Fig 1: Aloe emodin reduces SH3GLB1‐mediated mitoROS accumulation to inhibit NDP52‐induced mitophagy for RIHD protection. A) TEM analysis of H9C2 cells after X‐ray or aloe emodin treatment, yellow arrow for mitochondria, red arrow for autophagic vesicle or autophagosome. Scale bar, 5 µm, 2 µm, and 500 nm. B) Mitochondrial ROS, mitochondrial morphology, and cardiac cytoskeleton and morphology were detected by mtSOX (DOJinDO, MT14), mitoTracker Green (Beyotime, C1048), and actin‐Tracker Green‐488 (Beyotime, C2201S), respectively. Scale bar, 20 µm. C,D) Co‐immunoprecipitation assay of interaction between LC3 and NDP52. E) Co‐localization between NDP52 and Parkin observing by confocal microscopy. Scale bar, 50 µm. F) Knockdown of SH3GLB1 combined with aloe emodin inhibited the interaction between NDP52 and FIP200, ULK1, or ATG13, detected by Western blotting. G,H) Intracellular concentration of ATP and ADP were analyzed by using ATP assay kit (Beyotime, S0026) and ADP assay kit (Abcam, ab83359). I) Blue native page (Beyotime, P0545S) for Western blotting detection of mitochondrial complexes, including complex I, II, III, IV, and V, using a mixture of five antibodies, after mitochondrial dissociation. J) Partial restoration of mitochondrial oxidative respiratory function under SH3GLB1 deletion combined with aloe emodin, using OCR detection. Quantification data are shown as mean ± SEM. All data were obtained from three independent experiments. p‐Values were calculated by unpaired two‐tailed Student's t‐test or one‐way ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001; ns: not significant.