Resveratrol mediates mitochondrial function through the sirtuin 3 pathway to improve abnormal metabolic remodeling in atrial fibrillation

Abstract

This study investigated the impact of resveratrol on abnormal metabolic remodeling in atrial fibrillation (AF) and explored potential molecular mechanisms. An AF cell model was established by high-frequency electrical stimulation of HL-1 atrial muscle cells. Resveratrol concentrations were optimized using CCK-8 and flow cytometry. AF-induced increases in ROS and mitochondrial calcium, along with decreased adenosine triphosphate (ATP) and mitochondrial membrane potential, were observed. Resveratrol mitigated these changes and maintained normal mitochondrial morphology. Moreover, resveratrol acted through the SIRT3-dependent pathway, as evidenced by its ability to suppress AF-induced acetylation of key metabolic enzymes. SIRT3 overexpression controls acetylation modifications, suggesting its regulatory role. In conclusion, resveratrol's SIRT3-dependent pathway intervenes in AF-induced mitochondrial dysfunction, presenting a potential therapeutic avenue for AF-related metabolic disorders. This study sheds light on the role of resveratrol in mitigating AF-induced mitochondrial remodeling and highlights its potential as a novel treatment for AF.

Figure 1.

Effect of resveratrol on HL-1 cell viability and apoptosis. A) The CCK-8 method was used to evaluate the effects of the control group and AF group on HL-1 cell viability. B) The CCK-8 method was used to evaluate the effects of resveratrol intervention with different concentration gradients (0, 0.1, 1, 10, 50 μmol/L) on the proliferation of HL-1 cells. C) FCM was used to detect the effect of resveratrol intervention with different concentration gradients (0, 0.1, 1, 10, 50 μmol/L) on the apoptosis of HL-1 cells; *p<0.05, **p<0.01, ***p<0.001; n=3.

Figure 2.

Effects of resveratrol on HL-1 cell viability and apoptosis. A) ATP assay kit evaluated ATP content in HL-1 cells. B) DCFH-DA probe labeled intracellular ROS, and flow cytometry was used to detect the representative image and quantification of fluorescence intensity. C) The MMP level was detected by JC-1 staining, and the red/green fluorescence ratio reflected the change in MMP in HL-1 cells. D) The mitochondrial Ca²⁺ level and fluorescence intensity were quantitatively evaluated by the calcium indicator X-Rhod-1. E) Morphological changes in mitochondria in HL-1 cells were observed by transmission electron microscopy; *p<0.05, **p<0.01, ***p<0.001; n=3.

Figure 3.

Resveratrol regulates the acetylation of metabolic enzymes through a SIRT3-dependent pathway during AF. A) The mRNA expression level of SIRT3 in HL-1 cells was detected by RT-qPCR. B,C) Western blot analysis was used to detect the protein expression level of SIRT3 in HL-1 cells and analyze its gray value. D,G) Western blotting was used to detect the acetylated protein level and gray value analysis of key metabolic enzymes (LCAD, GDH, AceCS2) in HL-1 cells; *p<0.05, **p<0.01, ***p<0.001; n=3.

Figure 4.

Resveratrol mediates mitochondrial function through the sirtuin 3 pathway to improve AF-induced metabolic remodeling. A) The proliferation of HL-1 cells was evaluated by the CCK-8 method. B) FCM was used to detect HL-1 cell apoptosis. C) ATP detection kit to evaluate ATP content in HL-1 cells. D) DCFH-DA probe-labeled intracellular ROS, and flow cytometry was used to detect the representative image and quantification of fluorescence intensity. E) The MMP level was detected by JC-1 staining, and the red/green fluorescence ratio reflected the change in MMP in HL-1 cells. F) The mitochondrial Ca²⁺ level and fluorescence intensity were quantitatively evaluated by the calcium indicator X-Rhod-1. G) Morphological changes in mitochondria in HL-1 cells were observed by transmission electron microscopy; *p<0.05, **p<0.01, ***p<0.001; n=3.