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Ketone ester and β-OHB mitigate lipid peroxidation and iron accumulation. ( A ) Representative fluorescence images of C11-BODIPY staining in HASMCs from various treatment groups: Control, β-OHB, RSL3, and RSL3 + β-OHB treatment groups, showing both reduced (red) and oxidized (green) lipid signals. Scale bar = 100 µm. ( B ) Quantitative analysis of the C11-BODIPY fluorescence ratio (Oxidized/Total), indicating the degree of lipid peroxidation. ( C ) Representative fluorescence images of <t>FerroOrange</t> staining in HASMCs used to detect intracellular labile iron (Fe 2+ ) in Control, β-OHB, Fer-1, RSL3, RSL3 + β-OHB, and RSL3 + Fer-1 groups. Scale bar = 100 µm. ( D ) Quantification of FerroOrange Mean Fluorescence Intensity. ( E ) Relative concentration of malondialdehyde (MDA) measured via TBARS assay across the indicated experimental groups in HASMCs. ( F ) Representative images of Prussian blue staining in aortic tissue sections from Control, BAPN, and BAPN + KE mice cohortsblue arrows indicating in vivo iron deposition. Scale bars = 100 µm and 20 µm. ( G ) Quantification of Prussian blue spot density (spots/mm 2 ). Data are presented as mean ± SEM ( n = 3–6 per group). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. β-OHB: 5 mM; Fer-1 (Ferrostatin-1): 2 µM; RSL3: 200 nM (2 h).
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Ketone ester and β-OHB mitigate lipid peroxidation and iron accumulation. ( A ) Representative fluorescence images of C11-BODIPY staining in HASMCs from various treatment groups: Control, β-OHB, RSL3, and RSL3 + β-OHB treatment groups, showing both reduced (red) and oxidized (green) lipid signals. Scale bar = 100 µm. ( B ) Quantitative analysis of the C11-BODIPY fluorescence ratio (Oxidized/Total), indicating the degree of lipid peroxidation. ( C ) Representative fluorescence images of <t>FerroOrange</t> staining in HASMCs used to detect intracellular labile iron (Fe 2+ ) in Control, β-OHB, Fer-1, RSL3, RSL3 + β-OHB, and RSL3 + Fer-1 groups. Scale bar = 100 µm. ( D ) Quantification of FerroOrange Mean Fluorescence Intensity. ( E ) Relative concentration of malondialdehyde (MDA) measured via TBARS assay across the indicated experimental groups in HASMCs. ( F ) Representative images of Prussian blue staining in aortic tissue sections from Control, BAPN, and BAPN + KE mice cohortsblue arrows indicating in vivo iron deposition. Scale bars = 100 µm and 20 µm. ( G ) Quantification of Prussian blue spot density (spots/mm 2 ). Data are presented as mean ± SEM ( n = 3–6 per group). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. β-OHB: 5 mM; Fer-1 (Ferrostatin-1): 2 µM; RSL3: 200 nM (2 h).
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Ketone ester and β-OHB mitigate lipid peroxidation and iron accumulation. ( A ) Representative fluorescence images of C11-BODIPY staining in HASMCs from various treatment groups: Control, β-OHB, RSL3, and RSL3 + β-OHB treatment groups, showing both reduced (red) and oxidized (green) lipid signals. Scale bar = 100 µm. ( B ) Quantitative analysis of the C11-BODIPY fluorescence ratio (Oxidized/Total), indicating the degree of lipid peroxidation. ( C ) Representative fluorescence images of FerroOrange staining in HASMCs used to detect intracellular labile iron (Fe 2+ ) in Control, β-OHB, Fer-1, RSL3, RSL3 + β-OHB, and RSL3 + Fer-1 groups. Scale bar = 100 µm. ( D ) Quantification of FerroOrange Mean Fluorescence Intensity. ( E ) Relative concentration of malondialdehyde (MDA) measured via TBARS assay across the indicated experimental groups in HASMCs. ( F ) Representative images of Prussian blue staining in aortic tissue sections from Control, BAPN, and BAPN + KE mice cohortsblue arrows indicating in vivo iron deposition. Scale bars = 100 µm and 20 µm. ( G ) Quantification of Prussian blue spot density (spots/mm 2 ). Data are presented as mean ± SEM ( n = 3–6 per group). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. β-OHB: 5 mM; Fer-1 (Ferrostatin-1): 2 µM; RSL3: 200 nM (2 h).

Journal: Cells

Article Title: Ketone Ester Attenuates Thoracic Aortic Aneurysm and Dissection by Suppressing Ferroptosis

doi: 10.3390/cells15090829

Figure Lengend Snippet: Ketone ester and β-OHB mitigate lipid peroxidation and iron accumulation. ( A ) Representative fluorescence images of C11-BODIPY staining in HASMCs from various treatment groups: Control, β-OHB, RSL3, and RSL3 + β-OHB treatment groups, showing both reduced (red) and oxidized (green) lipid signals. Scale bar = 100 µm. ( B ) Quantitative analysis of the C11-BODIPY fluorescence ratio (Oxidized/Total), indicating the degree of lipid peroxidation. ( C ) Representative fluorescence images of FerroOrange staining in HASMCs used to detect intracellular labile iron (Fe 2+ ) in Control, β-OHB, Fer-1, RSL3, RSL3 + β-OHB, and RSL3 + Fer-1 groups. Scale bar = 100 µm. ( D ) Quantification of FerroOrange Mean Fluorescence Intensity. ( E ) Relative concentration of malondialdehyde (MDA) measured via TBARS assay across the indicated experimental groups in HASMCs. ( F ) Representative images of Prussian blue staining in aortic tissue sections from Control, BAPN, and BAPN + KE mice cohortsblue arrows indicating in vivo iron deposition. Scale bars = 100 µm and 20 µm. ( G ) Quantification of Prussian blue spot density (spots/mm 2 ). Data are presented as mean ± SEM ( n = 3–6 per group). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. β-OHB: 5 mM; Fer-1 (Ferrostatin-1): 2 µM; RSL3: 200 nM (2 h).

Article Snippet: Intracellular labile ferrous iron (Fe 2+ ) was detected using the live-cell fluorescent probe FerroOrange (Dojindo, Rockville, MD, USA, Cat. No. F374).

Techniques: Fluorescence, Staining, Control, Concentration Assay, TBARS Assay, In Vivo