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中国精品科技期刊2020
杨雪丽,牛犇,陈杭君,等. 猴头菇纳米囊泡制备、结构表征及抗氧化活性分析[J]. 食品工业科技,2025,46(2):222−234. doi: 10.13386/j.issn1002-0306.2024030095.
引用本文: 杨雪丽,牛犇,陈杭君,等. 猴头菇纳米囊泡制备、结构表征及抗氧化活性分析[J]. 食品工业科技,2025,46(2):222−234. doi: 10.13386/j.issn1002-0306.2024030095.
YANG Xueli, NIU Ben, CHEN Hangjun, et al. Preparation, Structural Characterization and Antioxidant Activity of Extracellular Nanovesicles from Hericium erinaceus[J]. Science and Technology of Food Industry, 2025, 46(2): 222−234. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2024030095.
Citation: YANG Xueli, NIU Ben, CHEN Hangjun, et al. Preparation, Structural Characterization and Antioxidant Activity of Extracellular Nanovesicles from Hericium erinaceus[J]. Science and Technology of Food Industry, 2025, 46(2): 222−234. (in Chinese with English abstract). doi: 10.13386/j.issn1002-0306.2024030095.

猴头菇纳米囊泡制备、结构表征及抗氧化活性分析

Preparation, Structural Characterization and Antioxidant Activity of Extracellular Nanovesicles from Hericium erinaceus

  • 摘要: 目的:优化猴头菇纳米囊泡的制备方法,考察其抗氧化活性以及对Caco-2细胞氧化损伤的保护作用。方法:采用差速离心与蔗糖密度梯度离心联用法制备猴头菇纳米囊泡(Hericium erinaceus-derived extracellular nanovesicles,HEDENVs),采用单因素实验和响应面试验对纳米囊泡的制备条件进行优化,并用激光纳米粒度仪与透射电镜对其粒径、Zeta电位和形貌进行表征;利用液相色谱质谱联用仪(Liquid chromatography-mass spectrometry,LC-MS)对纳米囊泡进行脂质组学和蛋白质组学分析;通过DPPH、ABTS+和O2自由基清除率评价纳米囊泡的抗氧化活性;H2O2构建Caco-2细胞氧化应激损伤模型,采用激光共聚焦显微镜测定细胞中ROS含量变化。结果:纳米囊泡最佳制备条件:蔗糖浓度25%,离心时间2 h,离心转速86000 r/min,此时纳米囊泡的浓度为402.24 μg/mL;纳米囊泡为具有膜结构的小泡,粒径110.7±16.9 nm,PDI 0.551,Zeta电位−14.2 mV;基于脂质组学和蛋白质组学分析,鉴定出纳米囊泡中大约含有346种脂质与52种蛋白质。体外抗氧化结果表明,400 μg/mL纳米囊泡的DPPH自由基清除率为94.79%、ABTS+自由基清除率为96.67%、O2自由基清除率为96.83%;在H2O2构建的Caco-2细胞氧化损伤模型中,纳米囊泡能显著减少损伤细胞中ROS的产生。结论:体外多种抗氧化评价方法表明纳米囊泡具有一定的抗氧化活性。

     

    Abstract: Objective: To optimize the preparation method of Hericium erinaceus-derived extracellular nanovesicles and investigate their antioxidant activity and protective effect against oxidative damage in Caco-2 cells. Methods: Hericium erinaceus-derived extracellular nanovesicles (HEDENVs) were isolated from fresh Hericium erinaceus by ultracentrifugation and sucrose gradient ultracentrifugation. The optimal extraction process of HEDENVs was optimized by one-way test and response surface test, while the particle size, Zeta potential, and morphology of HEDENVs were characterized by a laser particle size instrument and transmission electron microscope. Lipidomics and proteomics analysis of HEDENVs was performed by liquid chromatography-mass spectrometry. The in vitro antioxidant activity of HEDENVs was evaluated by determining the DPPH, ABTS+, and O2 radical scavenging capacity. A Caco-2 cell oxidative stress damage model was created using H2O2, and changes in ROS content in cells were measured using laser confocal microscopy. Results: The optimal extraction process of HEDENVs was the 25% interface of the sucrose concentration, centrifugation time was 2 h, and centrifugal speed was 86000 r/min, at which the concentration of HEDENVs was 402.24 μg/mL. HEDENVs were nanovesicles with a lipid bilayer membrane, the particle size was 110.7±16.9 nm, PDI=0.551, and the Zeta potential was −14.2 mV. Based on the analysis of lipidomics and proteomics, 346 lipids species and 52 proteins were identified in HEDENVs. The results of in vitro antioxidant test showed that the scavenging rate of 400 μg/mL HEDENVs for DPPH free radical was 94.79%, the scavenging rate for ABTS+ free radical was 96.67%, and the scavenging rate for O2 free radical was 96.83%. An oxidative damage model in Caco-2 cells was established by H2O2, HEDENVs could significantly reduce the production of ROS in damaged cells. Conclusion: It showed that HEDENVs had certain antioxidant activity in various antioxidant experiment.

     

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