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LncRNA H19 Regulates Lipopolysaccharide (LPS)-Induced Apoptosis and Inflammation of BV2 Microglia Cells Through Targeting miR-325-3p/NEUROD4 Axis

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Abstract

Spinal cord injury (SCI) is a devastating traumatic event worldwide. Work from the past decade has highlighted the key involvement of long non-coding RNAs (lncRNAs) in SCI. Nevertheless, the molecular action of lncRNA H19 in SCI is still not fully understood. The levels of H19, microRNA (miR)-325-3p, and neuronal differentiation 4 (NEUROD4) were determined by quantitative real-time polymerase chain reaction (qRT-PCR) or western blot. Flow cytometry was performed to assess cell apoptosis. The levels of tumor necrosis factor-α (TNF-α), interleukin 1β (IL-1β), and IL-6 were detected using the enzyme-linked immunosorbent assay (ELISA). Targeted relationships among H19, miR-325-3p, and NEUROD4 were confirmed by dual-luciferase reporter, RNA immunoprecipitation (RIP), or RNA pull-down assays. Our data showed that H19 level was overexpressed in lipopolysaccharide (LPS)-treated BV2 cells. H19 silencing alleviated LPS-evoked cell apoptosis and inflammation. Mechanistically, H19 in BV2 cells directly targeted miR-325-3p, and NEUROD4 was a direct target of miR-325-3p. Moreover, miR-325-3p was a functional target of H19 in regulating cell apoptosis and inflammation induced by LPS. Enforced expression of miR-325-3p relieved LPS-evoked cell apoptosis and inflammation through reducing NEUROD4. Furthermore, H19 in BV2 cells regulated NEUROD4 expression through targeting miR-325-3p. Our results identified that the silencing of H19 attenuated LPS-evoked microglia cell apoptosis and inflammation after SCI at least partially through targeting the miR-325-3p/NEUROD4 axis, highlighting a novel approach for SCI management.

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Data Availability

All data generated or analyzed during this study are included in this published article.

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Funding

The present study was supported by Fuzhou Science and Technology Project (2019-SZ-24).

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Authors and Affiliations

  1. Department of Orthopedics, Fuzhou Second Hospital Affiliated To Xiamen University, Cangshan District, 47 Shangteng Road, Fuzhou, 350007, Fujian, China

    Enyi Gu, Weikun Pan, Kangyao Chen, Zhong Zheng, Guoling Chen & Pengde Cai

Authors
  1. Enyi Gu
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  2. Weikun Pan
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  3. Kangyao Chen
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  4. Zhong Zheng
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  5. Guoling Chen
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  6. Pengde Cai
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Contributions

Enyi Gu and Weikun Pan conceived and designed the experiments; Kangyao Chen performed the experiments; Zhong Zheng contributed reagents/materials/analysis tools; Guoling Chen and Pengde Cai wrote the paper. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Pengde Cai.

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The authors declare that they have no financial conflicts of interest.

Ethics Approval and Consent to Participate

The study was approved by the Ethics Committee of Fuzhou Second Hospital Affiliated to Xiamen University, and all experimental procedures were done following the Chinese National Guide for the Care and Use of Laboratory Animals.

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Informed consent was obtained from all patients.

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Highlights: (1) miR-325-3p was a functional target of H19 in regulating cell apoptosis and inflammation induced by LPS; (2) H19 regulated NEUROD4 expression through targeting miR-325-3p; (3) H19 silencing attenuated LPS-evoked microglia cell apoptosis and inflammation by targeting the miR-325-3p/NEUROD4 axis.

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Gu, E., Pan, W., Chen, K. et al. LncRNA H19 Regulates Lipopolysaccharide (LPS)-Induced Apoptosis and Inflammation of BV2 Microglia Cells Through Targeting miR-325-3p/NEUROD4 Axis. J Mol Neurosci 71, 1256–1265 (2021). https://doi.org/10.1007/s12031-020-01751-0

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  • DOI: https://doi.org/10.1007/s12031-020-01751-0

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