食品科学 ›› 2022, Vol. 43 ›› Issue (17): 138-146.doi: 10.7506/spkx1002-6630-20210602-026

• 营养卫生 • 上一篇    下一篇

玉簪多糖对细胞氧化应激损伤的保护作用机制

吴金姗,黄榕,刘树英,刘回民,刘洪章,刘景圣   

  1. (1.吉林农业大学食品科学与工程学院,吉林 长春 130118;2.吉林农业大学生命科学学院,吉林 长春 130118)
  • 出版日期:2022-09-15 发布日期:2022-09-28
  • 基金资助:
    吉林省科技厅项目(20190301046NY);科技部国家成果转化项目(2014GB2B100007)

Protective Mechanism of Polysaccharide from Hosta ventricosa against Oxidative Damage in Cells

WU Jinshan, HUANG Rong, LIU Shuying, LIU Huimin, LIU Hongzhang, LIU Jingsheng   

  1. (1. College of Food Science and Engineering, Jilin Agricultural University, Changchun 130118, China;2. College of Life Science, Jilin Agricultural University, Changchun 130118, China)
  • Online:2022-09-15 Published:2022-09-28

摘要: 本实验以紫萼玉簪(Hosta ventricosa)为原材料,采用响应面法优化其根系多糖(Hosta ventricosa root polysaccharide,HVRP)的提取工艺,探索HVRP对叔丁基过氧化氢(tert-butyl hydroperoxide,t-BHP)诱导的HepG2细胞氧化应激损伤的保护作用机制。结果表明:在提取温度84 ℃、提取时间3 h、料液比1∶31条件下,HVRP平均提取率达到23.9%;HVRP中还原性糖、蛋白质、糖醛酸的质量分数分别为11.17%、0.6%、12.31%,含有多糖类物质的特征吸收峰,不存在三螺旋构象。体外抗氧化和细胞实验结果表明,HVRP具有较强的抗氧化活性,能显著或极显著降低t-BHP诱导氧化损伤HepG2细胞内活性氧、丙二醛含量并提高过氧化氢酶、超氧化物歧化酶、还原型谷胱甘肽及谷胱甘肽过氧化物酶水平(P<0.05、P<0.01)。通过实时荧光定量聚合酶链式反应和蛋白质免疫印迹分析发现,HVRP可通过调控Kelch样环氧氯丙烷相关蛋白1/核因子E2相关因子2/抗氧化反应元件信号通路促进抗氧化酶表达,从而改善t-BHP诱导的HepG2细胞氧化损伤。本研究可为阐明HVRP抗氧化作用机制及其在功能性食品中的应用提供理论依据。

关键词: 紫萼玉簪;多糖;响应面优化;抗氧化活性;细胞损伤

Abstract: In this study, the extraction process of Hosta ventricosa root polysaccharide (HVRP) was optimized by response surface methodology (RSM), and the protective effect of HVRP on oxidative injury induced by tert-butyl hydroperoxide (t-BHP) in HepG2 cells was explored. Under the optimized conditions: extraction temperature 84 ℃, extraction time 3 h and solid-to-liquid ratio 1:31, the average extraction yield of HVRP was 23.9%, The mass fractions of reducing sugar, protein and uronic acid in HVRP were 11.17%, 0.6% and 12.31%, and HVRP showed absorption peaks characteristic of polysaccharides but no triple-helical conformation. HVRP had strong in vitro antioxidant activity and could significantly reduce the contents of reactive oxygen species (ROS) and malondialdehyde (MDA) and increase the activity of catalase (CAT), superoxide dismutase (SOD), reduced glutathione (GSH) and glutathione peroxidase (GPx) (P < 0.05, P < 0.01). The results of real-time quantitative polymerase chain reaction (qPCR) and Western blot analysis showed that HVRP could promotes the expression of antioxidant enzymes and consequently protect HepG2 cells from t-BHP-induced oxidation damage by regulating the Kelch-like epichlorohydrin-associated protein 1/nuclear factor erythroid 2-related factor 2/antioxidant response element (Keap1/Nrf2/ARE) signaling pathway. This study provides a theoretical basis for clarifying the antioxidant mechanism of HVRP and its application in functional foods.

Key words: Hosta ventricosa; polysaccharide; response surface optimization; antioxidant activity; cell damage

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