The impairment of autophagy involves oxidative stress-induced cellular senescence, leading to endothelial dysfunctions and the onset of cardiovascular diseases. As molybdenum disulfide nanoparticles (MoS₂ NPs), representative transition metal dichacogenide materials, have great potential as a multifunctional therapeutic agent against various disorders, the present study aimed to investigate whether MoS₂ NPs prevents hydrogen peroxide (H₂O₂)-induced endothelial senescence by modulating autophagic process. Our results showed that pretreatment with MoS₂ NPs inhibited H₂O₂-induced endothelial senescence and improved endothelial functions. Exposure of H₂O₂ increased p62 level and blocked the fusion of autophagosomes with lysosomes, indicating of impaired autophagic flux in senescent endothelial cells. However, MoS₂ NPs pretreatment efficiently suppressed cellular senescence through triggering autophagy and resisting impaired autophagic flux. Furthermore, the genetic inhibition of autophagy by siRNA against Beclin 1 or ATG-5 directly abrogated the protective action of MoS₂ NPs on endothelial cells against H₂O₂-induced senescence.Thus, these results suggested that MoS₂ NPs rescue endothelial cells from H₂O₂-induced senescence by improving autophagic flux, and provide valuable information for the rational design of MoS₂-based nanomaterials for therapeutic use in senescence-related diseases.