研究目的
To develop a fluorescence-enabled inhibited autoxidation (FENIX) approach for accurate quantitation of radical-trapping antioxidant activity in phospholipid bilayers, enabling reliable prediction of the anti-ferroptotic potency of redox-active compounds.
研究成果
The FENIX assay provides a superior method for quantifying radical-trapping antioxidant activity in phospholipid bilayers, enabling reliable prediction of anti-ferroptotic potency. The study underscores the importance of H-bonding interactions between antioxidants and phospholipid head groups in determining antioxidant efficacy.
研究不足
The study highlights the limitations of the DPPH assay in predicting anti-ferroptotic potency and the need for assays that accurately reflect the reactivity of antioxidants with lipid peroxyl radicals under physiologically relevant conditions.
1：Experimental Design and Method Selection:
The study involved the development of a fluorescence-enabled inhibited autoxidation (FENIX) approach to quantify radical-trapping antioxidant activity in phospholipid bilayers.
2：Sample Selection and Data Sources:
Mouse embryonic fibroblasts (MEFs) treated with the GPX4 inhibitor RSL3 were used to determine the potency of ferroptosis inhibitors.
3：List of Experimental Equipment and Materials:
Included STY-BODIPY or PBD-BODIPY as signal carriers, egg phosphatidylcholine liposomes, and various radical initiators like AAPH, MeOAMVN, and DTUN.
4：Experimental Procedures and Operational Workflow:
The assay involved monitoring the oxidation of STY-BODIPY or PBD-BODIPY in the presence of antioxidants and calculating inhibition rate constants (kinh) and stoichiometries (n).
5：Data Analysis Methods:
The kinetics of H-atom transfer reactions were analyzed using a pre-dissociation model, and the correlation between kinh values and anti-ferroptotic potency was evaluated.
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