This study aimed to isolate, purify, and characterize Polygonatum polysaccharides (PSPs) and evaluate their hepatoprotective effects against CCl4-induced liver damage in HepG2 cells. The PSPs were obtained from an aqueous extract of Polygonatum using DEAE cellulose column chromatography, CL-6B agarose gel chromatography, and Sephadex G100 chromatography. Three homogeneous fractions, PSP-N-b-1, PSP-N-b-2, and PSP-N-c-1, were identified, each composed of six monosaccharides: Man (mannose), GlcA (glucuronic acid), Rha (rhamnose), GalA (galacturonic acid), Glc (glucose), and Ara (arabinose). The molecular weights of these fractions were 6.3 KDa, 5.78 KDa, and 3.45 KDa, respectively. The hepatoprotective effects of these fractions were assessed in vitro by evaluating their impact on CCl4-induced liver damage in HepG2 cells. The results showed that the fractions significantly reduced the degree of cell shrinkage, restored cell structures, and increased cell survival rates. Additionally, they reduced the activities of ALT and AST enzymes, indicating reduced liver damage. The fractions also decreased oxidative stress markers (MDA) and inflammatory cytokines (TNF-α, IL-1β, and IL-6), suggesting their anti-inflammatory and antioxidant properties. The study concluded that Polygonatum polysaccharides, particularly PSP-N-c-1, exhibit promising hepatoprotective effects through anti-inflammatory and antioxidant mechanisms. These findings suggest that Polygonatum polysaccharides could be a valuable component in the development of natural dietary supplements and drugs for preventing and treating liver injuries.This study aimed to isolate, purify, and characterize Polygonatum polysaccharides (PSPs) and evaluate their hepatoprotective effects against CCl4-induced liver damage in HepG2 cells. The PSPs were obtained from an aqueous extract of Polygonatum using DEAE cellulose column chromatography, CL-6B agarose gel chromatography, and Sephadex G100 chromatography. Three homogeneous fractions, PSP-N-b-1, PSP-N-b-2, and PSP-N-c-1, were identified, each composed of six monosaccharides: Man (mannose), GlcA (glucuronic acid), Rha (rhamnose), GalA (galacturonic acid), Glc (glucose), and Ara (arabinose). The molecular weights of these fractions were 6.3 KDa, 5.78 KDa, and 3.45 KDa, respectively. The hepatoprotective effects of these fractions were assessed in vitro by evaluating their impact on CCl4-induced liver damage in HepG2 cells. The results showed that the fractions significantly reduced the degree of cell shrinkage, restored cell structures, and increased cell survival rates. Additionally, they reduced the activities of ALT and AST enzymes, indicating reduced liver damage. The fractions also decreased oxidative stress markers (MDA) and inflammatory cytokines (TNF-α, IL-1β, and IL-6), suggesting their anti-inflammatory and antioxidant properties. The study concluded that Polygonatum polysaccharides, particularly PSP-N-c-1, exhibit promising hepatoprotective effects through anti-inflammatory and antioxidant mechanisms. These findings suggest that Polygonatum polysaccharides could be a valuable component in the development of natural dietary supplements and drugs for preventing and treating liver injuries.