University of Queensland Centre for Clinical Research, The University of Queensland, Brisbane, Queensland, 4029, Australia.
Cognosco, Anexa FVC, Morrinsville 3340, New Zealand.
J Dairy Sci. 2017 Nov;100(11):9143-9152. doi: 10.3168/jds.2017-13261. Epub 2017 Aug 31.
A contributing factor to declining fertility in dairy cows is an activated inflammatory system associated with uterine infection. Detecting uterine disease using biomarkers may allow earlier diagnosis and intervention with resultant improvements in fertility. Exosomes are known to participate in intercellular communication, paracrine, and endocrine signaling. Exosomes carry a cargo of proteins, lipids, and nucleic acids that represent specific cellular sources. Prostaglandins are lipids that are critical determinants of bovine fertility. In this study exosomes were isolated from the plasma of cows before (d 0) and during (d 10) the study in healthy animals or those with an induced uterine infection in a 2 × 2 factorial design. Exosomes were characterized for size and number (nanoparticle tracking analysis), exosomal marker expression (Western blot), and morphology (transmission electron microscopy). No significant differences were observed in exosome size or number. The abundance of exosome-enriched markers was confirmed in noninfected and infected animals. Transmission electron microscopy confirmed the morphology of the exosomes. These exosomes were co-incubated with bovine endometrial epithelial and stromal cells. Exosomes from d-10-infected animal plasma decreased PGF production in endometrial epithelial but not stromal cells. For future research, the identification of effectors in the cargo may provide a useful basis for early diagnosis of uterine infection using an exosomal characterization approach.
奶牛生育率下降的一个促成因素是与子宫感染相关的激活炎症系统。使用生物标志物检测子宫疾病可以更早地诊断和干预，从而提高生育率。外泌体已知参与细胞间通讯、旁分泌和内分泌信号传递。外泌体携带蛋白质、脂质和核酸等货物，这些货物代表特定的细胞来源。前列腺素是脂质，是牛生育力的关键决定因素。在这项研究中，在健康动物或在 2×2 析因设计中诱导子宫感染的动物中，在研究开始前（d0）和期间（d10）从血浆中分离外泌体。通过纳米颗粒跟踪分析对大小和数量进行了特征分析，通过 Western blot 对外泌体标记物的表达进行了分析，通过透射电子显微镜对形态进行了分析。在外泌体大小或数量方面未观察到显著差异。在未感染和感染动物中均证实了富含外泌体的标记物的丰度。透射电子显微镜证实了外泌体的形态。将这些外泌体与牛子宫内膜上皮和基质细胞共孵育。来自 d10 感染动物血浆的外泌体减少了子宫内膜上皮细胞而不是基质细胞中的 PGF 产生。对于未来的研究，对货物中效应物的鉴定可能为使用外泌体特征分析方法早期诊断子宫感染提供有用的基础。
