In this study, we explored an efficient method for synthesizing sulfated Pleurotus ostreatus polysaccharides (SPOP) with a higher degree of substitution. The structure conformation was characterized and confirmed by the Fourier transform infrared (FT-IR) spectroscopy analysis. A three-factor-three-level Box-Behnken design (BBD) was successfully applied to optimize the SPOP synthesis process. Under the optimal conditions, including a ratio of pyridine to chlorosulfonic acid of 5.10:1, a reaction temperature of 59.92°C, and a reaction time of 3.05 h, the predicted maximum degree of sulfate substitution (DS) reached 0.423 ± 0.006. Furthermore, we evaluated the in vitro radical scavenging abilities and in vivo anti-acute liver injury (anti-ALI) effects of SPOP. The results showed that SPOP demonstrated significantly superior hepatoprotective effects against CCl4-induced liver damage, specifically by enhancing antioxidant activities both in vitro and in vivo, as well as improving hepatic functions. Our findings suggested that SPOP exhibited significant potential as a natural therapeutic agent for ALI and its associated complications, and contributed to the potential antioxidant capacities.
Keywords: hepatoprotection; in vitro antioxidant; optimization; sulfated Pleurotus ostreatus polysaccharides.
© 2025 Wiley‐VHCA AG, Zurich, Switzerland.