Preparation and characterization of antitumor polysaccharide from Acanthopanax senticosus: A combination of ultrasonic-assisted enzymatic extraction and macroporous resin purification

Recently, rapid and effective extraction and purification methods for polysaccharides have attracted considerable interest as alternatives to traditional methods. The study applied ultrasonic-assisted enzymatic extraction (UAEE) along with a macroporous resin purification (MRP) step to successfully enrich purified Acanthopanax senticosus polysaccharide (PASP) in the extract of Acanthopanax senticosus , examining its structural characteristics as well as biological activity. According to the results, the yield of Acanthopanax senticosus polysaccharide (ASP) was 4.863 ± 0.122 % under optimal extraction conditions, which included a cellulase dosage of 5 %, an ultrasonic temperature of 50℃, and a water-material ratio of 35 g/g, closely matching the predicted value (4.976 %). Under ideal purification conditions, which include a loading flow rate of 2 mL/min, a loading volume of 30 mL, and an elution flow rate of 3 mL/min, the comprehensive membership value calculated by fuzzy membership function (FMF) was 2.70 ± 0.21, approaching the predicted value (2.68), and the purity of PASP reached 87.66 ± 0.84 %. The molecular weight of PASP is 1.146 × 10 4 Da, with its primary components being mannose, rhamnose, galacturonic acid, glucose, galactose, xylose and arabinose in the molar proportions of 9.75:8.40:4.25:34.53:25.84:5.44:11.80. The results of fourier-transform infrared spectroscopy (FTIR) indicate that α and β glycosidic bonds may coexist in PASP. Both x-ray diffraction (XRD) and scanning electron microscopy (SEM) results demonstrate that PASP is an amorphous structure. Additionally, PASP can significantly affect the viability of CT-26 cells by inducing apoptosis and inhibiting cell migration in vitro . Animal experimental results showed that PASP could effectively inhibit the proliferation of CT-26 solid tumors and increase the content of cytokines TNF-α and IL-2. HE staining showed that PASP promoted the apoptosis of tumor cells, and western blotting showed that PASP regulated the expression of Bax, Caspase 3 and Bcl-2. These findings will offer a theoretical foundation and innovative concepts for advancing and utilizing PASP in biomedicine. • Ultrasound-assisted enzymatic extraction of Acanthopanax senticosus polysaccharide was first performed. • Optimization of purification conditions of Acanthopanax senticosus polysaccharide with macroporous resin. • The structure characterizations of purified Acanthopanax senticosus polysaccharide was analyzed. • The antitumor activity of purified Acanthopanax senticosus polysaccharide was investigated in vitro and in vivo.