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1- Free GSH=DTNB+Sample+Substrate 2- Total GSH=DTNB+Sample+Substrate+ NADPH+glutathione reductase 3- GSSG=Total GSH-Free GSH Another method to measure GSSG separately, is to remove the free GSH from DTNB reaction process by modification of free GSH with N-ethylmaleimide (NEM) or vinylpyridine prior to conducting assay

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In the tMCAO rat model, CHY improved neurological function and reduced infarct size by upregulating SLC7A11 and GPX4, and downregulating ferroptosis markers TfR1, PTGS2, and ACSL4, thereby lowering total iron, lipid peroxidation, and MDA levels, and mitigating oxidative stress