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Minocycline alleviates nociceptive response through modulating the expression of NR2B subunit of NMDA receptor in spinal cord of rat model of painful diabetic neuropathy

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Abstract

Background

It has been reported that neuropathic pain can be overcome by targeting the NR2B subunit of N-methyl-D-aspartate receptors (NR2B). This study aimed to investigate the effects of minocycline on phosphorylated and total expression of NR2B in the spinal cord of rats with diabetic neuropathic pain.

Methods

A total of 32 Sprague–Dawley male rats were randomly assigned into four groups (n = 8); control healthy, control diabetic (PDN), and PDN rats that received 80 µg or 160 µg intrathecal minocycline respectively. The rats were induced to develop diabetes and allowed to develop into the early phase of PDN for two weeks. Hot-plate and formalin tests were conducted. Intrathecal treatment of minocycline or normal saline was conducted for 7 days. The rats were sacrificed to obtain the lumbar enlargement region of the spinal cord (L4-L5) for immunohistochemistry and western blot analyses to determine the expression of phosphorylated (pNR2B) and total NR2B (NR2B).

Results

PDN rats showed enhanced flinching (phase 1: p < 0.001, early phase 2: p < 0.001, and late phase 2: p < 0.05) and licking responses (phase 1: p < 0.001 and early phase 2: p < 0.05). PDN rats were also associated with higher spinal expressions of pNR2B and NR2B (p < 0.001) but no significant effect on thermal hyperalgesia. Minocycline inhibited formalin-induced flinching and licking responses (phase 1: p < 0.001, early phase 2: p < 0.001, and late phase 2: p < 0.05) in PDN rats with lowered spinal expressions of pNR2B (p < 0.01) and NR2B (p < 0.001) in a dose-dependent manner.

Conclusion

Minocycline alleviates nociceptive responses in PDN rats, possibly via suppression of NR2B activation. Therefore, minocycline could be one of the potential therapeutic antinociceptive drugs for the management of neuropathic pain.

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Data Availability

All the data are shown in the manuscript.

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Acknowledgements

We would like to thank Universiti Sains Malaysia and Ministry of Higher Education, Malaysia for the funding provided; Research University grant numbered RUI 1001/PPSK/812139 and Fundamental Research Grant Scheme numbered 203/PPSK/6171189. We also thank the staffs of Physiology Laboratory, Animal Research and Service Center, Central Research Laboratory, Cranial and Maxillofacial and Molecular and Molecular Laboratory for the direct and indirect technical guides.

Funding

This study was funded under Research University (RUI) grant (1001/PPSK/812139) by Universiti Sains Malaysia and Fundamental Research Grant Scheme (FRGS) (203/PPSK/6171189) by Ministry of Higher Education, Malaysia.

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Authors and Affiliations

  1. Department of Physiology, Universiti Sains Malaysia Health Campus, 16150 Kubang Kerian, Kelantan, Malaysia

    Che Aishah Nazariah Ismail & Che Badariah Abd Aziz

  2. Brain and Behaviour Cluster, Universiti Sains Malaysia Health Campus, 16150 Kubang Kerian, Kelantan, Malaysia

    Che Aishah Nazariah Ismail

  3. Biostatistics and Research Methodology Unit, School of Medical Sciences, Universiti Sains Malaysia Health Campus, 16150 Kubang Kerian, Kelantan, Malaysia

    Anis Kausar Ghazali

  4. School of Health Sciences, Universiti Sains Malaysia Health Campus, 16150 Kubang Kerian, Kelantan, Malaysia

    Rapeah Suppian & Idris Long

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Contributions

Ismail CAN is responsible for the whole manuscript including figures and table. Ghazali AK guides on statistical analyses of the experimental data. Suppian R and Ab Aziz CB are responsible for proofreading and improving the quality of the manuscript. Long I provides research funding and finalizes the manuscript.

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Correspondence to Che Aishah Nazariah Ismail.

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All the experimentation of the study was approved by the Animal Ethics Committee, Universiti Sains Malaysia [USM/Animal Ethics Approval/2014 (91) (560)].

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Ismail, C.A.N., Ghazali, A.K., Suppian, R. et al. Minocycline alleviates nociceptive response through modulating the expression of NR2B subunit of NMDA receptor in spinal cord of rat model of painful diabetic neuropathy. J Diabetes Metab Disord 20, 793–803 (2021). https://doi.org/10.1007/s40200-021-00820-4

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