吴茱萸多糖的分离纯化、结构表征及体外降脂活性

孙永进; 曾惠梅; 黄静; 赵云龙; 蔡锦源; 孙松

doi:10.13386/j.issn1002-0306.2024070437

吴茱萸多糖的分离纯化、结构表征及体外降脂活性

In vitro Lipid-lowering Activity, Separation and Purification, Structural Characterization of Polysaccharides Fractionated from Evodia Rutaecarpa

摘要

摘要: 为探究吴茱萸多糖的结构特征及其体外降脂的潜力，通过微波辅助提取技术，在吴茱萸中提取得到吴茱萸粗多糖（WPER），利用DEAE-52纤维素离子柱和Sepharose G-100凝胶柱对WPER进行分离纯化，获得了三种多糖组分：WPER-1b、WPER-2a和WPER-3a，最后对三种多糖组分开展了理化特性测定、结构表征以及体外降脂活性研究。理化性质表明三个组分多糖主要由甘露糖（Man）、半乳糖（Gal）和阿拉伯糖（Ara）组成，WPER-2a的总糖含量和糖醛酸含量最高，分别为66.44%±0.64%、32.41%±0.99%。红外表征表明分离纯化不会改变多糖官能团，刚果红实验表明WPER-2a和WPER-3a具有三螺旋结构，三种多糖均以非定型的结晶态存在，热重实验和Zeta电位表明WPER-2a稳定性高于其他两个组分。扫描电镜（SEM）结果表明WPER-1b和WPER-3a为块状和片状，而WPER-2a表现为粗糙、碎末状。体外降脂活性的研究结果表明，WPER-3a浓度10 mg/mL时，对油脂的吸附量最高为22.96 mg/g。WPER-1b、WPER-2a和WPER-3a浓度为10 mg/mL时对胰脂肪酶的抑制率分别为65%、62.33%和64.31%，WPER-2a（10 mg/mL）对胆酸和牛磺胆酸具有较好的吸附作用，分别为75.89%、72.29%，分离纯化后的吴茱萸多糖均具有一定的体外降脂活性，WPER-2a组分活性较好。研究结果为提高吴茱萸多糖的开发利用水平提供参考依据，为吴茱萸多糖在降脂方面的应用提供理论基础和思路借鉴。

Abstract: To explore the structural characteristics of Evodia rutaecarpa polysaccharide and its lipid-lowering potential in vitro, Evodia rutaecarpa crude polysaccharide (WPER) was extracted by microwave-assisted extraction technique. Three fractions of WPER, WPER-1b, WPER-2a and WPER-3a, were isolated and purified by DEAE-52 cellulose ion column and Sepharose G-100 gel column. The physicochemical properties, structure and lipid-lowering activity of the three fractions were characterized. Physicochemical properties showed that the three fractions were mainly composed of Man, Gal and Ara. The total sugar content and uronic acid content of WPER-2a were the highest, which were 66.44%±0.64% and 32.41%±0.99%, respectively. Congo red assay showed that WPER-3a had a triple helix structure. XRD analysis showed that all the three polysaccharides existed in an amorphous crystalline state. Tga and Zeta potential showed that WPER-2a was more stable than the other two polysaccharides. SEM showed that WPER-1b and WPER-3a were bulk and lamellar, while WPER-2a showed a rough, porous, fragmented shape. The results of in vitro lipid-lowering activity study showed that the highest lipid adsorption capacity of WPER-3a was 22.96 mg/g when the concentration of WPER-3a was 10 mg/mL. When the concentration of WPER-1b, WPER-2a and WPER-3a was 10 mg/mL, the inhibition rates of pancreatic lipase were 65%, 62.33% and 64.31%, respectively. WPER-2a (10 mg/mL) had a better adsorption effect on cholic acid and taurocholate, which were 75.89% and 72.29%, respectively. All the isolated and purified polysaccharides of Evodia officinalis had certain lipid-lowering activity in vitro, and the WPER-2a fraction had the better activity. The results provide reference for improving the development and utilization level of Evodia officinalis polysaccharide, and provide theoretical basis and reference for the application of Evodia officinalis polysaccharide in lipid lowering.