枸杞多糖的分离纯化及其对H2O2损伤的HepG2细胞的抗氧化机制研究

Isolation and Purification of Lycium barbarum Polysaccharide and Its Antioxidant Mechanism on HepG2 Cells Damaged by H2O2

关键词:  枸杞多糖  单糖构成  HepG2细胞  抗氧化能力  Nrf2/HO-1通路

Key words:  Lycium barbarum polysaccharide  Monosaccharide composition  HepG2 cells  Antioxidant capacity  Nrf2/HO-1 pathway

摘要:

目的：本研究旨对枸杞粗多糖进行进一步纯化，初步分析其化学结构组成，并通过细胞实验研究枸杞多糖的抗氧化机制。方法：本研究采用超声辅助法提取枸杞多糖并通过DEAE-52纤维素柱进行纯化。使用高效凝胶渗透色谱、傅立叶变换红外光谱和离子色谱分析其结构构成。通过DPPH，ABTs，FRAP和CUPRAC四种方法对枸杞多糖组分进行初步的体外抗氧化评价。利用H2O2诱导HepG2细胞模型，评价枸杞多糖组分抗氧化活性。结果：本研究共取得3种枸杞多糖组分：LBP-C1，LBP-C2和LBP-C3。结构分析发现3种枸杞多糖组分存在不同的分子量，官能团与单糖构成。体外抗氧化研究表明，LBP-C1具有最好的抗氧化潜力。细胞实验发现LBP-C1能通过调节Nrf2/HO-1通路，减轻HepG2细胞氧化应激状态。结论：枸杞多糖LBP-C1具有较高的抗氧化潜力，可应用于后续抗氧化功能药品或食品开发研究。

Abstract:

This study aims to further purify the crude polysaccharides of Lycium barbarum L., preliminarily analyze their chemical structure and composition, and study the antioxidant mechanism of L. barbarum polysaccharides through cell experiments. METHODS This study used ultrasound assisted extraction of L. barbarum polysaccharides and purified them using DEAE-52 cellulose column. Their structures were analyzed by high performance gel permeation chromatography, Fourier transform infrared spectroscopy and ion chromatography. Preliminary in vitro antioxidant evaluation of L. barbarum polysaccharides was conducted using four methods: DPPH, ABTs, FRAP, and CUPRAC. Evaluate the antioxidant activity of L. barbarum polysaccharides using H2O2 induced HepG2 cell model. RESULTS This study obtained a total of three components of L. barbarum polysaccharide: LBP-C1, LBP-C2, and LBP-C3. Structural analysis revealed that three types of L. barbarum polysaccharide have different molecular weights, functional groups, and monosaccharides. In vitro antioxidant studies had shown that LBP-C1 has the best antioxidant potential. Cell experiments had found that LBP-C1 can alleviate the oxidative stress state of HepG2 cells by regulating the Nrf2/HO-1 pathway. CONCLUSION LBP-C1, a polysaccharide of LBP, has high antioxidant potential, which can be applied to the subsequent research of antioxidant function medicine or food development.