Novel sampling procedure to characterize bovine subclinical endometritis by uterine secretions and tissue
Novel sampling procedure to characterize bovine subclinical endometritis by uterine secretions and tissue
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DOI：
10.1016/j.theriogenology.2019.09.016
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发表时间：
2020-01-01
期刊：
影响因子：
2.8
通讯作者：
Zerbe, Holm
中科院分区：
文献类型：
--
作者：
Helfrich, Anika L.;Reichenbach, Horst-Dieter;Zerbe, Holm
牛的亚临床子宫内膜炎（SE）定义为临床上不明显的子宫内膜炎症。据报道，它会损害受影响奶牛的生育能力，并在乳制品行业造成经济损失。诊断SE的金标准尚未确定。子宫细胞学和组织病理学都被应用，但这些方法之间的一致性较低。本研究的目的是评估子宫分泌物（US）作为诊断SE的新介质的能力。应用一种新的取样工具，在108头奶牛（产后43-62天[dpp]）单次通过子宫颈后，检索子宫内膜的US以及细胞学、组织学和细菌学样本。为了评估US样本的质量，对来自5名健康供体的样本进行了蛋白质组分析，证明US的体内采样是可行的，并生成了适用于诊断目的的样本。综合临床、细胞学和组织病理学检查，诊断SE。使用AlphaLISA技术对美国的促炎和抗炎细胞因子（白细胞介素（IL）1B、IL 6、IL 8、IL 17 A、IL 10）进行定量分析。RNA酶固定的子宫内膜活检组织用于细胞因子IL 1B、IL 6、IL 8、IL 10和肿瘤坏死因子α（TNF α）以及胰高血糖素-内过氧化物合酶2（PTGS 2）和抗菌肽S100 A9的基因表达分析，通过逆转录定量实时聚合酶链反应（RT-qPCR）。根据奶牛的子宫健康状况将其分组。一大组动物（n = 83）未显示尿道炎体征（E.NEG）。细胞病理学和组织病理学检查显示一致性较低;因此，将SE动物分为SE（细胞）和SE（组织）组（分别为n = 7和n = 13）。从进一步分析中排除了SE组（细胞+组织）中的1只动物以及4只具有临床结肠炎（CE）体征的动物。与E.NEG.相比，SE（细胞）显示US中IL 1B、IL 8和IL 17 A的中位浓度显著更高，以及子宫内膜活检中IL 1B、IL 8和IL 10的中位表达显著更高。子宫内膜组织中IL 6、TNF α、PTGS 2和S100 A9在US中无显著差异。与E. NEG相比，SE（组织学）在分析参数方面无差异。本研究成功地建立了一种检测奶牛超声的方法。细胞因子IL 1B、IL 8和IL 17 A是超声诊断细胞性腹膜炎的理想候选细胞因子。US的进一步评估可能有助于更好地了解导致慢性子宫内膜炎症和受影响奶牛生育力受损的病理机制。(C)2019爱思唯尔公司All rights reserved.
Subclinical endometritis (SE) in cattle is defined as clinically unapparent inflammation of the endometrium. It is reported to impair fertility in affected cows and causes economic loss within the dairy industry. A gold standard for diagnosis of SE has not been set. Uterine cytology and histopathology are both applied, but low agreement between these methods has been described. The objective of the present study was to assess the capability of uterine secretions (US) as a new medium for diagnosis of SE. A novel sampling tool was applied to retrieve US as well as cytological, histological and bacteriological samples of the endometrium after a singular passage through the cervix in 108 dairy cows (43-62 days post-partum [dpp]). To assess the quality of the US samples, a proteome analysis of samples from five healthy donors was performed, demonstrating that in vivo sampling of US was feasible and generated samples suitable for diagnostic purposes. Diagnosis of SE was realized by the combination of clinical, cytological, and histopathological findings. Quantitative analysis of pro- and anti-inflammatory cytokines (interleukin (IL) 1B, IL6, IL8, IL17A, IL10) in US was conducted using AlphaLISA-technology. RNAlater-fixed endometrial biopsies were used for gene expression analysis of the cytokines IL1B, IL6, IL8, IL10 and tumor necrosis factor alpha (TNF alpha) as well as the prostaglandin-endoperoxide synthase 2 (PTGS2) and the antimicrobial peptide S100A9 by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR). Cows were assigned to groups according to their uterine health status. A large group of animals (n = 83) displayed no signs of endometritis (E.NEG). Cytological and histopathological examination revealed low agreement; hence, animals with SE were differentiated into SE(cyto) and SE(histo) groups (n = 7 and n = 13, respectively). One animal in group SE(cyto + histo) as well as four animals with signs of clinical endometritis (CE) were excluded from further analysis. SE(cyto) showed significantly higher median concentrations of IL1B, IL8 and IL17A in US as well as a significantly higher median expression of IL1B, IL8 and IL10 in endometrial biopsies compared to E.NEG. No significant differences were found for IL6 and IL10 in US and IL6, TNF alpha, PTGS2 and S100A9 in endometrial tissue between these groups. SE(histo) presented no differences concerning the analyzed parameters compared to E.NEG. In conclusion, a method to sample US was successfully established in dairy cows. The cytokines IL1B, IL8 and IL17A are promising candidates in diagnosing cytological endometritis by US. Further assessment of US might contribute to a better understanding of the pathological mechanisms leading to chronic endometrial inflammation and to impaired fertility in affected cows. (C) 2019 Elsevier Inc. All rights reserved.