Protective effect and mechanism of microRNA‑146a on ankle fracture

Mao,Haijun; Xu,Guangyue

doi:10.3892/etm.2020.9131

Protective effect and mechanism of microRNA‑146a on ankle fracture

Authors:
- Haijun Mao
- Guangyue Xu
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Affiliations: Department of Orthopedics, The Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing, Jiangsu 210008, P.R. China
Published online on: August 25, 2020 https://doi.org/10.3892/etm.2020.9131
Article Number: 3
Copyright: © Mao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The present study investigated the expression and role of microRNA‑146a (miR‑146a) on ankle fracture, and explored the underlying mechanism. miR‑146a levels in the blood of patients with ankle fracture was measured using reverse transcription‑quantitative PCR (RT‑qPCR). Expression of pro‑inflammatory factors in the peripheral blood of ankle fracture patients was also detected using ELISA. Oxidative stress biomarkers including malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) were additionally investigated. The role of miR‑146a in ankle fracture was investigated in vitro where MG‑63 cells were transfected with miR‑146a mimic or miR‑146a inhibitor for 2 h, then treated with 1 µg/ml bradykinin for 24 h. An MTT assay was then performed to assess cell viability and pro‑inflammatory factors were detected via RT‑qPCR and western blot analysis. Finally, activation of the TNF receptor associated factor 6 (TRAF6)/NF‑κB pathway was determined via western blotting and RT‑qPCR. The results demonstrated that miR‑146a was significantly downregulated in the blood of patients with ankle fracture. The protein levels of tumor necrosis factor (TNF‑α), interleukin (IL)‑1β and IL‑6 were significantly upregulated in patients with ankle fracture. In addition, MDA content significantly increased, and SOD and CAT activity significantly decreased in patients with ankle fracture. In vitro experiments demonstrated that miR‑146a overexpression significantly enhanced cell viability. miR‑146a mimic suppressed BK‑induced upregulation of TNF‑α, IL‑1β, IL‑6 and MDA, and increased SOD and CAT activity. Finally, miR‑146a mimic inhibited activation of the TRAF6/NF‑κB pathway whilst miR‑146a inhibitor had the opposite effect. In conclusion, miR‑146a may be a potential therapeutic target for the treatment of ankle fracture by inhibiting the inflammatory response and attenuating oxidative stress.

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