Fig. 1

Hyperuricemia induces increased mitochondrial fission and dysfunction, elevated oxidative stress, and decreased ATP production in AC16 cardiomyocytes and increased HMGCS2 expression in cardiac tissues and cardiomyocytes. A, B Western blot analysis of FIS1, MFN1, MFN2 (A) protein levels in AC16 cells after uric acid treatment (0, 100, 200, 400 mg/L, 24 h). Statistical analysis (B) is shown (n = 5/group). C, D Mitochondrial morphology in AC16 cells treated with uric acid (400 mg/L, 24 h) was assessed using MitoTracker Red. Representative images (C, scale bar: 10 µm) and ImageJ quantification (D) of the number of mitochondria (count), the mean perimeter (perimeter), the mean aspect ratio (aspect ratio), the mean form factor (form factor), the number of mitochondria branches/mitochondria (Branches/mito) and the branches length/mitochondria (Branch length/mito) are shown. E, F Mitochondrial membrane potential was analyzed using JC-1 staining. Representative fluorescence images of JC-1 aggregates (red) and monomers (green) are presented (E, scale bar: 25 µm), along with quantification of the JC-1 aggregate/monomer ratio (F, n = 5/group). G, H Oxidative stress was detected in AC16 cells using DHE staining after uric acid treatment (400 mg/L, 24 h). Representative fluorescence images (G) and quantification (H, n = 5/group) are provided. I-K Measurements in AC16 cells include relative MDA content (I), SOD activity (J), and ATP levels (K) (n = 5/group). L, M. Volcano plot (L) and heatmap (M) of differentially expressed genes in myocardial tissue from hyperuricemic mice versus controls (n = 4 mice/group). N, O Representative western blot image (N) and corresponding statistical analysis (n = 3 mice/group) (O) of HMGCS2 protein levels in the myocardial tissue of hyperuricemic model mice. P-Q. Western blot analysis of HMGCS2 (P) protein levels in AC16 cells after uric acid treatment (0, 100, 200, 400 mg/L, 24 h). Statistical analysis (Q) is shown (n = 5/group)