Gastrodin alleviates hypobaric hypoxia-induced brain injury in rats by reducing neuronal ferroptosis via the P53/SLC7A11/GPX4 signaling axis

doi:10.12122/j.issn.1673-4254.2026.07.04

Abstract

Abstract:

Objective To investigate the neuroprotective effect of gastrodin (GAS) against hypobaric hypoxia (HH)-induced brain injury in rats and the underlying mechanism. Methods Twenty-four adult SD rats were randomized equally into normoxic control group, HH model group, low-dose (100 mg/kg) GAS group (HH+GAS-L group), and high-dose (200 mg/kg) GAS group (HH+GAS-H group). In the latter 3 groups, the rats were exposed to HH in a hypobaric oxygen chamber for 24 h to simulate the condition at an altitude of 6000 m, and GAS was administered intraperitoneally once daily for 7 days. Cerebral cortex tissues were collected for analysis of P53, SLC7A11, and GPX4 protein expressions using Western blotting and for determination of the levels of reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), and ferrous ion (Fe²⁺). In cultured HT22 neurons exposed to oxygen-glucose deprivation (OGD), the effects of GAS (500 μmol/L), nutlin-3 (a P53 agonist; 10 μmol/L) or their combination were examined on ferroptosis-related protein expressions, intracellular ROS, lipid peroxidation, MDA, GSH, cell viability, mitochondrial membrane potential, and Fe²⁺ levels. Results In the rat models of HH, GAS treatment significantly inhibited P53 expression, upregulated SLC7A11 and GPX4 proteins, markedly reduced Fe²⁺, ROS, and MDA levels, and increased GSH content in the cerebral cortex. In cultured HT22 neurons, GAS treatment effectively alleviated OGD-induced cell ferroptosis as shown by decreased P53 expression, increased SLC7A11 and GPX4 expressions, and lowered levels of intracellular ROS generation, lipid peroxidation, and Fe²⁺ accumulation, along with obvious restoration of GSH levels, cell viability, and mitochondrial membrane potential. The protective effects of GAS was markedly attenuated by activation of the P53 pathway using nutlin-3. Conclusion GAS produces neuroprotective effects against HH-induced brain injury in rats by inhibiting neuronal ferroptosis via regulating the P53/SLC7A11/GPX4 signaling pathway.

Key words: gastrodin, hypobaric hypoxia-induced brain injury, oxygen-glucose deprivation, ferroptosis, GPX4/SLC7A11/FTH1

Jiping YANG, Yongzeng GU, Zhenyi FU, Yuexin DONG, Yihao DENG, Zheng LI, Liangli LIU, Tao GUO. Gastrodin alleviates hypobaric hypoxia-induced brain injury in rats by reducing neuronal ferroptosis via the P53/SLC7A11/GPX4 signaling axis[J]. Journal of Southern Medical University, 2026, 46(7): 1498-1508.

Figures/Tables 6

Fig.1 Effect of gastrodin (GAS) on expressions of ferroptosis-related proteins P53, SLC7A11, and GPX4 in rat models exposed to hypobaric hypoxia (HH). A: Western blotting for detecting protein expressions of P53, SLC7A11, and GPX4 in cerebral cortical tissues of the rats. B-D: Quantitative analysis of the relative expression levels of the proteins. ***P<0.001 vs normal control (Nor) group; #P<0.05, ##P<0.01, ###P<0.001 vs HH group (n=3).

Fig.2 Effect of GAS on ferroptosis markers P53 and 4-HNE in the cortical neurons of rat models of HH. A, C: Expressions of P53 and 4-HNE (red) in the neurons (NeuN, green) detected by double immunofluorescence staining (Scale bar=10 μm). B, D: Quantitative analysis of fluorescence intensity of P53 and 4-HNE. **P<0.001, ***P<0.001 vs Nor group; #P<0.05, ##P<0.01 vs HH group (n=3).

Fig.3 Effects of GAS on oxidative stress and ferrous ion levels in the cortical tissues of rat models of HH. A, B: ROS levels detected by DHE fluorescence staining and quantitative analysis of fluorescence intensity (Scale bar=10 μm). C: Relative MDA content. D: Relative GSH content. E: Relative ferrous ion concentration. ***P<0.001 vs Nor group; #P<0.05, ##P<0.01, ###P<0.001 vs HH group (n=3).

Fig.4 GAS regulates expressions of SLC7A11 and GPX4 via the P53 pathway in cultured HT22 neurons exposed to oxygen-glucose deprivation (OGD). A, B: Expression of P53 (red) in the neurons (NeuN, green) detected by immunofluorescence staining and quantitative analysis of fluorescence intensity (Scale bar=50 μm). C: Western blotting for detecting P53, SLC7A11, and GPX4 proteins. D-F: Quantitative analysis of relative expression levels of P53, SLC7A11, and GPX4. ***P<0.001 vs Control group; #P<0.05, ##P<0.01, ###P<0.001 vs OGD group; &P<0.05, &&P<0.01, &&&P<0.001 vs OGD+GAS group (n=3).

Fig.5 GAS regulates oxidative stress and lipid peroxidation via the P53 pathway in the OGD cell model. A, C: ROS levels detected by DCFH-DA fluorescence staining and quantitative analysis of fluorescence intensity (Scale bar=50 μm). B, D: Lipid peroxidation levels detected by BODIPY-C11 fluorescence staining and quantitative analysis of the oxidized/reduced ratio (Scale bar=50 μm). E: Relative MDA content. F: Relative GSH content. ***P<0.001 vs Control group; #P<0.05, ##P<0.01, ###P<0.001 vs OGD group; &&P<0.01, &&&P<0.001 vs OGD+GAS group (n=3).

Fig.6 GAS regulates cell survival, mitochondrial membrane potential, and Fe2+ levels via the P53 pathway in the OGD cell model. A: Quantitative analysis of cell viability. B, C: Mitochondrial membrane potential detected by JC-1 staining and quantitative analysis of the aggregates/monomers ratio. D, E: Fe2+ levels detected by FerroOrange fluorescent probe staining and quantitative analysis of fluorescence intensity. Scale bar=50 μm. ***P<0.001 vs Control group; #P<0.05, ###P<0.001 vs OGD group; &&P<0.01, &&&P<0.001 vs OGD+GAS group (n=3).

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