采用超速离心法从牛乳中分离提取外泌体,通过免疫印迹法、纳米颗粒追踪技术、透射电镜对牛乳外泌体进行鉴定。评价了牛乳外泌体对动物双歧杆菌(Bifidobacterium animalis)F1-3-2生长及益生特性的影响。结果表明,牛乳外泌体促进了动物双歧杆菌F1-3-2的生长,提高了其耐酸、耐胆盐等肠胃环境耐受性,增加了菌体的黏附性。此外,牛乳外泌体增强了动物双歧杆菌F1-3-2的抗炎特性,炎症因子TNF-α、IL-1β水平显著下调。该研究为牛乳外泌体作为新型益生元的应用开发奠定了基础。
Exosomes were extracted from bovine milk by ultracentrifugation. The bovine milk exosomes were identified by immunoblotting, nanoparticle tracking and electron microscopy after extraction. The effects of bovine milk exosomes on the growth and probiotic properties of Bifidobacterium animalis F1-3-2 were evaluated. The results showed that the bovine milk exosomes promoted the growth of B. animalis F1-3-2, improved the gastrointestinal tolerance of B. animalis F1-3-2, such as the acid resistance and the bile salt resistance. Besides, the adhesion of B. animalis F1-3-2 was also promoted. In addition, the bovine milk exosomes induced better anti-inflammatory properties of B. animalis F1-3-2, the levels of inflammatory cytokines like TNF-α, IL-1β significantly decreased. In conclusion, the bovine milk exosomes have the potential to be developed as a novel prebiotic.
[1] COCUCCI E, MELDOLESI J.Ectosomes and exosomes:Shedding the confusion between extracellular vesicles[J].Trends in Cell Biology, 2015, 25(6):364-372.
[2] BARILE L, VASSALLI G.Exosomes:Therapy delivery tools and biomarkers of diseases[J].Pharmacology & Therapeutics, 2017, 174:63-78.
[3] LIAO W, DU Y, ZHANG C H, et al.Exosomes:The next generation of endogenous nanomaterials for advanced drug delivery and therapy[J].Acta Biomaterialia, 2019, 86:1-14.
[4] LIU S R, DA CUNHA A P, REZENDE R M, et al.The host shapes the gut microbiota via fecal microRNA[J].Cell Host & Microbe, 2016, 19(1):32-43.
[5] CHEN I H, XUE L, HSU C C, et al.Phosphoproteins in extracellular vesicles as candidate markers for breast cancer[J].Proceedings of the National Academy of Sciences of the United States of America, 2017, 114(12):3 175-3 180.
[6] PEREIRA P C.Milk nutritional composition and its role in human health[J].Nutrition, 2014, 30(6):619-627.
[7] WALSH C, LANE J A, VAN SINDEREN D, et al.Human milk oligosaccharides:Shaping the infant gut microbiota and supporting health[J].Journal of Functional Foods, 2020, 72:104074.
[8] BETKER J L, ANGLE B M, GRANER M W, et al.The potential of exosomes from cow milk for oral delivery[J].Journal of Pharmaceutical Sciences, 2019, 108(4):1 496-1 505.
[9] ZHOU Q, LI M Z, WANG X Y, et al.Immune-related microRNAs are abundant in breast milk exosomes[J].International Journal of Biological Sciences, 2012, 8(1):118-123.
[10] CHEN T, XIE M Y, SUN J J, et al.Porcine milk-derived exosomes promote proliferation of intestinal epithelial cells[J].Scientific Reports, 2016, 6:33862.
[11] YU S R, ZHAO Z H, XU X Y, et al.Characterization of three different types of extracellular vesicles and their impact on bacterial growth[J].Food Chemistry, 2019, 272:372-378.
[12] TONG L J, HAO H N, ZHANG Z, et al.Milk-derived extracellular vesicles alleviate ulcerative colitis by regulating the gut immunity and reshaping the gut microbiota[J].Theranostics, 2021, 11(17):8 570-8 586.
[13] TENG Y, REN Y, SAYED M, et al.Plant-derived exosomal microRNAs shape the gut microbiota[J].Cell Host & Microbe, 2018, 24(5):637-652.
[14] DAHIYA D K, RENUKA, PUNIYA M, et al.Gut microbiota modulation and its relationship with obesity using prebiotic fibers and probiotics:A review[J].Frontiers in Microbiology, 2017, 8:563.
[15] TURRONI F, MILANI C, VENTURA M, et al.The human gut microbiota during the initial stages of life:Insights from bifidobacteria[J].Current Opinion in Biotechnology, 2022, 73:81-87.
[16] TONG L J, HAO H N, ZHANG X Y, et al.Oral administration of bovine milk-derived extracellular vesicles alters the gut microbiota and enhances intestinal immunity in mice[J].Molecular Nutrition & Food Research, 2020, 64(8):e1901251.
[17] SIDIRA M, KOURKOUTAS Y, KANELLAKI M, et al.In vitro study on the cell adhesion ability of immobilized lactobacilli on natural supports[J].Food Research International, 2015, 76:532-539.
[18] 郝海宁, 仝令君, 张兰威, 等.2种不同牛乳外泌体提取方法的比较研究[J].食品安全质量检测学报, 2020, 11(8):2 569-2 574.
HAO H N, TONG L J, ZHANG L W, et al.Comparative study of 2 kinds of different extraction methods of bovine milk exosomes[J].Journal of Food Safety & Quality, 2020,11(8):2 569-2 574.
[19] WITWER K W, BUZÁS E I, BEMIS L T, et al.Standardization of sample collection, isolation and analysis methods in extracellular vesicle research[J].Journal of Extracellular Vesicles, 2013, 2(1):20360.
