Inhibition of neuronal ferroptosis protects hemorrhagic brain
Qian Li, Xiaoning Han, Xi Lan, Yufeng Gao, Jieru Wan, Frederick Durham, Tian Cheng, Jie Yang, Zhongyu Wang, Chao Jiang, Mingyao Ying, Raymond C. Koehler, Brent R. Stockwell, Jian Wang
Qian Li, Xiaoning Han, Xi Lan, Yufeng Gao, Jieru Wan, Frederick Durham, Tian Cheng, Jie Yang, Zhongyu Wang, Chao Jiang, Mingyao Ying, Raymond C. Koehler, Brent R. Stockwell, Jian Wang
View: Text | PDF
Research Article Neuroscience

Inhibition of neuronal ferroptosis protects hemorrhagic brain

Abstract

Intracerebral hemorrhage (ICH) causes high mortality and morbidity, but our knowledge of post-ICH neuronal death and related mechanisms is limited. In this study, we first demonstrated that ferroptosis, a newly identified form of cell death, occurs in the collagenase-induced ICH model in mice. We found that administration of ferrostatin-1, a specific inhibitor of ferroptosis, prevented neuronal death and reduced iron deposition induced by hemoglobin in organotypic hippocampal slice cultures (OHSCs). Mice treated with ferrostatin-1 after ICH exhibited marked brain protection and improved neurologic function. Additionally, we found that ferrostatin-1 reduced lipid reactive oxygen species production and attenuated the increased expression level of PTGS2 and its gene product cyclooxygenase-2 ex vivo and in vivo. Moreover, ferrostatin-1 in combination with other inhibitors that target different forms of cell death prevented hemoglobin-induced cell death in OHSCs and human induced pluripotent stem cell–derived neurons better than any inhibitor alone. These results indicate that ferroptosis contributes to neuronal death after ICH, that administration of ferrostatin-1 protects hemorrhagic brain, and that cyclooxygenase-2 could be a biomarker of ferroptosis. The insights gained from this study will advance our knowledge of the post-ICH cell death cascade and be essential for future preclinical studies.

Authors

Qian Li, Xiaoning Han, Xi Lan, Yufeng Gao, Jieru Wan, Frederick Durham, Tian Cheng, Jie Yang, Zhongyu Wang, Chao Jiang, Mingyao Ying, Raymond C. Koehler, Brent R. Stockwell, Jian Wang

×

Figure 2

Fer-1 inhibits ferrous iron–induced neuronal death in OHSCs.

Options: View larger image (or click on image) Download as PowerPoint
Fer-1 inhibits ferrous iron–induced neuronal death in OHSCs. (A–C) Organ...
(A–C) Organotypic hippocampal slice cultures (OHSCs) were treated with 10 μM ferrostatin-1 (Fer-1), 20 μM hemoglobin (Hb) plus vehicle, Hb with Fer-1 (cotreatment), 0.2 mM FeCl2 plus vehicle (Fe2+), FeCl2 with Fer-1 (cotreatment), or vehicle for 16 hours. Then they were stained with propidium iodide (PI) (A), and the percentage of PI+ cells was calculated (B). (C) Culture medium was collected and concentrated for use in the lactate dehydrogenase (LDH) assay. (D and E) OHSCs were treated as described in A–C, fixed, and stained with Fluoro-Jade B (FJB). Quantification of FJB+ cells (D) and representative images from the CA1 region (E) are shown. (F) OHSCs were treated as described in A for 3 days. Slices were fixed and stained with Perls’ stain. Representative images and quantification of iron-positive cells are shown. *P < 0.05, **P < 0.01, ***P < 0.001 versus control; #P < 0.05, ##P < 0.01 versus Hb; †P < 0.05, ††P < 0.01 versus Fe2+. Results are presented as box-and-whisker plots (the middle horizontal line within the box represents the median, boxes extend from the 25th to the 75th percentile, and the whiskers represent 95% confidence intervals). One-way ANOVA followed by Dunn’s multiple comparison post test was used. Results are from at least 3 independent experiments. Scale bars: 1 mm (A), 0.5 mm (E), 50 μm (F).