miR‑148a inhibits pro‑inflammatory cytokines released by intervertebral disc cells by regulating the p38/MAPK pathway

Li,Guangfeng; Tang,Xianye; Chen,Hongliang; Sun,Wei; Yuan,Feng

doi:10.3892/etm.2018.6516

miR‑148a inhibits pro‑inflammatory cytokines released by intervertebral disc cells by regulating the p38/MAPK pathway

Authors:
- Guangfeng Li
- Xianye Tang
- Hongliang Chen
- Wei Sun
- Feng Yuan
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Affiliations: Graduate School of Xuzhou Medical College, Affiliated Hospital of Xuzhou Medical College, Xuzhou, Jiangsu 221000, P.R. China, Department of Spinal Surgery, Affiliated Hospital of Xuzhou Medical College, Xuzhou, Jiangsu 221000, P.R. China
Published online on: July 24, 2018 https://doi.org/10.3892/etm.2018.6516
Pages: 2665-2669

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Abstract

The present study aimed to verify the expression and investigate the role of microRNA (miR)‑148a in intervertebral disc degeneration (IDD) and explore the associated underlying mechanisms. Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) was performed to investigate levels of miR‑148a in the peripheral blood mononuclear cells (PBMCs) of patients with IDD. To investigate the role of miR‑148a in IDD, a stable miR‑148a‑overexpression/underexpression human nucleus pulposus (NP) cell line was generated by transfection with miR‑148a mimic/inhibitor. Then, NP cells were treated with LPS (10 µM) to induce inflammation. The mRNA expression level of miR‑148a in NP cells was determined by RT‑qPCR and the expression levels of p38 and p‑p38 were measured using western blotting. The mRNA expression and supernatant level of pro‑inflammatory cytokines, tumor necrosis factor (TNF‑α), interleukin (IL)‑1β and IL‑6, was evaluated by RT‑qPCR and ELISA, respectively. The results indicated that miR‑148a was significantly downregulated in the PBMCs of IDD patients compared with healthy controls. In vitro upregulation of miR‑148a in LPS‑stimulated NP cells, by transfection with miR‑148a mimic, resulted in inhibition of p‑p38 expression; however, inhibition of miR‑148a led to overexpression of p‑p38. Meanwhile, the production of pro‑inflammatory cytokines (TNF‑α, IL‑1β and IL‑6) was significantly reduced in miR‑148a‑overexpressing LPS‑stimulated NP cells and significantly increased in miR‑148a‑underexpressing NP cells. In conclusion, miR‑148a inhibits pro‑inflammatory cytokines released by intervertebral disc cells via regulation of the p38/mitogen‑activated protein kinase pathway.

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