南瓜籽多糖的分离纯化及其体外降血糖活性研究  

Isolation,Purification,and in Vitro Hypoglycemic Activity of Pumpkin Seed Polysaccharides

作　　者：蔡舒 钟诚 黎敏姿 罗钧匀 周爱梅[1] CAI Shu;ZHONG Cheng;LI Minzi;LUO Junyun;ZHOU Aimei(Guangdong Provincial Key Laboratory of Nutraceuticals and Functional Foods,College of Food Science,South China Agricultural University,Guangzhou 510642,China)

机构地区：[1]华南农业大学食品学院,广东省功能食品活性物重点实验室,广东广州510642

出　　处：《食品工业科技》2025年第9期352-362,共11页Science and Technology of Food Industry

基　　金：广州市科技计划项目(202206010129)。

摘　　要：南瓜籽提油后会产生大量废弃饼粕,而多糖是南瓜籽粕主要活性成分之一,为减少资源浪费并充分利用其价值,该研究以提油后的南瓜籽粕为原料,经连续相变萃取装置大批量提取南瓜籽粗多糖后,采用95%乙醇醇沉、Sevage法与酶法相结合除蛋白、DEAE-52和Experdex-75柱进行分离纯化,最后结合体外酶活抑制实验和3T3-L1脂肪细胞实验探究与比较各组分的降血糖活性,并初步探索其降糖机制。研究发现,南瓜籽多糖纯化后得到5种多糖组分(PSP、PSP-1、PSP-1-1、PSP-1-2、PSP-1-3),其中PSP-1-1、PSP-1-2、PSP-1-3的单糖组成及摩尔比有一定差异。体外α-淀粉酶和α-葡萄糖苷酶抑制实验表明,效果最佳的PSP-1-1组分的IC_(50)值分别为0.88 mg/mL和3.92 mg/mL。细胞实验则表明,PSP-1-1显著提高细胞摄取葡萄糖能力,同时极显著提高高密度脂蛋白胆固醇(HDL-C)含量至257%,而甘油三酯(TG)、总胆固醇(TC)、低密度脂蛋白胆固醇(LDL-C)含量分别降低了48.65%、55.91%、50%,说明其能通过改善糖脂代谢来缓解细胞的胰岛素抵抗。并猜测其降血糖机制为PSP-1-1通过促进胰岛素受体底物(IRS-1)/磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(PKB,也称AKT)通路中基因的表达,恢复细胞正常代谢,达到降血糖效果。该研究提高了南瓜籽副产物的综合利用价值,为开发毒副作用低的降糖功能食品提供了理论支撑。After oil extraction,pumpkin seeds generate a large amount of waste meal,and polysaccharides are one of the main active components in pumpkin seed meal.To mitigate resource waste and maximize its value,pumpkin seed meal was used as raw material in this study to extract crude polysaccharides in large quantities by continuous phase transition extraction device.The crude polysaccharides were then subjected to precipitation using 95%ethanol and deproteinization using the Sevage method combined with enzymatic treatment,followed by further separation and purification via DEAE-52 and Experdex-75 columns.Finally,in vitro enzyme activity inhibition assays and 3T3-L1 adipocyte experiments were conducted to investigate and compare the hypoglycemic activity and underlying mechanism of the resulted various components.The study revealed that five polysaccharide fractions(PSP,PSP-1,PSP-1-1,PSP-1-2,PSP-1-3)were obtained after purification of pumpkin seed polysaccharides.The monosaccharide composition and molar ratio of PSP-1-1,PSP-1-2 and PSP-1-3 were different.In vitroα-amylase andα-glucosidase inhibition assays showed PSP-1-1 exhibited the most potent effects with IC_(50) values of 0.88 mg/mL and 3.92 mg/mL,respectively.Cellular experiments demonstrated PSP-1-1 significantly enhanced the ability to uptake glucose by cells and markedly increased the high-density lipoprotein cholesterol(HDL-C)level to 257%,while reduced the levels of triglyceride(TG),total cholesterol(TC),and low-density lipoprotein cholesterol(LDL-C)by 48.65%,55.91%and 50%,respectively,suggesting it could alleviate cellular insulin resistance by improving glucose and lipid metabolism.It was hypothesized that the hypoglycemic mechanism of PSP-1-1 involved promoting the expression of genes in the insulin receptor substrate(IRS-1)/phosphatidylinositol 3-kinase(PI3K)/protein kinase B(PKB,also known as AKT)pathway,restoring normal cellular metabolism,and ultimately achieving hypoglycemic effect.This study enhances the comprehensive utilization value of pumpkin see

关 键 词：南瓜籽多糖 分离纯化 酶活力 3T3-L1 脂肪细胞 降血糖 作用机制 

分 类 号：TS255.1[轻工技术与工程—农产品加工及贮藏工程]