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Regulation of Prdx6 by Nrf2 Mediated Through aiPLA2 in White Matter Reperfusion Injury

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Abstract

Hypoxia and reperfusion produces overproduction of ROS (reactive oxygen species), which may lead to mitochondrial dysfunction leading to cell death and apoptosis. Here, we explore the hypothesis that Prdx6 protects the spinal cord white matter from hypoxia-reperfusion injury and elucidate the possible mechanism by which Prdx6 elicits its protective effects. Briefly, rats were deeply anesthetized with isoflurane. A 30-mm section of the spinal cord was rapidly removed and placed in cold Ringer’s solution (2–4 °C). The dissected dorsal column was exposed to hypoxia with 95% N2 and 5% CO2 and reperfusion with 95% O2 and 5% CO2. The expression of Prdx6 significantly upregulated in white matter after hypoxia compared to the sham group, whereas reperfusion caused a gradual decrease in Prdx6 expression after reperfusion injury. For the first time, our study revealed the novel expression and localized expression of Prdx6 in astrocytes after hypoxia, and possible communication of astrocytes and axons through Prdx6. The gradual increase in Nrf2 expression suggests a negative regulation of Prdx6 through Nrf2 signaling. Furthermore, inhibition of aiPLA2 activity of Prdx6 by MJ33 shows that the regulation of Prdx6 by Nrf2 is mediated through aiPLA2 activity. The present study uncovers a differential distribution of Prdx6 in axons and astrocytes and regulation of Prdx6 in hypoxia-reperfusion injury. The low levels of Prdx6 in reperfusion injury lead to increased inflammation and apoptosis in the white matter; therefore, the results of this study suggest that Prdx6 has a protective role in spinal hypoxia-reperfusion injury.

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Data Availability

The authors confirm that the data supporting the findings of this study are available within the article and its Supplementary material.

Abbreviations

aiPLA2:

Ca2+-independent phospholipase A2

ARE:

Antioxidant response elements

cDNA:

Complementary DNA

DAPI:

4′,6-Diamidino-2-phenylindole

DC:

Dorsal column

FBS:

Fetal bovine serum

GFAP:

Glial fibrillary acidic protein

H and E:

Hematoxylin and eosin

IACUC:

Institutional Animal Care and Use Committee

KEAP1:

Kelch-like ECH-associated protein 1

MAP 2:

Microtubule-associated protein 2

MJ33:

1-Hexadecyl-3-(trifluoroethyl)-sn-glycero-2-phosphomethanol lithium

NOX2:

NADPH oxidase 2

NF-200:

Neurofilament 200

Nrf2:

Nuclear factor erythroid 2–related factor 2

PBS-T:

Phosphate-buffered saline containing 0.1% Tween-20

Prdx6:

Peroxiredoxin-6

PVDF:

Polyvinylidene difluoride

qRT-PCR:

Quantitative real-time PCR

RIPA:

Radioimmunoprecipitation

ROS:

Reactive oxygen species

RT:

Room temperature

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

SCI:

Spinal cord injury

TBI:

Traumatic brain injury

TLR4:

Toll-like receptor 4

TUNEL:

Terminal deoxynucleotidyl transferase dUTP nick end labeling

UNMC:

University of Nebraska Medical Center

WC:

Whole spinal cord

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Acknowledgments

The authors would like to thank the Department of Neurosurgery, University of Nebraska Medical Center for supporting this research.

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Authors and Affiliations

  1. Department of Neurosurgery, University of Nebraska Medical Center, Omaha, NE, 68198-7690, USA

    Amita Daverey & Sandeep K. Agrawal

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  1. Amita Daverey
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Contributions

AD designed and performed experiments, analyzed and interpreted data, and wrote and revised the manuscript. SA conceived, designed and supervised the project, performed some experiments, analyzed data, and was responsible for data interpretation, and writing and revising the manuscript. All authors read and approved the final manuscript.

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Correspondence to Amita Daverey.

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The animal experiments were carried out in accordance with the UNMC (University of Nebraska Medical Center) Occupational Health and Safety Program for Animal Contact and all the animal procedures used in this study were approved by the IACUC (Institutional Animal Care and Use Committee) of the UNMC.

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The authors declare that they have no competing interests.

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Daverey, A., Agrawal, S.K. Regulation of Prdx6 by Nrf2 Mediated Through aiPLA2 in White Matter Reperfusion Injury. Mol Neurobiol 58, 1275–1289 (2021). https://doi.org/10.1007/s12035-020-02182-z

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