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Hypericum perforatum Attenuates Spinal Cord Injury-Induced Oxidative Stress and Apoptosis in the Dorsal Root Ganglion of Rats: Involvement of TRPM2 and TRPV1 Channels

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Abstract

Oxidative stress and cytosolic Ca2+ overload have important roles on apoptosis in dorsal root ganglion (DRG) neurons after spinal cord injury (SCI). Hypericum perforatum (HP) has an antioxidant property in the DRGs due to its ability to modulate NADPH oxidase and protein kinase C pathways. We aimed to investigate the protective property of HP on oxidative stress, apoptosis, and Ca2+ entry through transient receptor potential melastatin 2 (TRPM2) and transient receptor potential vanilloid 1 (TRPV1) channels in SCI-induced DRG neurons of rats. Rats were divided into four groups as control, HP, SCI, and SCI + HP. The HP groups received 30 mg/kg HP for three concessive days after SCI induction. The SCI-induced TRPM2 and TRPV1 currents and cytosolic free Ca2+ concentration were reduced by HP. The SCI-induced decrease in glutathione peroxidase and cell viability values were ameliorated by HP treatment, and the SCI-induced increase in apoptosis, caspase 3, caspase 9, cytosolic reactive oxygen species (ROS) production, and mitochondrial membrane depolarization values in DRG of SCI group were overcome by HP treatment. In conclusion, we observed a protective role of HP on SCI-induced oxidative stress, apoptosis, and Ca2+ entry through TRPM2 and TRPV1 in the DRG neurons. Our findings may be relevant to the etiology and treatment of SCI by HP.

Possible molecular pathways of involvement of Hypericum perforatum (HP) on apoptosis, oxidative stress, and calcium accumulation through TRPM2 and TRPV1 channels in DRG neurons of SCI-induced rats. The TRPM2 channel is activated by ADP-ribose and oxidative stress through activation of ADP-ribose pyrophosphate although it was inhibited by N-(p-amylcinnamoyl) anthranilic acid (ACA) and 2-aminoethyl diphenylborinate (2APB). The TRPV1 channel is activated by oxidative stress and capsaicin and it is blocked by capsazepine. Injury in the DRG can result in augmented ROS release, leading to Ca2+ uptake through TRPM2 and TRPV1 channels. Mitochondria were reported to accumulate Ca2+, provided intracellular Ca2+ rises, thereby leading to depolarization of mitochondrial membranes and release of apoptosis-inducing factors such as caspase 3 and caspase 9. HP via regulation of NADPH oxidase and PKC inhibits TRPM2 and TRPV1 channels. The molecular pathway may be a cause of SCI-induced pain and neuronal death, and the subject should be urgently investigated.

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Abbreviations

[Ca2+]c :

Intracellular free calcium ion concentration

2-APB:

2-Aminoethyl diphenylborinate

ACA:

N-(p-Amylcinnamoyl) anthranilic acid

CAP:

Capsaicin

CPZ:

Capsazepine

DRG:

Dorsal root ganglion

HP:

Hypericum perforatum

LP:

Lipid peroxidation

ROS:

Reactive oxygen species

SCI:

Spinal cord injury

TRP:

Transient receptor potential

TRPM2:

Transient receptor potential melastatin 2

TRPV1:

Transient receptor potential vanilloid 1

WC:

Whole cell

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Acknowledgments

The authors thank Dr. Peter J Butterworth (King’s College, London, UK) for helpful discussions on the manuscript. In addition, the authors thank Muhammet Şahin, technician Fatih Şahin, and research assistant Bilal Çiğ for helping with antioxidant analysis, patch-clamp analyses, and graphical abstract revision.

Conflict of Interest

The authors declare that there is no conflict of interest in the current study.

Financial Disclosure

The study was partially supported by the Unit of Scientific Research Project, Suleyman Demirel University (NO: BAP: 3266-TU2-12). There is no financial disclosure for the current study.

Author Contributions

MN and NŞ formulated the present hypothesis and were responsible for writing the report. ÜSÖ was responsible for induction of SCI. VG was responsible for DRG isolation and cytosolic Ca2+ release analyses.

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Authors and Affiliations

  1. Department of Neurosurgery, Faculty of Medicine, Suleyman Demirel University, Isparta, Turkey

    Ümit Sinan Özdemir & Nilgün Şenol

  2. Neuroscience Research Center, Suleyman Demirel University, TR-32260, Isparta, Turkey

    Mustafa Nazıroğlu & Vahid Ghazizadeh

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  1. Ümit Sinan Özdemir
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  2. Mustafa Nazıroğlu
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  3. Nilgün Şenol
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  4. Vahid Ghazizadeh
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Correspondence to Mustafa Nazıroğlu.

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Özdemir, Ü.S., Nazıroğlu, M., Şenol, N. et al. Hypericum perforatum Attenuates Spinal Cord Injury-Induced Oxidative Stress and Apoptosis in the Dorsal Root Ganglion of Rats: Involvement of TRPM2 and TRPV1 Channels. Mol Neurobiol 53, 3540–3551 (2016). https://doi.org/10.1007/s12035-015-9292-1

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