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Network Pharmacology Prediction and Experimental Verification for Anti-Ferroptosis of Edaravone After Experimental Intracerebral Hemorrhage

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Abstract

Neuronal ferroptosis plays an important role in secondary brain injuries after intracerebral hemorrhage (ICH). Edaravone (Eda) is a promising free radical scavenger that inhibits ferroptosis in neurological diseases. However, its protective effects and underlying mechanisms in ameliorating post-ICH ferroptosis remain unclear. We employed a network pharmacology approach to determine the core targets of Eda against ICH. Forty-two rats were subjected to successful striatal autologous whole blood injection (n=28) or sham operation (n=14). The 28 blood-injected rats were randomly assigned to either the Eda or vehicle group (n=14) for immediate administration and then for 3 consecutive days. Hemin-induced HT22 cells were used for in vitro studies. The effects of Eda in ICH on ferroptosis and the MEK/ERK pathway were investigated in vivo and in vitro. Network pharmacology-based analysis revealed that candidate targets of Eda-treated ICH might be related to ferroptosis; among which prostaglandin G/H synthase 2 (PTGS2) was a ferroptosis marker. In vivo experiments showed that Eda alleviated sensorimotor deficits and decreased PTGS2 expression (all p<0.05) after ICH. Eda rescued neuron pathological changes after ICH (increased NeuN+ cells and decreased FJC+ cells, all p<0.01). In vitro experiments showed that Eda reduced intracellular reactive oxygen species and reversed mitochondria damage. Eda repressed ferroptosis by decreasing malondialdehyde and iron deposition and by influencing ferroptosis-related protein expression (all p<0.05) in ICH rats and hemin-induced HT22 cells. Mechanically, Eda significantly suppressed phosphorylated-MEK and phosphorylated-ERK1/2 expression. These results indicate that Eda has protective effects on ICH injury through ferroptosis and MEK/ERK pathway suppression.

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Data Availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

Abbreviations

ALB :

albumin

COX2 :

cyclooxygenase-2

ERK :

extracellular-regulated kinase

FJC :

Fluoro-Jade C

GPX4 :

glutathione peroxidase 4

ICH :

intracerebral hemorrhage

KEGG :

Kyoto Encyclopedia of Genes and Genomes

PPI :

protein-protein interaction

PTGS2 :

prostaglandin G/H synthase 2

ROS :

reactive oxygen species

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Funding

This study was supported by grants from the Natural Science Foundation of China (Key Program: 82130035; General Program: 81771137, 81971103), the Scientific and Technical Project of Guangdong Province (2017A030303011), Sun Yat-sen University Clinical Research 5010 Program (2018001), the Guangdong Provincial Key Laboratory for Diagnosis and Treatment of Major Neurological Diseases (2017B030314103), Guangdong Provincial Clinical Research Center for Neurological Diseases (2020B1111170002), the Southern China International Cooperation Base for Early Intervention and Functional Rehabilitation of Neurological Diseases (2015B050501003), Guangdong Provincial Engineering Center for Major Neurological Disease Treatment, and Guangdong Provincial Translational Medicine Innovation Platform for Diagnosis and Treatment of Major Neurological Disease.

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  1. Department of Neurology, The First Affiliated Hospital, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Diagnosis and Treatment of Major Neurological Diseases, National Key Clinical Department of Key Discipline of Neurology, Southern China International Cooperation Base for Early Intervention and Functional Rehabilitation of Neurological Diseases, No.58 Zhongshan Road 2, Guangzhou, 510080, China

    Suhan Cao, Jiating Wei, Yuangui Cai, Zhiyi Xiong, Jianle Li, Zimu Jiang, Xueling Zhou, Baozi Huang & Jinsheng Zeng

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Correspondence to Jinsheng Zeng.

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All experimental procedures were approved by the Institutional Animal Ethical Committee of Sun Yat-Sen University (Approval No: SYSU-IACUC-2022-001631) and were conducted following the “Guide for the Care and Use of Laboratory Animals” by the National Institute of Health in China.

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Cao, S., Wei, J., Cai, Y. et al. Network Pharmacology Prediction and Experimental Verification for Anti-Ferroptosis of Edaravone After Experimental Intracerebral Hemorrhage. Mol Neurobiol 60, 3633–3649 (2023). https://doi.org/10.1007/s12035-023-03279-x

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