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Published on: May 21, 2018
Long non-coding RNA Mirt2 relieves lipopolysaccharide-induced injury in PC12 cells by suppressing miR-429
Haibo Li1, Yu Xu1, Guoxiu Wang2
1Department of Orthopaedics, Shaoxing People's Hospital (Shaoxing Hospital of Zhejiang University), Shaoxing, 312000, China.
Abstract:
Long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) play important roles in the pathogenesis of spinal cord injury (SCI). This study investigated the effects of lncRNA Mirt2 and miR-429 on lipopolysaccharide (LPS)-induced injuries in PC12 cells. Serum samples were collected from 36 patients with SCI and the healthy controls. The expression of lncRNA Mirt2 in serum samples was measured by qRT-PCR. The in vitro model of SCI was established by treating PC12 cells with LPS. The effects of lncRNA Mirt2 and miR-429 on the cell model were evaluated by CCK-8 assay, flow cytometry, western blot, qRT-PCR, and ELISA. Further, the activation of NF-κB and p38MAPK pathways was tested by western blot. LPS induced obvious cell injuries in PC12 cells, as cell viability was reduced, apoptosis rate was increased, caspase-3 and -9 were cleaved, and the release of TNF-α and IL-6 was induced. lncRNA Mirt2 was up-regulated in LPS-stimulated PC12 cells and serum samples derived from SCI patients. Overexpression of lncRNA Mirt2 protected PC12 cells against LPS-induced injuries. Further studies found that lncRNA Mirt2 acted as the molecular sponge of miR-429 and miR-34a-5p. lncRNA Mirt2 did not protect PC12 cells when miR-429 was overexpressed. Moreover, the inhibitory effects of lncRNA Mirt2 on NF-κB and p38MAPK pathways were abolished when miR-429 was overexpressed. lncRNA Mirt2 exerts protective effects in an in vitro model of SCI by down-regulating miR-429. This study shed light on the treatment of SCI by using the lncRNA-miRNA regulation network.
Insights
Long non-coding RNA Mirt2 protects against spinal cord injury (SCI) by down-regulating miR-429, offering new therapeutic insights. This study highlights the lncRNA-miRNA network
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- Long non-coding RNAs (lncRNAs) and microRNAs (miRNAs) are implicated in spinal cord injury (SCI) pathogenesis.
- Understanding the regulatory roles of specific lncRNAs and miRNAs is crucial for developing effective SCI treatments.
Purpose of the Study:
- To investigate the role of lncRNA Mirt2 and miR-429 in lipopolysaccharide (LPS)-induced neuronal injury.
- To elucidate the underlying molecular mechanisms of lncRNA Mirt2 in an in vitro SCI model.
Main Methods:
- Established an in vitro SCI model using LPS-treated PC12 cells.
- Quantified lncRNA Mirt2 and miR-429 expression via qRT-PCR.
- Assessed cell viability (CCK-8), apoptosis (flow cytometry), and inflammatory markers (ELISA).
- Analyzed NF-κB and p38MAPK pathway activation using Western blot.
Main Results:
- LPS induced significant PC12 cell injury, increasing apoptosis and inflammatory cytokine release (TNF-α, IL-6).
- lncRNA Mirt2 was upregulated in LPS-treated cells and SCI patient serum.
- Overexpression of lncRNA Mirt2 protected cells against LPS-induced injury and inhibited NF-κB/p38MAPK pathways.
- lncRNA Mirt2 acted as a molecular sponge for miR-429, and its protective effects were dependent on miR-429 levels.
Conclusions:
- lncRNA Mirt2 exerts protective effects in an in vitro SCI model by down-regulating miR-429.
- The lncRNA-miRNA regulatory network, specifically lncRNA Mirt2/miR-429, presents a potential therapeutic target for SCI.
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