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Protective Effect of miR-374a on Chemical Hypoxia-Induced Damage of PC12 Cells In Vitro via the GADD45α/JNK Signaling Pathway

Neurochemical Research volume 43, pages 581–590 (2018)Cite this article

Abstract

To explore the effect of microRNA-374a (miR-374a) on chemical hypoxia-induced pheochromocytoma (PC12) cell damage by mediating growth arrest and the DNA damage-45 alpha (GADD45α)/c-Jun N-terminal kinase (JNK) signaling pathway. PC12 cells were divided into a Control group (no treatment), Model group (treated with CoCl2 for 24 h), negative control (NC) group (transfected with miR-374a negative control sequence and treated with CoCl2 for 24 h), and miR-374a mimic group (transfected with miR-374a mimics and treated with CoCl2 for 24 h). The viability and apoptosis of PC12 cells were determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and flow cytometry, while the mitochondrial membrane potential (MMP) and reactive oxygen species (ROS) content were assessed by Rh123 and dichloro-dihydro-fluorescein diacetate (DCFH-DA) methods. The expression of miR-374a and GADD45α/JNK proteins was detected using quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and Western blot. A significant decrease was found in the survival rate, MMP and miR-374a expression, while an increase was shown in the ROS content and GADD45α and p-JNK expression in hypoxic PC12 cells (all P < 0.05). A luciferase reporter gene assay demonstrated that GADD45α is the target gene of miR-374a. When transfected with miR-374a mimics, hypoxic PC12 cells showed an obvious elevation in survival rate and MMP but a great reduction in cell apoptosis and ROS content, as well as in the expression of GADD45α and p-JNK proteins (all P < 0.05). MiR-374a can protect PC12 cells against hypoxia-induced injury by inhibiting the GADD45α/JNK pathway, enhancing cell viability, suppressing oxidative stress, and inhibiting cell apoptosis, thereby becoming a potential therapeutic target for hypoxic damage.

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Acknowledgements

The authors appreciate the reviewers for their useful comments in this paper.

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This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

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  1. Department of Neurological Rehabilitation, Beijing Rehabilitation Hospital, Capital Medical University, Xixiazhuang, Badachu Road, Shijingshan District, Beijing, 100144, China

    Weijun Gong, Shuyan Qie & Jianing Xi

  2. Beijing Rehabilitation Medicine Academy, Capital Medical University, Beijing, 100069, China

    Peiling Huang

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  1. Weijun Gong
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  2. Shuyan Qie
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Correspondence to Jianing Xi.

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Gong, W., Qie, S., Huang, P. et al. Protective Effect of miR-374a on Chemical Hypoxia-Induced Damage of PC12 Cells In Vitro via the GADD45α/JNK Signaling Pathway. Neurochem Res 43, 581–590 (2018). https://doi.org/10.1007/s11064-017-2452-0

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  • DOI: https://doi.org/10.1007/s11064-017-2452-0

Keywords

  • MiR-374a
  • GADD45α/JNK signaling pathway
  • Hypoxia
  • PC12 cells