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Published on: August 10, 2018
MicroRNA‑138‑5p regulates the development of spinal cord injury by targeting SIRT1
1Department of Spine Surgery, The First People's Hospital of Lianyungang, Lianyungang, Jiangsu 222000, P.R. China.
Abstract:
MicroRNAs (miRs) play an important role in the development and progression of spinal cord injury (SCI). The role of miR‑138‑5p in SCI was investigated in the present study. The anti‑inflammatory effects of miR‑138‑5p and underlying mechanisms were investigated in an SCI rat model and in vitro model. Reverse transcription‑quantitative PCR (RT‑qPCR) was used to examine the expression of miR‑138‑5p in the SCI in vivo and in vitro models, as well as patients with SCI; it was found that miR‑138‑5p was significantly upregulated in SCI. Bioinformatics and dual‑luciferase reporter assays were performed to predict and confirm the binding sites between miR‑138‑5p and the 3'untranslated region of sirtuin 1 (SIRT1). Then, the expression of SIRT1 was detected via RT‑qPCR and western blotting, indicating downregulation of SIRT1 in SCI. PC12 cells were transfected with miR‑138‑5p inhibitor, inhibitor control or miR‑138‑5p inhibitor + SIRT1 small interfering RNA for 48 h, and then subjected to lipopolysaccharide (100 ng/ml) treatment for 4 h. Then, MTT assay, flow cytometry and ELISA experiments were performed to analyze cell viability, apoptosis, and the levels of tumor necrosis factor‑α, interleukin (IL)‑1β and IL‑6. Findings suggested that downregulation of miR‑138‑5p increased PC12 cell viability, inhibited cell apoptosis and attenuated proinflammatory responses, which may result in amelioration of SCI. However, all these effects were reversed by SIRT1 knockdown. Finally, it was observed that miR‑138‑5p altered the related protein expression of the PTEN/AKT pathway. These results indicated that miR‑138‑5p could regulate inflammatory responses and cell apoptosis in SCI models by modulating the PTEN/AKT signaling pathway via SIRT1, thus playing an important role in the development of SCI. Collectively, the present study demonstrated that miR‑138‑5p may be a novel therapeutic target for the treatment of SCI.
Insights
MicroRNA-138-5p is upregulated in spinal cord injury (SCI), promoting inflammation and cell death. Inhibiting this microRNA (miRNA) and targeting SIRT1 may offer new therapeutic strategies for SCI treatment.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- MicroRNAs (miRs) are critical regulators in the pathogenesis of spinal cord injury (SCI).
- The specific role and regulatory mechanisms of miR-138-5p in SCI remain incompletely understood.
Purpose of the Study:
- To investigate the role of miR-138-5p in the development and progression of spinal cord injury (SCI).
- To elucidate the anti-inflammatory effects and underlying molecular mechanisms of miR-138-5p in SCI.
Main Methods:
- Expression analysis of miR-138-5p and sirtuin 1 (SIRT1) using RT-qPCR and Western blotting in SCI models (in vivo, in vitro, and human patients).
- Bioinformatics and dual-luciferase reporter assays to confirm the interaction between miR-138-5p and SIRT1.
- Cellular assays (MTT, flow cytometry, ELISA) to assess cell viability, apoptosis, and inflammatory cytokine levels (TNF-α, IL-1β, IL-6) following transfection and treatment.
Main Results:
- miR-138-5p was significantly upregulated, while SIRT1 was downregulated in SCI models.
- Downregulation of miR-138-5p enhanced cell viability, reduced apoptosis, and attenuated pro-inflammatory responses in PC12 cells.
- These protective effects were reversed by SIRT1 knockdown, indicating SIRT1 mediates miR-138-5p's function via the PTEN/AKT pathway.
Conclusions:
- miR-138-5p exacerbates SCI by promoting inflammation and apoptosis, partly through modulating the PTEN/AKT pathway via SIRT1.
- Targeting miR-138-5p and its downstream effectors like SIRT1 presents a potential therapeutic avenue for managing spinal cord injury.
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