Work

Molecular hydrogen exerts neuroprotection against cerebral ischemia–reperfusion injury via inhibition of Nrf2/HO-1-dependent ferroptosis in mice

Year: 2026

Type: article

Abstract: Background: Cerebral ischemia-reperfusion injury (CIRI) represents secondary brain damage induced by reperfusion after ischemic insult. Molecular hydrogen(H₂)is a highly permeable, biosafe and selective antioxidant, which has emerged as a promising redox-modulating agent in various oxidative stress-related disorders. Clinically, a balanced gas mixture composed of 67% (v/v) hydrogen and 33% (v/v) oxygen is a standardized, widely accepted inhalation formulation for molecular hydrogen delivery. This rational oxygen supplementation avoids hypoxia caused by pure high-concentration hydrogen inhalation and ensures clinical safety. However, the precise mechanism by which molecular hydrogen regulates Nrf2/HO-1 signaling and neuronal ferroptosis in cerebral I/R injury remains poorly clarified. Methods: In vivo experiments were performed using C57BL/6 mice subjected to middle cerebral artery occlusion/reperfusion (MCAO/R), while in vitro studies used SH‑SY5Y cells exposed to oxygen‑glucose deprivation/reoxygenation (OGD/R). Infarct volume was assessed by TTC staining, and histopathological changes by HE and Nissl staining. Protein levels of Nrf2‑mediated ferroptosis‑related markers were detected by Western blot and immunohistochemistry, and iron transport‑related genes by qPCR. Oxidative stress, mitochondrial membrane potential (MMP), and apoptosis were measured via biochemical assays and flow cytometry. ML385 (Nrf2 inhibitor) and Ferrostatin‑1 (ferroptosis inhibitor) were used for mechanistic validation. Results: Inhalation of 66.7% H₂/33.3% O₂ markedly reduced cerebral infarction, neurological deficits, histopathological injury, and ferroptosis in MCAO/R mice. In SH‑SY5Y cells, H₂ improved cell viability and MMP, and reduced apoptosis and ferroptosis following OGD/R. H₂ upregulated Nrf2, GPX4, and HO‑1 expression, similar to Ferrostatin‑1, whereas ML385 abolished these beneficial effects. Conclusion: This study is the first to demonstrate that H₂ protects against CIRI by attenuating oxidative stress and ferroptosis via the Nrf2/GPX4/HO‑1 pathway, revealing a novel mechanism underlying hydrogen‑mediated neuroprotection.

Source: Scientific Reports

DOI: 10.1038/s41598-026-67710-8

Authors: Zhen-kui Wang, Wei Feng, Jie Zhou, Lei Huang, Hong-sheng Chen, Li Xue

Institutions: Harbin Medical University, Fourth Affiliated Hospital of Harbin Medical University, Dali University

Language: English

Topic: Hydrogen's biological and therapeutic effects

Subfield: Surgery

Field: Medicine

Domain: Health Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $2,690