Green synthesis of metal nanoparticles is a rapidly growing research area in the field of nanotechnology because of their biomedical applications. This study describes the synthesis of silver nanoparticles (AgNPs) using Spilanthes acmella leaf extract and its ameliorative effects against doxorubicin-induced toxicity. The formation of AgNPs was confirmed by a ultraviolet-visible (UV-vis) spectrum that revealed an absorption band at 430 nm. A shift in the absorption bands in Fourier-transform infrared spectroscopy (FT-IR) confirmed the bioactive molecules of S. acmella leaf extract that acted as a reducing and capping agent. The spherical shape of AgNPs was confirmed by scanning electron microscope (SEM) analysis, and the presence of elemental silver was indicated by energy dispersive X-ray spectroscopy (EDS) analysis. X-ray diffraction (XRD) analysis revealed that the crystalline size of the synthesized AgNPs was 6.702 nm. Treatment of Dalton's lymphoma ascites (DLA) mice with 20 mg/kg of doxorubicin (DOX) significantly increased the activities of serum toxicity markers including aspartate amino-transferase (AST), alanine amino-transferase (ALT), and lactate dehydrogenase (LDH). However, compared to DOX alone treatment, the coadministration of DOX and AgNPs reduced AST, ALT, and LDH activities. DOX alone treatment reduced glutathione (GSH) contents and decreased the activities of glutathione-s-transferase (GST) and superoxide dismutase (SOD) in DLA mice. However, the administration of AgNPs to DOX-treated DLA mice increased GSH content and the activities of GST and SOD. Consistently, biosynthesized AgNPs were found to possess significantly higher free-radical scavenging activities when compared to the S. acmella leaf extract, as measured by ABTS, DPPH, and O2 •- assays. The biosynthesized AgNPs also showed significant inhibitory activities against erythrocyte hemolysis and lipid peroxidation in the liver homogenate.