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23 January 2017 : Laboratory Research  

Down-Regulation of miRNA-128 Contributes to Neuropathic Pain Following Spinal Cord Injury via Activation of P38

Zhaoyun Yang1BCDEF, Junmei Xu1BCDF, Rong Zhu1BCF, Lei Liu1ADE*

DOI: 10.12659/MSM.898788

Med Sci Monit 2017; 23:405-411

Abstract

BACKGROUND: Neuropathic pain (NPP) arises from a lesion or dysfunction of the somatosensory nervous system. Recent studies have demonstrated multiple microRNAs (miRNAs) play key roles in NPP development. This study aimed to investigate the effects of miR-128 on microglial cells.

MATERIAL AND METHODS: We established a compressive spinal cord injury (SCI) model and collected the spinal cord segment-derived conditioned medium (CM). We then measured the expression of miR-128 in the murine microglial cell line BV2 treated with CM-SCI or CM obtained from control (CM-NC). Furthermore, lentivirus production of miR-128 and scrambled control were transfected into BV2 cells, which were first treated with CM-SCI or CM-NC. Moreover, the effects of miR-128 on cell viability, M1/M2 microglial gene expression, inflammatory cytokines concentration, and the protein expression of P38 and phosphorylated P38 (P-P38) were investigated.

RESULTS: The expression of miR-128 was downregulated in murine microglial BV2 cells treated with CM-SCI. Overexpression of miR-128 markedly promoted the viability of murine microglial cells. In addition, miR-128 overexpression significantly decreased the expression levels of microglial M1 phenotypic markers CD86 and CD32, and increased the expression levels of M2 phenotypic markers Arg1 and CD206. Furthermore, miR-128 overexpression obviously decreased the concentration of TNF-α, IL-1β, and IL-6. We found that miR-128 overexpression significantly downregulated the expression levels of P38 andP-P38.

CONCLUSIONS: Our findings indicate that down-regulation of miR-128 in murine microglial cells may contribute to the development of NPP following SCI via activation of P38. MiR-128 may be a potential intervention target for NPP.

Keywords: Cell Survival - drug effects, Blotting, Western, Culture Media, Conditioned - pharmacology, Cytokines - metabolism, Down-Regulation - drug effects, Enzyme Activation - drug effects, Enzyme-Linked Immunosorbent Assay, Microglia - pathology, Neuralgia - pathology, Phosphorylation - drug effects, Real-Time Polymerase Chain Reaction, Spinal Cord Injuries - pathology, p38 Mitogen-Activated Protein Kinases - metabolism

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Medical Science Monitor Basic Research eISSN: 2325-4416
Medical Science Monitor Basic Research eISSN: 2325-4416