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Over-expression of miR-3584-5p Represses Nav1.8 Channel Aggravating Neuropathic Pain caused by Chronic Constriction Injury

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Abstract

Nav1.8, a tetrodotoxin-resistant voltage-gated sodium channels (VGSCs) subtype encoded by SCN10A, which plays an important role in the production and transmission of peripheral neuropathic pain signals. Studies have shown that VGSCs may be key targets of MicroRNAs (miRNAs) in the regulation of neuropathic pain. In our study, bioinformatics analysis showed that the targeting relationship between miR-3584-5p and Nav1.8 was the most closely. The purpose of this study was to investigate the roles of miR-3584-5p and Nav1.8 in neuropathic pain. The effects of miR-3584-5p on chronic constriction injury (CCI)-induced neuropathic pain in rats was investigated by intrathecal injection of miR-3584-5p agomir (an agonist, 20 μM, 15 μL) or antagomir (an antagonist, 20 μM, 15 μL). The results showed that over-expression of miR-3584-5p aggravated neuronal injury by hematoxylin–eosin (H&E) staining and mechanical/thermal hypersensitivity in CCI rats. MiR-3584-5p indirectly inhibited the expression of Nav1.8 by up-regulating the expression of key proteins in the ERK5/CREB signaling pathway, and also inhibited the current density of the Nav1.8 channel, changed its channel dynamics characteristic, thereby accelerating the transmission of pain signals, and further aggravating pain. Similarly, in PC12 and SH-SY5Y cell cultures, miR-3584-5p increased the level of reactive oxygen species (ROS) and inhibited mitochondrial membrane potential (Δψm) in the mitochondrial pathway, decreased the ratio of apoptosis-related factor Bcl-2/Bax, and thus promoted neuronal apoptosis. In brief, over-expression of miR-3584-5p aggravates neuropathic pain by directly inhibiting the current density of Nav1.8 channel and altering its channel dynamics, or indirectly inhibiting Nav1.8 expression through ERK5/CREB pathway, and promoting apoptosis through mitochondrial pathway.

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Data Availability

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

Abbreviations

MiRNAs:

MicroRNAs

CCI:

Chronic constriction injury

qRT-PCR:

Quantitative real-time polymerase chain reaction

H&E:

Hematoxylin-eosin

DRG:

Dorsal root ganglion

ERK5:

Extracellular signal regulated kinase 5

CREB:

CAMP-response element binding protein

ROS:

Reactive oxygen species

Bcl-2:

B-cell-lymphoma-2

Bax:

Bcl2-associated X-protein

AO/EB:

Acridine orange and ethidium bromide

Δψm:

Mitochondrial membrane potential

VGSCs:

Voltage-gated sodium channels

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Acknowledgements

This study was funded by the National Natural Science Foundation of Liaoning Province (grant number: 20180550253); Scientific Research Foundation of the Education Department of Liaoning Province (grant number LJKZ0956); Scientific Research Staring Foundation for the Returned Overseas Scholars, Shenyang Pharmaceutical University (grant number GGJJ2021101); National Science and Technology Major Project of the Ministry of Science and Technology of China (grant number 2018ZX09735005).

Funding

This work was supported by the National Natural Science Foundation of Liaoning Province (grant number: 20180550253); Scientific Research Foundation of the Education Department of Liaoning Province (grant number LJKZ0956); Scientific Research Staring Foundation for the Returned Overseas Scholars, Shenyang Pharmaceutical University (grant number GGJJ2021101); National Science and Technology Major Project of the Ministry of Science and Technology of China (grant number 2018ZX09735005).

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Authors and Affiliations

  1. Department of Pharmacology, Shenyang Pharmaceutical University, Shenyang, 110016, Liaoning, People’s Republic of China

    Ran Yang, Qian-qian Wang, Yuan Feng, Xue-hao Li, Gui-xia Li, Feng-lin She, Xi-jin Zhu & Chun-li Li

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  1. Ran Yang
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Contributions

All authors contributed to the study conception and design. Validation, formal analysis, data curation, writing-original draft, writing-Review & editing and visualization were performed by Ran Yang. Conceptualization, methodology, formal analysis, investigation and data curation were performed by Qian-qian Wang, Yuan Feng and Xue-hao Li. Conceptualization, methodology and formal analysis were performed by Gui-xia Li. Methodology, formal analysis and data curation were performed by Xi-jin Zhu and Feng-lin She. Conceptualization, resources, writing-original draft, writing-review & editing, supervision, project administration and Funding acquisition were performed by Chun-li Li. All authors read and approved the final manuscript.

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Correspondence to Chun-li Li.

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Yang, R., Wang, Qq., Feng, Y. et al. Over-expression of miR-3584-5p Represses Nav1.8 Channel Aggravating Neuropathic Pain caused by Chronic Constriction Injury. Mol Neurobiol 60, 5237–5255 (2023). https://doi.org/10.1007/s12035-023-03394-9

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