Melatonin Inhibiting Neuronal Cells Ferroptosis Through Lipid Metabolic Reprogramming

Area of Science:

• Neuroscience
• Molecular Biology
• Biochemistry

Background:

• Ferroptosis is a regulated form of cell death implicated in neurodegenerative diseases.
• Melatonin is a hormone with known antioxidant and neuroprotective properties.
• The precise molecular mechanisms underlying melatonin's neuroprotective effects, particularly concerning ferroptosis, require further elucidation.

Purpose of the Study:

• To investigate the underlying mechanism by which melatonin protects neurons against ferroptosis.
• To identify key molecular players and pathways involved in melatonin's neuroprotective action.
• To elucidate the role of lipid metabolism and the Atf4/Trib3 axis in melatonin-mediated ferroptosis inhibition.

Main Methods:

• Cell culture (HT22 cells) treated with melatonin, Erastin, and Ferrostatin-1.
• Transcriptomic and metabolomic analyses to identify hub genes and pathways.
• Gene Set Enrichment Analysis (GSEA) for pathway and biological process determination.
• Quantitative real-time PCR (qRT-PCR) and Western Blot (WB) for gene and protein expression validation.
• Cell Counting Kit-8 (CCK-8) assay, ROS analysis, and WB for functional confirmation.

Main Results:

• Melatonin significantly inhibited ferroptosis in HT22 cells, increasing GPX4 activity and decreasing reactive oxygen species (ROS) generation.
• Transcriptomic analysis identified Tribble 3 (Trib3) as a melatonin-related hub gene, upregulated by Erastin.
• Lipidomic analysis indicated that melatonin's regulation of lipid metabolism involves glycerophospholipids.
• Melatonin reduced the protein levels of Atf4 and Trib3, suggesting the Atf4/Trib3 axis is a key target.
• Melatonin treatment increased cell survival and GPX4 activity while reducing ROS content.

Conclusions:

• Melatonin exerts neuroprotection by inhibiting ferroptosis through the modulation of lipid metabolism.
• The Atf4/Trib3 signaling axis plays a critical role in mediating melatonin's protective effects against ferroptosis.
• These findings highlight a novel mechanism for melatonin's therapeutic potential in neurological conditions associated with ferroptosis.