. However, the protective effects of miR-19b-3p mimic on IL-1β induced cell death; IL-8 production and sGAG decrease were greatly discounted by GRK6 lentiviral vectors. Luciferase reporter assay confirmed that GRK6 gene was a direct target ofmiR-19b-3p. GRK6 siRNA transfection antagonized the IL-1β-induced chondrocytes injury, extracellular matrix degradation and inflammatory response. MiR-19b-3p mimic and GRK6 siRNA showed comparable inhibitory effect on IL-1β-provoked NF-κB as reflected by the expression of p-p65. NF-κB translocation inhibition with PS1154 reversed the effects of IL-1β on IL-8 and sGAG. Collectively, miR-19b-3p attenuated OA by targeting GRK6-NF-κB pathway.

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Funding

This work is supported by Shaanxi Science and Technology Co-ordination & Innovation Project (Grant No. 2015KTCL03-02).

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Authors and Affiliations

  1. Department of Orthopaedics, Shaanxi Provincial People’s Hospital, NO. 256 Youyi Xi Road, Xi’an, 710068, Shaanxi, China

    Liang Duan, Dapeng Duan, Wenbo Wei, Zhengming Sun, Honghai Xu, Lipeng Guo & Xuanhui Wu

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  1. Liang Duan
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  2. Dapeng Duan
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  3. Wenbo Wei
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  4. Zhengming Sun
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  5. Honghai Xu
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Correspondence to Honghai Xu.

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This study was approved by the ethics committee of the Shaanxi Provincial People’s Hospital and adhered to the Helsinki Declaration (2000).

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Duan, L., Duan, D., Wei, W. et al. MiR-19b-3p attenuates IL-1β induced extracellular matrix degradation and inflammatory injury in chondrocytes by targeting GRK6. Mol Cell Biochem 459, 205–214 (2019). https://doi.org/10.1007/s11010-019-03563-2

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  • DOI: https://doi.org/10.1007/s11010-019-03563-2

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