2. Academic Validation
  3. Butylphthalide inhibits ferroptosis and ameliorates cerebral Ischaemia-Reperfusion injury in rats by activating the Nrf2/HO-1 signalling pathway

Butylphthalide inhibits ferroptosis and ameliorates cerebral Ischaemia-Reperfusion injury in rats by activating the Nrf2/HO-1 signalling pathway

  • Neurotherapeutics. 2024 Sep;21(5):e00444. doi: 10.1016/j.neurot.2024.e00444.
Meilin Sun 1,  Junmin Chen 2,  Fan Liu 2,  Pei Li 3,  Jundong Lu 4,  Shihao Ge 2,  Lele Wang 2,  Xin Zhang 2,  Xiaopeng Wang 5
Affiliations

Affiliations

  • 1 Department of Neurology, The Second Hospital of Hebei Medical University, Shijiazhuang 050000, Hebei, China; Department of Neurology, Xingtai People's Hospital, Xingtai 054001, Hebei, China.
  • 2 Department of Neurology, The Second Hospital of Hebei Medical University, Shijiazhuang 050000, Hebei, China.
  • 3 Department of Neurology, Tangshan Gongren Hospital, Tangshan 063000, Hebei, China.
  • 4 Department of Neurology, Baoding First Central Hospital, Baoding 071000, Hebei, China.
  • 5 Department of Neurology, The Second Hospital of Hebei Medical University, Shijiazhuang 050000, Hebei, China. Electronic address: wangxiaopeng3035@163.com.
PMID: 39353831 DOI: 10.1016/j.neurot.2024.e00444
Abstract

This study aims to investigate whether butylphthalide can inhibit Ferroptosis and ameliorate cerebral ischaemia-reperfusion (I/R) injury in rats by activating the nuclear factor erythroid 2-related factor 2 (Nrf2) / heme oxygenase-1 (HO-1) signalling pathway, known for its antioxidative and cytoprotective properties. Middle cerebral artery occlusion reperfusion (MCAO/R) rat models were established. Male rats were randomly divided into five groups: a sham-operated group (sham), MCAO/R group, MCAO/R ​+ ​ML385 (Nrf2-specific inhibitor) group, MCAO/R ​+ ​NBP (butylphthalide) group and MCAO/R ​+ ​ML385 ​+ ​NBP group. The effect of butylphthalide on cerebral I/R injury was evaluated using neurological deficit scores. The expression levels of Nrf2, HO-1, Glutathione Peroxidase 4 (GPX4), acyl-CoA synthetase long-chain family member 4 (ACSL4) and Transferrin Receptor 1 (TfR1) protein were detected using Western blot. Moreover, the expression levels of GPX4, HO-1 and TfR1 mRNA were determined through real-time fluorescence quantitative reverse transcription polymerase chain reaction. The distribution of Nrf2, HO-1, GPX4 and TfR1 was detected using immunohistochemical staining. The levels of iron and related lipid peroxidation indexes, such as reduced glutathione, Reactive Oxygen Species, malondialdehyde and nitric oxide, were measured using a kit. The changes in mitochondria were observed through transmission electron microscopy. Butylphthalide treatment significantly improved neurological dysfunction, reduced cerebral infarction volume and mitigated histopathological damage in MCAO/R rats. It induced the nuclear translocation of Nrf2 and upregulated HO-1 expression, which was attenuated by ML385. Butylphthalide also attenuated lipid peroxidation, iron accumulation and mitochondrial damage induced by MCAO/R. The expression of GPX4, ACSL4 and TfR1 proteins, as well as their mRNA levels, was modulated through butylphthalide treatment, with improvements observed in mitochondrial morphology. Butylphthalide exerts neuroprotective effects by attenuating neurological dysfunction and Ferroptosis in MCAO/R rats through the activation of the Nrf2/HO-1 pathway and inhibition of lipid peroxidation and iron accumulation.

Keywords

Butylphthalide; Cerebral ischaemia reperfusion; Ferroptosis; Glutathione peroxidase 4; Heme oxygenase-1; Nuclear factor erythroid 2-related factor 2.

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