白茅根多糖理化性质及对HepG2细胞的降糖作用研究  

Physicochemical properties and hypoglycemic effect of polysaccharides from Rhizoma imperatae on HepG2 cells

作　　者：魏菱鸽 李慧[2] 米圣成 路祺[2] 朱明华 WEI Lingge;LI Hui;MI Shengcheng;LU Qi;ZHU Minghua(College of Life Sciences,Northeast Forestry University,Harbin 150040,China;College of Chemistry,Chemical Engineering Resource Utilization,Northeast Forestry University,Harbin 150040,China;Medical school,Harbin Vocational&Technical College,Harbin 150081,China)

机构地区：[1]东北林业大学生命科学学院,黑龙江哈尔滨150040 [2]东北林业大学化学与资源利用学院,黑龙江哈尔滨150040 [3]哈尔滨职业技术学院医学院,黑龙江哈尔滨150081

出　　处：《食品与发酵工业》2024年第17期198-208,共11页Food and Fermentation Industries

基　　金：黑龙江省自然基金项目-联合引导项目(LH2022C005)。

摘　　要：该文以高温高压超声波辅助法提取白茅根多糖(Rhizoma imperatae polysaccharides,RPS),通过响应面法优化RPS提取工艺。经Sevag法脱蛋白,DEAE-DE纤维素52层析柱和Sephadex G-100凝胶层析柱分离纯化,得到2种RPS(RPS-DS 0和RPS-DS 0.1)。采用α-淀粉酶抑制实验,测定RPS的体外降糖作用。构建胰岛素抵抗HepG2细胞(insulin-resistant HepG2,IR-HepG2)模型,测定RPS-DS 0.1对IR-HepG2细胞中葡萄糖消耗量、糖原含量、己糖激酶和丙酮酸激酶活性的影响。结果表明,在液料比(mL∶g)为20∶1、提取温度112℃、提取时间15 min、超声波功率410 W、超声波处理时间25 min时,RPS质量分数得率为14.79%。RPS-DS 0和RPS-DS 0.1是分子质量分别为179.14 kDa和29.79 kDa的均一中性多糖。RPS-DS 0由阿拉伯糖、半乳糖、葡萄糖、木糖和甘露糖组成,其物质的量比为6.79∶9.68∶75.52∶4.38∶1.88。RPS-DS 0.1由鼠李糖、阿拉伯糖、半乳糖、葡萄糖、木糖、甘露糖和葡萄糖醛酸组成,其物质的量比为2.48∶11.76∶20.08∶50.92∶10.25∶2.08∶2.12。RPS-DS 0和RPS-DS 0.1对α-淀粉酶的半数抑制浓度分别为0.40和0.21 mg/mL。细胞实验结果表明,RPS-DS 0.1能显著提高IR-HepG2细胞中葡萄糖消耗量、糖原含量、己糖激酶和丙酮酸激酶活性,调节糖代谢,从而达到降糖作用。In this study,Rhizoma imperatae polysaccharides(RPS)were extracted using a high-temperature,high-pressure ultrasonic-assisted method.The extraction process of RPS was optimized using response surface methodology.After deproteinization using the Sevag method,two types of polysaccharides from R.imperatae(RPS-DS 0 and RPS-DS 0.1)were obtained through separation and purification using DEAE-DE cellulose 52 chromatography column and Sephadex G-100 gel chromatography column.The hypoglycemic effect of RPS was determined using theα-amylase inhibition test in vitro.The insulin-resistant HepG2(IR-HepG2)cell model was constructed to determine the effects of RPS-DS 0.1 on glucose consumption,glycogen content,hexokinase,and pyruvate kinase activities in IR-HepG2 cells.The RPS mass fraction yield was found to be 14.79%under specific conditions including a liquid-feed ratio of 20∶1(mL∶g),extraction temperature of 112℃,extraction time of 15 min,ultrasonic power of 410 W,and ultrasonic time of 25 min.The neutral polysaccharides RPS-DS 0and RPS-DS 0.1 were found to have molecular weights of 179.14 kDa and 29.79 kDa,respectively,and were homogeneous.RPS-DS 0 consisted of arabinose,galactose,glucose,xylose,and mannose in a molar ratio of 6.79∶9.68∶75.52∶4.38∶1.88.RPS-DS 0.1 consisted of rhamnose,arabinose,galactose,glucose,xylose,mannose,and glucuronic acid in a molar ratio of 2.48∶11.76∶20.08∶50.92∶10.25∶2.08∶2.12.The half maximal inhibitory concentration of RPS-DS 0 and RPS-DS 0.1 onα-amylase were determined to be 0.40 mg/mL and 0.21 mg/mL,respectively.The cellular experiments demonstrated that RPS-DS 0.1 significantly increased glucose consumption,glycogen content and hexokinase levels in IR-HepG2 cells,as well as regulated glucose metabolism,ultimately leading to a hypoglycemic effect.

关 键 词：白茅根多糖 响应面 分离纯化 结构特性 降糖活性 HEPG2细胞 

分 类 号：R284[医药卫生—中药学] R285[医药卫生—中医学]