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SOX9 depletion attenuates retinal ganglion cell ferroptosis through blocking ERK/p38 signaling
Wei Wei1, Yufeng Zhang1, Langen Li1
1Ophthalmology Department, Inner Mongolia Autonomous Region People's Hospital, Hohhot 010017, Inner Mongolia Autonomous Region, China.
Background:
Retinal ischemia-refusion (I/R) is a leading cause of irreversible blindness worldwide. This study aims to explore the regulatory role of SOX9 in retinal I/R injury, and attempts to elucidate its potential regulatory mechanism.
Methods:
Retinal I/R injury model was established in vivo, and the histological changes was examined by hematoxylin and eosin (H&E) staining and immunofluorescent assay was performed to examine SOX9 expression. Oxygenation-glucose deprivation/reoxygenation (OGD/R)-induced retinal ischemia/reperfusion (I/R) injury in 661 W cells was constructed as an in vitro cellular model of glaucoma. The production of cytokines, lactate dehydrogenase (LDH) and the antioxidant enzymes were assessed by their commercial kits. Cellular reactive oxygen species (ROS) and lipid ROS was detected using DCFH-DA and C11-BODIPY 581/591 staining, respectively. Lipid peroxidation and Fe2+ level were detected to assess the ferroptosis level. Protein expression was examined by western blot. LM22B-10, the agonist of ERK signaling, was used to pretreat 661 W cells for mechanism investigation.
Results:
SOX9 was aberrantly upregulated following retinal I/R injury both in vivo and in vitro. SOX9 knockdown exerted a protective role against OGD/R-triggered oxidative stress, inflammatory response and ferroptosis in 661 W cells. Further, ERK/p38 signaling was activated in 661 W cells following OGD/R induction, which was repressed by SOX9 knockdown, and the ERK signaling agonist partially counteracted the protective role of SOX9 knockdown against oxidative stress, inflammatory response and ferroptosis in OGD/R-induced 661 W cells.
Conclusion:
Collectively, inhibiting SOX9 to block oxidative stress, inflammation and ferroptosis by inactivating ERK/p38 signaling might be effective to prevent retinal I/R injury, thereby alleviating glaucoma.
Insights
Inhibiting SOX9 protects against retinal ischemia-reperfusion injury by reducing oxidative stress, inflammation, and ferroptosis via the ERK/p38 pathway, offering a potential glaucoma treatment.
Area of Science:
- Ophthalmology
- Molecular Biology
- Cell Biology
Background:
- Retinal ischemia-reperfusion (I/R) injury is a significant cause of irreversible blindness globally.
- Understanding the molecular mechanisms underlying retinal I/R injury is crucial for developing effective treatments.
Purpose of the Study:
- To investigate the regulatory role of SOX9 in retinal I/R injury.
- To elucidate the potential mechanism by which SOX9 influences retinal I/R injury.
Main Methods:
- Established in vivo retinal I/R injury models and in vitro oxygen-glucose deprivation/reoxygenation (OGD/R) models using 661W cells.
- Assessed histological changes, SOX9 expression, cytokine production, lactate dehydrogenase (LDH), oxidative stress markers (ROS, lipid ROS), and ferroptosis markers.
- Utilized western blot for protein expression analysis and an ERK signaling agonist (LM22B-10) to explore mechanistic pathways.
Main Results:
- SOX9 was found to be upregulated in retinal I/R injury both in vivo and in vitro.
- SOX9 knockdown demonstrated a protective effect against OGD/R-induced oxidative stress, inflammation, and ferroptosis in 661W cells.
- ERK/p38 signaling activation was observed post-OGD/R, repressed by SOX9 knockdown; ERK agonist partially reversed the protective effects of SOX9 knockdown.
Conclusions:
- Inhibiting SOX9 can mitigate retinal I/R injury by blocking oxidative stress, inflammation, and ferroptosis.
- The inactivation of ERK/p38 signaling by SOX9 inhibition is a key mechanism in protecting against retinal I/R injury.
- Targeting SOX9 represents a potential therapeutic strategy for preventing retinal I/R injury and alleviating glaucoma.
