Loureirin C inhibits ferroptosis after cerebral ischemia reperfusion through regulation of the Nrf2 pathway in mice

Yeshu Liu; Yan Mi; Yongping Wang; Qingqi Meng; Libin Xu; Yueyang Liu; Di Zhou; Yingjie Wang; Dong Liang; Wei Li; Ning Li; Yue Hou

doi:10.1016/j.phymed.2023.154729

Loureirin C inhibits ferroptosis after cerebral ischemia reperfusion through regulation of the Nrf2 pathway in mice

Phytomedicine. 2023 May:113:154729. doi: 10.1016/j.phymed.2023.154729. Epub 2023 Feb 26.

Authors

Yeshu Liu¹, Yan Mi¹, Yongping Wang², Qingqi Meng², Libin Xu², Yueyang Liu³, Di Zhou⁴, Yingjie Wang⁴, Dong Liang⁵, Wei Li⁶, Ning Li⁷, Yue Hou⁸

Affiliations

¹ Key Laboratory of Bioresource Research and Development of Liaoning Province, College of Life and Health Sciences, Northeastern University, Shenyang, China; National Frontiers Science Center for Industrial Intelligence and Systems Optimization, Key Laboratory of Data Analytics and Optimization for Smart Industry, Ministry of Education, Northeastern University, Shenyang, China.
² Key Laboratory of Bioresource Research and Development of Liaoning Province, College of Life and Health Sciences, Northeastern University, Shenyang, China.
³ Department of Pharmacology, Shenyang Pharmaceutical University, Shenyang, China.
⁴ School of Traditional Chinese Materia Medica, Key Laboratory for TCM Material Basis Study and Innovative Drug Development of Shenyang City, Shenyang Pharmaceutical University, Shenyang, China.
⁵ State Key Laboratory for Chemistry and Molecular Engineering of Medicinal Resources, School of Chemistry and Pharmaceutical Sciences, Guangxi Normal University, Guilin, China.
⁶ Faculty of Pharmaceutical Sciences, Toho University, Miyama 2-2-1, Funabashi, Chiba 274-8510, Japan.
⁷ School of Traditional Chinese Materia Medica, Key Laboratory for TCM Material Basis Study and Innovative Drug Development of Shenyang City, Shenyang Pharmaceutical University, Shenyang, China. Electronic address: 103050113@syphu.edu.cn.
⁸ Key Laboratory of Bioresource Research and Development of Liaoning Province, College of Life and Health Sciences, Northeastern University, Shenyang, China; National Frontiers Science Center for Industrial Intelligence and Systems Optimization, Key Laboratory of Data Analytics and Optimization for Smart Industry, Ministry of Education, Northeastern University, Shenyang, China. Electronic address: houyue@mail.neu.edu.cn.

PMID: 36878093
DOI: 10.1016/j.phymed.2023.154729

Abstract

Background: Ischemic stroke (IS) is considered as a serious cerebral vascular disease. Ferroptosis is a novel type of regulated cell death (RCD), that closely related to the occurrence and progress of IS. Loureirin C, a type of dihydrochalcone compound derived from the Chinese Dragon's blood (CDB). The effective components extracted from CDB have shown neuroprotective effects in ischemia reperfusion models. However, the role of Loureirin C in mice after IS is not well understood. Thus, it is worth to identify the effect and mechanism of Loureirin C on IS.

Purpose: The present research aims to prove the existence of ferroptosis in IS and explore whether Loureirin C can inhibit ferroptosis by regulating nuclear factor E2 related factor 2 (Nrf2) pathway in mice and exert neuroprotective effects on IS models.

Methods: Middle cerebral artery occlusion and reperfusion (MCAO/R) model was established to evaluate the occurrence of ferroptosis and the potential Loureirin C brain-protective effect in vivo. The analysis of free iron, glutamate content, reactive oxygen species (ROS) and lipid peroxidation levels, along with transmission electron microscope (TEM) was applied to prove the existence of ferroptosis. The function of Loureirin C on Nrf2 nuclear translocation was verified by immunofluorescence staining. In vitro, primary neurons and SH-SY5Y cells were processed with Loureirin C after oxygen and glucose deprivation-reperfusion (OGD/R). ELISA kits, western blotting, co-immunoprecipitation (Co-IP) analysis, immunofluorescence, and quantitative real-time PCR were devoted to proving the neuroprotective effects of Loureirin C on IS via regulating ferroptosis and Nrf2 pathways.

Results: The results showed that Loureirin C not only dramatically alleviated brain injury and inhibited neurons ferroptosis in mice after MCAO/R, but also dose-dependently reduce ROS accumulation in ferroptosis after OGD/R. Further, Loureirin C inhibits ferroptosis by activating Nrf2 pathway, and promoting nuclear translocation of Nrf2. Besides, Loureirin C increases heme oxygenase 1 (HO-1), quinone oxidoreductase 1 (NQO1) and glutathione peroxidase 4 (GPX4) content after IS. Intriguingly, the anti-ferroptosis effect of Loureirin C is weakened by Nrf2 knockdown.

Conclusion: Our discoveries first revealed that the inhibitory action of Loureirin C on ferroptosis may greatly depend on its adjusting effect on the Nrf2 pathway, suggesting that Loureirin C could act as a novel anti-ferroptosis candidate and play a therapeutic role in IS. These novel discoveries on the role of Loureirin C on IS models reveal an innovative method that may contribute to neuroprotection for the prevention of IS.

Keywords: Ferroptosis; Ischemic stroke (IS); Loureirin C; Nuclear factor E2 related factor 2 (Nrf2) pathway; Reactive oxygen species (ROS).

MeSH terms

Animals
Brain Ischemia* / drug therapy
Brain Ischemia* / metabolism
Humans
Infarction, Middle Cerebral Artery / drug therapy
Infarction, Middle Cerebral Artery / metabolism
Mice
NF-E2-Related Factor 2 / metabolism
Neuroblastoma* / drug therapy
Neuroprotective Agents* / pharmacology
Neuroprotective Agents* / therapeutic use
Oxidative Stress
Reactive Oxygen Species / metabolism
Reperfusion
Reperfusion Injury* / prevention & control
Signal Transduction

Substances

Reactive Oxygen Species
Neuroprotective Agents
NF-E2-Related Factor 2