Objective To study the role of silent mating type information regulation 2 homolog-3 (Sirt3) in the process of cardiac senescence induced by advanced glycosylation end-products (AGEs) and to explore the mechanism of total saponins extracted from Dioscorea panthaica (TSDP) in cardiac senescence. Methods After neonatal rat cardiomyocytes were isolated and cultured and identified, the cells were incubated with AGEs. Small interfering RNA (siRNA) was used to knock down the expression of Sirt3 in the cells. Western blotting was used to detect the expression of P16 and P53 and superoxide dismutase-2 (SOD2). SA-β-Gal kit was employed to detect cardiomyocyte senescence, and 2,7-dichlorodi-hydrofluorescein diacetate (DCFH-DA) probe was used to detect intracellular reactive oxygen species (ROS). In the cells with TSDP pre-intervention after AGEs treatment, cardiac senescence was detected by SA-β-Gal kit. GraphPad Prism statistical analysis was performed. One-way ANOVA was conducted for data comparison between two groups. Results Compared with the control cells, the levels of SIRT3 and SOD2 were reduced, and that of P53 was increased, and the proportion of SA-β-Gal positive cells and production of ROS were elevated in the AGEs+negative control (NC) siRNA group. Same changing trends were observed in the AGEs+Sirt3 siRNA group, and the expression level of senescence-related protein P53 was also up-regulated. TSDP pre-intervention decreased the proportion of SA-β-Gal positive cells when compared with the AGEs+NC siRNA group, but the change was decreased after transfection with Sirt3 siRNA. Conclusion AGEs may decrease the expression of SOD2 by down-regulating Sirt3, then aggravate mitochondrial oxidative stress and promote cardiomyocyte aging. TSDP may affect cardiac senescence by regulating the expression of Sirt3.