Xanthohumol Protects the Rat Myocardium against Ischemia/Reperfusion Injury-Induced Ferroptosis

Figure 3

XN prevents ferroptosis-induced myocardial cell death. Treatment with (a) Fe-SP (2 μM) or (b) RSL3 (0.5 μM) for 24 h decreased the viability of neonatal cardiomyocytes examined using the MTT assay, and this effect was inhibited by cotreatment with XN (20 or 50 μM) or a ferroptosis inhibitor (1 μM of Fer-1, or 2 μM of Lip-1) (). (c) Treatment of cardiomyocytes with Fe-SP (2 μM) or RSL3 (0.5 μM) decreased cell number, but did not induce chromatin condensation and necrotic cell death detected by confocal microscopy using Hoechst 33342/PI dye. XN (50 μM) treatment abolished the Fe-SP- or RSL3-induced cell number reduction. H₂O₂ served as a positive control for cell death induction. Scale bars: 100 μm. (d) Scatter plots of necrotic cells (Q1), late apoptotic cells (Q2), early apoptotic cells (Q3), and viable cells (Q4) distinguished by flow cytometric analysis using Annexin V-FITC and propidium iodide staining. (e) Percentage of necrotic cells, late apoptotic cells, and early apoptotic cells and viable were not significant difference between the Fe-SP- or RSL3-treated group and control group. H₂O₂ served as a positive control for cell death induction (). All data represent . , versus the control group; ^#, versus the Fe-SP; ^$, versus the RSL3. Con: control; Fer-1: ferrostatin-1; Lip-1: liproxstatin-1; Hoe: Hoechst 33342; PI: propidium iodide.

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