3-n-Butylphthalide Protects SH-SY5Y Cells from Ferroptosis by Inhibiting ACSL4-Mediated Lipid Peroxidation

Area of Science:

• Neuroscience
• Biochemistry
• Cell Biology

Background:

• Parkinson's disease (PD) pathogenesis involves oxidative stress, mitochondrial dysfunction, and iron-dependent cell death.
• Previous studies indicated 3-n-butylphthalide (NBP) alleviates PD behavioral deficits, but mechanisms were unclear.

Purpose of the Study:

• To elucidate the underlying mechanisms of NBP's neuroprotective effects in ferroptosis.
• To investigate NBP's interaction with ACSL4 and its role in ferroptosis.

Main Methods:

• Cell viability assays, reactive oxygen species (ROS) measurement, mitochondrial oxidative stress assessment, and lipid peroxidation analysis.
• Molecular docking, cellular thermal shift assay (CETSA), site-directed mutagenesis, and pulse-chase experiments.
• ACSL4 overexpression studies in ferroptosis-induced SH-SY5Y cells.

Main Results:

• NBP treatment attenuated Erastin/RSL3-induced ferroptosis in SH-SY5Y cells, improving viability and reducing oxidative stress markers.
• NBP was found to interact with ACSL4, enhancing its protein stability via specific residues (PRO-404, ILE-526).
• Overexpression of ACSL4 abolished NBP's protective effects, indicating ACSL4's crucial role.

Conclusions:

• NBP protects against ferroptosis by suppressing ACSL4-mediated lipid peroxidation.
• NBP demonstrates therapeutic potential for neurodegenerative diseases like Parkinson's disease associated with ferroptosis.