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Upregulation of miR-133a-3p in the Sciatic Nerve Contributes to Neuropathic Pain Development

Molecular Neurobiology volume 57, pages 3931–3942 (2020)Cite this article

Abstract

The micro (mi)RNAs expressed in the sciatic nerve of streptozotocin (STZ)-induced diabetic rats were evaluated in terms of their therapeutic potential in patients with diabetic neuropathic pain (DNP). Relative miRNA expression in sciatic nerve with DNP was analyzed using next-generation sequencing and quantitative PCR. Potential downstream targets of miRNAs were predicted using Ingenuity Pathway Analysis and the TargetScan database. In vitro experiments were performed using miR-133a-3p-transfected RSC96 Schwann cells. We performed micro-Western and Western blotting and immunofluorescence analyses to verify the role of miR-133a-3p. In vivo, the association between miR-133a-3p with DNP was analyzed via AAV-miR-133a-3p intraneural (intra-epineural but extrafascicular) injection into the sciatic nerve of normal rats or injection of an miR-133a-3p antagomir into the sciatic nerve of diabetes mellitus (DM) rats. miR-133a-3p mimics transfected into RSC96 Schwann cells increased VEGFR-2, p38α MAPK, TRAF-6, and PIAS3 expression and reduced NFκB p50 and MKP3 expression. In normal rats, AAV-miR-133a-3p delivery via intraneural injection into the sciatic nerve induced mechanical allodynia and p-p38 MAPK activation. In DM rats, miR-133a-3p antagomir administration alleviated DNP and downregulated p-p38 phosphorylation. Overexpression of miR-133a-3p in the sciatic nerve induced such pain. We suggest that miR-133a-3p is a potential therapeutic target for DNP.

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Abbreviations

miR-133a-3p:

microRNA-133a-3p

STZ:

streptozotocin

DNP:

diabetic neuropathic pain

VEGFR-2:

vascular endothelial growth factor receptor 2

P38α MAPK:

P38α mitogen-activated protein kinases

TRAF-6:

TNF receptor-associated factor-6

PIAS3:

E3 SUMO-protein ligase PIAS3

NFκB p50:

nuclear factor-κB p50

MKP3:

MAP kinase phosphatase 3

p-p38 MAPK:

phospho-p38 MAPK

DN:

diabetic neuropathy

DRG:

dorsal root ganglion

SD rats:

Sprague Dawley rats

AAV:

adeno-associated virus

NGS:

next-generation sequencing

RT-PCR:

reverse transcription PCR

qRT-PCR:

real-time quantitative reverse transcription PCR

rAAV:

recombinant adeno-associated virus

eGFP:

enhanced green fluorescent protein

GFAP:

glial fibrillary acidic protein

SDH:

spinal dorsal horn

NADH:

nicotinamide adenine dinucleotide (reduced)

ERKs:

extracellular signal-regulated kinase

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Acknowledgments

We also acknowledge the support of the Center for Resources, Research and Development of Kaohsiung Medical University for use of the confocal laser scanning microscope.

Funding

This study was supported by grants from Ministry of Science and Technology (MOST 103-2314-B-037-042, MOST 104-2314-B-037-022-MY2, MOST 106-2314-B-037-032, MOST 107-2314-B-037-035-MY2).

Author information

Affiliations

  1. Department of Microbiology and Immunology, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

    Lin-Li Chang & Yi-Hsuan Wang

  2. Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

    Lin-Li Chang & Kuang-Yi Tseng

  3. Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

    Lin-Li Chang

  4. Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan

    Lin-Li Chang

  5. Department of Neurosurgery, Chang Gung Memorial Hospital-Kaohsiung Medical Center, Chang Gung University College of Medicine, Kaohsiung, Taiwan

    Hung-Chen Wang

  6. Department of Anesthesiology, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan

    Kuang-Yi Tseng, Miao-Pei Su & Kuang-I Cheng

  7. Division of Colorectal Surgery, Department of Surgery, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan

    Jaw-Yuan Wang

  8. Department of Surgery, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

    Jaw-Yuan Wang

  9. Center for Biomarkers and Biotech Drugs, Kaohsiung Medical University, Kaohsiung, Taiwan

    Jaw-Yuan Wang

  10. Physical Education Center, Kaohsiung Medical University, Kaohsiung, Taiwan

    Yi-Ta Chuang

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  1. Lin-Li Chang
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  2. Hung-Chen Wang
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Contributions

Lin-Li Chang was responsible for conceiving, organizing, and implementing the research protocol, interpreting the data, guiding discussions of the results, and drafting the manuscript. Hung-Chen Wang, Kuang-Yi Tseng, Jaw-Yuan Wang, Miao-Pei Su, Yi-Da Chuang, and Yi-Hsuan Wang participated in the experiment and data evaluation. Kuang-I Cheng coordinated the study, interpreted the data, supervised the research program, and contributed to the screening process. All authors read and approved the manuscript.

Corresponding author

Correspondence to Kuang-I Cheng.

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The authors declare that they have no competing interest. Additionally, all authors have read the journal’s authorship agreement and policy on disclosure of potential conflicts of interest. All animal experimental procedures were approved by the Kaohsiung Institutional Animal Care and Use Committee (approval no. 102162).

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Chang, LL., Wang, HC., Tseng, KY. et al. Upregulation of miR-133a-3p in the Sciatic Nerve Contributes to Neuropathic Pain Development. Mol Neurobiol 57, 3931–3942 (2020). https://doi.org/10.1007/s12035-020-01999-y

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  • DOI: https://doi.org/10.1007/s12035-020-01999-y

Keywords

  • Diabetic neuropathic pain
  • miR-133a-3p
  • p38 MAPK
  • Schwann cells