This study isolated and purified two polysaccharides, CYHSP-1 and CYHSP-2, from Corydalis yanhusuo, and investigated their structures and in vitro biological activities. The findings revealed that CYHSP-1 and CYHSP-2 possess average molecular weights of 97.17 and 23.48 kDa, respectively, indicating uniform molecular weight glucans. Structural characterization, utilizing techniques such as Fourier transform infrared spectroscopy (FT-IR) and methylation analysis, identified the presence of sugar units including α-1,4-Glc-(1→, α-1,4,6-Glc-(1→, α-1,6-Glc-(1→, and α-t-Glc-(1→. CYHSP-1 demonstrated superior radical scavenging activity and a more pronounced inhibitory effect on human hepatocellular carcinoma HepG2 cells compared to CYHSP-2. These results indicate the significant potential of these Corydalis yanhusuo-derived polysaccharides as natural antioxidants and anti-tumor agents.