Ferroptosis is a form of programmed cell death that is caused by iron ion-dependent accumulation of lipid peroxides. Ferroptosis has been shown to follow a different cell death pathway from apoptosis and thus is attracting attention as a new target for cancer therapy. It has also been found to be associated with various diseases, such as neurodegenerative diseases, cerebral apoplexy, and hepatitis (NASH).
Ferroptosis is characterized by the accumulation of lipid peroxides. Lipid peroxides are formed from oxidation of polyunsaturated fatty acids (PUFA) in membrane phospholipids, with iron suggested to be involved. Intracellular glutathione peroxidase 4 (GPX4) uses reduced glutathione (GSH), an antioxidant, to reduce lipid peroxides generated by reactive oxygen species (ROS).
However, when lipid peroxides accumulate due to GPX4 disruption or GSH depletion, ferroptosis is triggered.
Erastin is a known inducer of ferroptosis. By inhibiting the cystine transporter (xCT), erastin inhibits the uptake of cystine. Cystine is the raw material for GSH. Therefore, Erastin ultimately decreases the amount of GSH. Decreased GSH then results in lipid peroxide accumulation and induction of ferroptosis.
The following experimental examples show changes in each index because of erastin stimulation.
Using erastin-treated A549 cells, we measured intracellular Fe2+, ROS, lipid peroxide, glutathione, glutamate release into the extracellular space, and cystine uptake. As a result, inhibition of xCT by elastin was observed and the release of glutamate and uptake of cystine were decreased. Furthermore, elastin treatment decreased intracellular glutathione while it increased intracellular Fe2+, ROS, and lipid peroxides.
|Object||Product Name||CAT. No.|
|①Cystine Uptake||(Under Development)|
|②Released Glutamate||Glutamate Assay Kit-WST||G269-10|
|③Intracellular Glutathione||GSSG/GSH Quantification Kit||G257-10|
|④Intracellular Fe2+||FerroOrange||F374-10; F374-12|
|⑤Intracellular ROS||ROS Assay Kit -Highly Sensitive DCFH-DA-||R252-10|
|⑥Intracellular Lipid Peroxide||Liperfluo||L248-10|
Lipid Peroxide and Iron (Fe2+) Detection Reagents
|Product Name||CAT. No.||Target||Localization||Detection (Fluorescence: Ex/Em)||Instrument||Sample|
|Liperfluo||L248-10||Lipid Peroxidation||Intracellular||Fluorescence (524 nm/535 nm)||Fluorescence Microscope, FCM||Live Cell|
|MitoPeDPP||M466-10||Lipid Peroxidation||Mitochondria||Fluorescence (452 nm/470 nm)||Fluorescence Microscope, FCM||Live Cell|
|Mito-FerroGreen||M489-10||Ferrous Ion (Fe2+)||Mitochondria||Fluorescence (505 nm/580 nm)||Fluorescence Microscope, Microplate Reader||Live Cell|
|FerroOrange||F374-10; F374-12||Ferrous Ion (Fe2+)||Intracellular||Fluorescence (543 nm/580 nm)||Fluorescence Microscope||Live Cell|
Oxidative Stress- and Metabolism-Related Reagents and Kits
|Name||CAT. No.||Target||Localization||Detection (Fluorescence: Ex/Em)||Instrument||Sample|
|ROS Assay Kit -Highly Sensitive DCFH-DA-||R252-10||ROS (Reactive oxygen species)||Intracellular||Fluorescence
(505 nm/525 nm)
|Fluorescence Microscope, FCM, Microplate Reader||Live Cell|
|GSSG/GSH Quantification Kit||G257-10||Glutathione (oxidized/reduced)||Intracellular||Colorimetric:412 nm||Microplate Reader||Cell, Tissue, Blood Plasma, Red Blood Cell|
|Glutamine Assay Kit-WST||G268-10||Glutamine||Intracellular/ Extracellular||Colorimetric:450 nm||Microplate Reader||Cell, Cell Culture|
|Glutamate Assay Kit-WST||G269-10||Glutamine||Intracellular/ Extracellular||Colorimetric:450 nm||Microplate Reader||Cell, Cell Culture|