食品工业科技2025，Vol.46Issue(2)：218-230,13.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
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 ultracen-trifugation 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.关键词
猴头菇/纳米囊泡/工艺优化/结构表征/组学分析/Caco-2细胞/抗氧化Key words
Hericium erinaceus/nanovesicles/process optimization/structural characterization/omics analysis/Caco-2 cells/antioxidation分类
轻工业引用本文复制引用
杨雪丽,牛犇,陈杭君,吴伟杰,王冠楠,房祥军,穆宏磊,郜海燕..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):218-230,13.基金项目
国家自然科学基金(32172282) （32172282）
国家重点研发计划课题(2021YFD2100502) （2021YFD2100502）
浙江省高层次人才特殊支持计划(2022R52025). （2022R52025）
