Abstract
Objective
To evaluate the composition of bioactive substances and the antioxidant effects of jujube fruit under gut microbiota fermentation (GMF), and the inhibitory effect on cytotoxicity caused by ethyl carbamate (EC).
Methods
Changes in the contents of flavonoids, polyphenols, total sugars, and reducing sugars of jujube fruit after GMF (0, 2, 6, 12, 24, and 48 h) were determined. The oxidation resistance of fermented jujube fruits (from 0 to 48 h fermentation) was evaluated using in vitro 2,2′-azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) and ferric reducing antioxidant power (FRAP) assays. Inhibitory effects of 48 h-fermented jujube fruit at various concentrations (0.25, 0.50, 1.00, and 2.00 mg/mL) on EC-treated toxicity and DNA damage of Caco-2 cells were estimated using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and nuclear staining assays, respectively. Effects of different concentrations of jujube fruit on EC-treated Caco-2 cells’ intracellular reactive oxygen species (ROS), glutathione (GSH) levels, and mitochondrial membrane potential (MMP) were also evaluated.
Results
Jujube fruit has rich bioactive components after GMF and shows strong antioxidant capacity. Fermented jujube fruit can inhibit the cytotoxicity and DNA damage of Caco-2 cells caused by EC and reduce intracellular ROS generation, as well as restoring GSH and MMP.
Conclusions
Fermented jujube fruit extracts produced by GMF still contain biologically active substances which retain biological activity and antioxidation capabilities.
摘要
目的
研究红枣在肠道菌群酵解作用下的生物活性物质组成和抗氧化功效, 以及其对氨基甲酸乙酯诱导细胞毒性的保护作用, 为红枣在肠道菌群酵解后发挥抗氧化作用提供新思路。
创新点
首次研究了肠道菌群酵解对红枣中生物活性物质组成和抗氧化能力的影响及其对氨基甲酸乙酯诱导细胞毒性的保护作用。
方法
红枣模拟肠道菌群酵解 (0∼48h) 后, 采用比色法和高效液相色谱法分析黄酮, 多酚, 总糖和还原糖在发酵过程中的含量变化; 采用体外抗氧化活性评价方法研究红枣发酵代谢产物抗氧化活性; 采用MTT法和 Hoechst 33258 荧光探针研究不同浓度 (0.25, 0.50, 1.00 和 2.00 mg/mL) 红枣发酵48 h代谢产物对氨基甲酸乙酯诱导 Caco-2 细胞的细胞活力和DNA损伤的影响; 分别采用 6-羧基-2′,7′-二氯二氢荧光素二乙酸酯 (DCFH-DA) 和萘-2,3-二甲醛 (NDA) 荧光探针检测不同浓度 (0.25, 0.50 和 1.00 mg/mL) 红枣发酵 48 h代谢产物对氨基甲酸乙酯诱导 Caco-2 细胞内活性氧 (ROS) 和谷胱甘肽 (GSH) 水平; 采用 RH123 荧光探针检测不同浓度 (0.25, 0.50和1.00 mg/mL) 红枣发酵48 h代谢产物对氨基甲酸乙酯诱导 Caco-2 细胞内线粒体膜电位的影响。
结论
红枣经肠道菌群酵解后含有丰富的生物活性成分, 具有很强的抗氧化能力。 红枣发酵代谢物可抑制氨基甲酸乙酯诱导的Caco-2细胞活力下降和DNA损伤, 减少细胞内ROS生成, 恢复GSH水平和线粒体膜电位, 抑制氨基甲酸乙酯诱导的细胞氧化损伤。
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Acknowledgments
This work was supported by the National Natural Science Foundation of China (Nos. U1703105 and 21876152) and the Lishui Science and Technology Bureau Research Fund (No. 2020zdhz01), China.
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Tao BAO performed fermentation experiments, HPLC analysis, in vitro antioxidant activity and cell experiments, and wrote the initial manuscript. Ming ZHANG performed in vitro antioxidant activity experiments and wrote the initial manuscript. Yuanqing ZHOU performed fermentation experiments and HPLC analysis. Wei CHEN conceived the project, designed the study, and supervised the experiments. All authors have read and approved the final manuscript. Therefore, all authors have full access to all the data in the study and take responsibility for its integrity and security.
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Tao BAO, Ming ZHANG, Yuanqing ZHOU, and Wei CHEN declare that they have no conflict of interest.
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Bao, T., Zhang, M., Zhou, Y. et al. Phenolic profile of jujube fruit subjected to gut microbiota fermentation and its antioxidant potential against ethyl carbamate-induced oxidative damage. J. Zhejiang Univ. Sci. B 22, 397–409 (2021). https://doi.org/10.1631/jzus.B2000754
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DOI: https://doi.org/10.1631/jzus.B2000754