Regulation of Prdx6 by Nrf2 Mediated Through aiPLA2 in White Matter Reperfusion Injury
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Regulation of Prdx6 by Nrf2 Mediated Through aiPLA2 in White Matter Reperfusion Injury Amita Daverey 1
&
Sandeep K. Agrawal 1
Received: 17 July 2020 / Accepted: 18 October 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
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. Keywords Prdx6 . White matter . Hypoxia . Reperfusion . Spinal cord injury . Neuroprotection
Abbreviations aiPLA2 ARE cDNA DAPI DC FBS GFAP H and E IACUC KEAP1 MAP 2
MJ33 Ca2+-independent phospholipase A2 Antioxidant response elements Complementary DNA 4′,6-Diamidino-2-phenylindole Dorsal column Fetal bovine serum Glial fibrillary acidic protein Hematoxylin and eosin Institutional Animal Care and Use Committee Kelch-like ECH-associated protein 1 Microtubule-associated protein 2
Supplementary Information The online version contains supplementary material available at https://doi.org/10.1007/s12035-020-02182-z. * Amita Daverey [email protected] Sandeep K. Agrawal [email protected] 1
Department of Neurosurgery, University of Nebraska Medical Center, Omaha, NE 68198-7690, USA
NOX2 NF-200 Nrf2 PBS-T Prdx6 PVDF qRT-PCR RIPA ROS RT SDS-PAGE SCI TBI TLR4 TUNEL UNMC WC
1-Hexadecyl-3-(trifluoroethyl)-sn-glycero-2phosphomethanol lithium NADPH oxidase 2 Neurofilament 200 Nuclear factor erythroid 2–related factor 2 Phosphate-buffered saline containing 0.1% Tween-20 Peroxiredoxin-6 Polyvinylidene difluoride Quantitative real-time PCR Radioimmunoprecipitation R
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