Ferrostatin-1 Reversed Chronic Intermittent Hypoxia-Induced Ferroptosis in Aortic Endothelial Cells via Reprogramming Mitochondrial Function

Jia Chen; Xiaoyu Deng; Ting Lin; Jiefeng Huang; Yisong Yang; Ningfang Lian

doi:10.2147/NSS.S442186

Ferrostatin-1 Reversed Chronic Intermittent Hypoxia-Induced Ferroptosis in Aortic Endothelial Cells via Reprogramming Mitochondrial Function

Nat Sci Sleep. 2024 Apr 23:16:401-411. doi: 10.2147/NSS.S442186. eCollection 2024.

Authors

Jia Chen^#^{1

2

3

4}, Xiaoyu Deng^#^{1

2

3

4}, Ting Lin^{1

2

3

4}, Jiefeng Huang^{1

2

3

4}, Yisong Yang^{1

2

3

4}, Ningfang Lian^{1

2

3

4}

Affiliations

¹ Department of Respiratory and Critical Care Medicine, the First Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian, People's Republic of China.
² Fujian Provincial Sleep-Disordered Breathing Clinic Center, Fuzhou, Fujian, People's Republic of China.
³ Institute of Respiratory Disease, Fujian Medical University, Fuzhou, Fujian, People's Republic of China.
⁴ Department of Respiratory and Critical Care Medicine, National Regional Medical Center, Binhai Campus of the First Affiliated Hospital, Fujian Medical University, Fuzhou, Fujian, People's Republic of China.

^# Contributed equally.

Abstract

Purpose: Chronic intermittent hypoxia (CIH) related arterial endothelium injury is a common cause of cardiovascular system injury. However, the mechanism still needs to be clarified. In this study, we aimed to clarify the role and mechanism of ferrostatin-1 (Fer-1) in CIH-related rat arterial endothelial cells (ROAEC) ferroptosis.

Methods: ROAEC was divided into control group, CIH group, and CIH+ Fer-1 group. Cell viability was detected by cell counting kit 8 kits (CCK8). The apoptotic rate, reactive oxygen species (ROS) levels, Fe2+ levels, and lipid ROS levels were detected by flow cytometry. Malondialdehyde (MDA) levels and nicotinamide adenine dinucleotide (NAD+)/NADH ratio were detected via Elisa kits. The mRNA and protein levels of cystine/glutamate antiporter solute carrier family 7 member 11 (SLC7A11) and glutathione peroxidase 4 (GPX4) were detected by qRT-PCR and Western blot. Mitochondrial structure and function were observed by transmission electron microscope (TEM) and mitochondrial membrane potential (MMP). Central carbon metabolism was measured to compare metabolites among each group.

Results: After the CIH exposure, ROAEC cell viability decreased; The levels of cell apoptosis, ROS, Fe2+, MDA, and lip ROS increased; The levels of NAD+/NADP ratio decreased; The mRNA and protein levels of GPX4 and SLC7A11 decreased (all p<0.05). Co-cultured with Fer-1 reversed the levels of apoptosis rate, cell viability, ROS, Fe2+, MAD, lipid ROS, NAD+/NADH ratio and the mRNA and protein expression of GPX4 and SLC7A11 (all p<0.05). The TEM results showed that damaged mitochondrial membrane and the matrix spillover in the CIH group. The results of the JC-1 assay showed decreased MMP in the CIH group. Fer-1 treatment ameliorated the mitochondrial injury. The results of central carbon metabolism found that CIH altered the metabolites in the TCA cycle, which were reversed by Fer-1 treatment.

Conclusion: CIH-induced ferroptosis in ROAEC, which were reversed by Fer-1 via reprogramming mitochondrial function.

Keywords: arterial endothelium injury; chronic intermittent hypoxia; ferroptosis; mitochondrial function; obstructive sleep apnea.

Grants and funding

This study was funded by the Natural Science Foundation of Fujian Province (No. 2023J01554), Joint Funds for the Fujian Provincial Finance Project (No: BPB-LNF2021), National Natural Science Foundation of China (No. 82170101).