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miR-486-5p Inhibits Inflammatory Response, Matrix Degradation and Apoptosis of Nucleus Pulposus Cells through

Xingyu Chai1,2, Haipeng Si3, Jiang Song2

  • 1Department of Traumatology, Shandong Provincial Qianfoshan Hospital, Shandong University, Jinan, China.

Cellular Physiology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology
|February 22, 2019
PubMed
Abstract

Insights

microRNA-486-5p (miR-486-5p) inhibits intervertebral disc degeneration (IDD) by targeting FOXO1. This study reveals miR-486-5p protects nucleus pulposus cells from inflammation and apoptosis, offering potential IDD therapies.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Intervertebral disc degeneration (IDD) is a significant health issue.
  • microRNA-486-5p (miR-486-5p) and forkhead box protein O1 (FOXO1) are implicated in IDD pathogenesis.
  • The precise molecular mechanisms involving miR-486-5p and FOXO1 in IDD are not fully understood.

Purpose of the Study:

  • To elucidate the molecular mechanisms of miR-486-5p and FOXO1 in intervertebral disc degeneration (IDD).
  • To investigate the regulatory relationship between miR-486-5p and FOXO1 in nucleus pulposus (NP) cells.

Main Methods:

  • Quantitative real-time PCR (qRT-PCR) to assess gene expression.
  • In-vitro cell culture and transfection techniques.
  • Cell Counting Kit-8 (CCK-8) assay for cell viability.
  • Flow cytometry for apoptosis analysis.
  • Luciferase reporter assays to confirm direct targeting.
  • Western blotting to measure protein expression of inflammatory and matrix-degrading factors.

Main Results:

  • miR-486-5p expression was significantly downregulated, while FOXO1 was upregulated in lipopolysaccharide (LPS)-stimulated NP cells.
  • Overexpression of miR-486-5p suppressed inflammatory cytokines (IL-1β, IL-6, TNF-α) and matrix degrading enzymes (MMP-3, MMP-13, ADAMTS-4, ADAMTS-5).
  • miR-486-5p promoted the expression of extracellular matrix (ECM) genes (Aggrecan, Collagen II) and protected NP cells against LPS-induced apoptosis.
  • FOXO1 was identified as a direct target of miR-486-5p; its overexpression exacerbated LPS-induced injury and counteracted miR-486-5p's protective effects.

Conclusions:

  • miR-486-5p exerts protective effects against IDD by directly targeting FOXO1.
  • miR-486-5p inhibits inflammatory response, ECM degradation, and apoptosis in NP cells.
  • Targeting the miR-486-5p/FOXO1 axis presents a potential therapeutic strategy for IDD.
Keywords:
FOXO1IDDNPmiR-486-5p

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