Fiorenza Mariani Farias, Bridi Alessandra, Dos Santos Gislaine, Rosa Paola Maria, Alves Luana, Ferst Juliana Germano, Ferraz Priscila Assis, Pugliesi Guilherme, Pohler Ky, Perecin Felipe, Meirelles Flávio Vieira, da Silveira Juliano Coelho
Departamento de Medicina Veterinária, Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga, SP, Brasil.
Departamento de Reprodução Animal, Faculdade de Medicina Veterinária e Zootecnia, Universidade de São Paulo, Pirassununga, SP, Brasil.
Anim Reprod. 2024 Aug 26;21(3):e20240064. doi: 10.1590/1984-3143-AR2024-0064. eCollection 2024.
This study explored the migration of follicular fluid (FF)-derived extracellular vesicles (EVs) of the uterine environment to the bloodstream and their interaction with neutrophils and . For the experiment, six Nellore heifers () received an intrauterine infusion seven days after ovulation with 1X PBS only (sham group; n=1), 1X PBS stained with lipophilic dye PKH26 (control group; n=2), or FF-derived EVs stained with PKH26 (treated group; n=3). Plasma was collected at 0, 10, 30, 60-, 180-, 360-, 720-, and 1440-min post-infusion to obtained EVs for analysis by nano flow cytometry. Labeled EVs were present in the bloodstream at 30- and 60-min post-infusion in the treatment group. Additionally, plasma derived-EVs from all groups were positive for Calcein-AM, Alix, Syntenin, and Calnexin, which confirm the presence of EVs. The second experiment utilized the plasma-derived EVs from the heifers from 30 and 60 min timepoints to evaluate if neutrophils can uptake EVs . As results, it was possible to observe the presence of labeled EVs in neutrophils treated with plasma derived-EVs from the treatment group. In summary, our results suggest that labeled EVs can migrate from the uterine environment rapidly and interact with circulating immune cells in bovine.
本研究探讨了子宫环境中卵泡液（FF）衍生的细胞外囊泡（EVs）向血液中的迁移及其与中性粒细胞的相互作用。在实验中，六头Nellore小母牛（）在排卵后七天接受子宫内输注，分别输注仅含1X PBS的溶液（假手术组；n = 1）、用亲脂性染料PKH26染色的1X PBS（对照组；n = 2）或用PKH26染色的FF衍生的EVs（处理组；n = 3）。在输注后0、10、30、60、180、360、720和1440分钟采集血浆以获取EVs，用于通过纳米流式细胞术进行分析。在处理组中，输注后30和60分钟时，标记的EVs出现在血液中。此外，所有组的血浆来源的EVs对钙黄绿素-AM、Alix、Syntenin和钙连接蛋白均呈阳性，这证实了EVs的存在。第二个实验利用来自小母牛在30和60分钟时间点的血浆来源的EVs来评估中性粒细胞是否能够摄取EVs。结果发现，在用处理组的血浆来源的EVs处理的中性粒细胞中可以观察到标记的EVs的存在。总之，我们的结果表明，标记的EVs可以迅速从子宫环境迁移并与牛体内循环的免疫细胞相互作用。
