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Inhibition of Ferroptosis Alleviates Early Brain Injury After Subarachnoid Hemorrhage In Vitro and In Vivo via Reduction of Lipid Peroxidation

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Abstract

Subarachnoid hemorrhage (SAH) is a serious cerebrovascular disease with high mortality, and the mean age at morbidity is younger than in other types of stroke. Early brain injury (EBI) plays a key role in the poor prognoses of SAH. In EBI, multiple forms of cell death have been identified and well studied; however, the role of ferroptosis has not been elucidated. Hence, in this study, we developed an in vivo (SAH rat model) and in vitro model (SH-SY5Y oxyhemoglobin injury model) to understand the role of ferroptosis in EBI, then explored the protective mechanism of ferrostatin-1 (Fer-1). Firstly, we found that neurological scores, blood–brain barrier permeability, brain edema deteriorated after SAH in the in vivo model, cell viability was decreased after SAH in both cortex and SH-SY5Y cells. Further, iron content in cortex was increased after SAH, while transferrin receptor 1 and ferroportin (Fpn) were increased in oxyhemoglobin-treated in vitro model. Additionally, glutathione content and glutathione peroxidase 4 activity were reduced in SAH rats, and lipid peroxides were increased in the oxyhemoglobin-treated cells. Finally, administration of Fer-1 upregulated Fpn and decreased the iron content, then improved the lipid peroxidation and EBI. However, Fer-1 had no effect on the apoptosis. Our study indicated that the ferroptosis was involved in EBI of SAH, and the inhibitor Fer-1 provided neuroprotection against EBI by alleviating ferroptosis, the potential protective mechanism might be via suppressing lipid peroxidation.

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Abbreviations

BBB:

Blood–brain barrier

CCA:

Common carotid artery

DMT1:

Divalent metal transporter 1

DW:

Dry weight

EBI:

Early brain injury

ECA:

External carotid artery

Fer-1:

Ferrostatin-1

FJC:

Fluoro-Jade C

Fpn:

Ferroportin

GPx4:

Glutathione peroxidase 4

GSH:

Glutathione

Hb:

Oxyhemoglobin

ICH:

Intracranial hematoma

ICA:

Internal carotid artery

IRP1/2:

Iron regulatory proteins 1/2

OD:

Optical density

PBS:

Phosphate buffer solution

ROS:

Reactive oxygen species

SAH:

Subarachnoid hemorrhage

TEM:

Transmission electron microscope

TfR1:

Transferrin receptor 1

WW:

Wet weight

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Funding

This work is supported by the American Heart Association, USA (16SDG26420078), Natural Science Foundation of Heilongjiang Province, China (ZD2018018) and Innovation Fund of Harbin Medical University, China (YTSKYCX2018-38HYD).

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Author notes

  1. Ji Bihl and Huaizhang Shi have equally contributed to this work.

Authors and Affiliations

  1. Department of Neurosurgery, The First Affiliated Hospital, Harbin Medical University, Harbin, 150000, Heilongjiang, China

    Yuchen Li, Yao Liu, Pei Wu, Yang Tian, Binbing Liu & Huaizhang Shi

  2. Department of Pharmacology & Toxicology, Boonshoft School of Medicine, Wright State University, Dayton, OH, 45435, USA

    Yuchen Li, **ju Wang & Ji Bihl

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  1. Yuchen Li
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  2. Yao Liu
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Li, Y., Liu, Y., Wu, P. et al. Inhibition of Ferroptosis Alleviates Early Brain Injury After Subarachnoid Hemorrhage In Vitro and In Vivo via Reduction of Lipid Peroxidation. Cell Mol Neurobiol 41, 263–278 (2021). https://doi.org/10.1007/s10571-020-00850-1

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