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围产期奶牛亚临床或临床子宫内膜炎时循环和子宫内膜多形核白细胞的活力和功能动态。

Viability and function dynamics of circulating versus endometrial polymorphonuclear leukocytes in postpartum dairy cows with subclinical or clinical endometritis.

机构信息

Department of Internal Medicine, Reproduction and Population Medicine, Faculty of Veterinary Medicine, Ghent University, Salisburylaan, Merelbeke, 9820, Belgium.

Department of Internal Medicine, Reproduction and Population Medicine, Faculty of Veterinary Medicine, Ghent University, Salisburylaan, Merelbeke, 9820, Belgium; Laboratory of Veterinary Physiology and Biochemistry, Department of Veterinary Sciences, University of Antwerp, Universiteitsplein, Wilrijk, 2610, Belgium.

出版信息

J Dairy Sci. 2023 May;106(5):3436-3447. doi: 10.3168/jds.2022-22471. Epub 2023 Mar 17.

Abstract

We aimed to compare the viability of circulating polymorphonuclear leukocyte (cPMN) and endometrial PMN (ePMN) and their function dynamics in postpartum dairy cows with subclinical (SCE) or clinical endometritis (CE). To do so, blood samples from 38 Holstein cows were collected at -7, 9, 21, and 36 d relative to calving, and endometrial cytology samples from 32 Holstein cows were harvested at 9, 21, and 36 d postpartum. Uterine health status was assessed at 36 d postpartum, and cows were classified as healthy (absence of abnormal vaginal discharge and ≤5% ePMN), SCE (absence of abnormal vaginal discharge and >5% ePMN), or CE (mucopurulent or purulent vaginal discharge and >5% ePMN). Viability (viable, apoptotic, and necrotic) and function parameters phagocytosis (PC), oxidative burst, and intracellular proteolytic degradation were evaluated for cPMN via flow cytometry. For ePMN, only viability and PC were evaluated. The association of cPMN and ePMN viability and functional parameters with reproductive tract health classification were fitted in mixed linear regression models, accounting for repeated measures, sampling day, and interactions of reproductive tract status and day. Cows with CE had a lower proportion of cPMN viability (84.5 ± 2.1%; least squares means ± standard error) and a higher proportion of apoptosis (14.4 ± 2.0%) than healthy (92.4 ± 1.3 and 6.7 ± 1.3%, respectively) or SCE (95.3 ± 2.4 and 3.8 ± 2.3%, respectively) at 9 d postpartum. Interestingly, cPMN intracellular proteolytic degradation was lower [6.2 ± 0.1 median fluorescence intensity (MFI)] in SCE compared with healthy (6.7 ± 0.08 MFI) or CE (6.8 ± 0.1 MFI) at d 9 postpartum. No other differences in cPMN function were found among experimental groups. The proportion of necrotic ePMN was higher for healthy (49.6 ± 5.1%) than SCE (27.4 ± 7.3%) and CE (27.7 ± 7.3%) cows at 36 d postpartum. Also, at 36 d postpartum, the proportion of ePMN performing PC was higher in CE (47.0 ± 8.6%) than in healthy (18.4 ± 7.6%) cows, but did not differ from SCE cows (25.9 ± 8.7%). Results of the present study suggest that cPMN viability and function at 9 d postpartum are associated with the development of uterine disease. Furthermore, ePMN at 36 d postpartum are mostly necrotic in healthy cows but viable and functional in cows with CE, probably due to active uterine inflammation. Remarkably, ePMN in cows with SCE at 36 d postpartum are also mostly viable but seem to display a numerically lower proportion of PC compared with ePMN in CE cows.

摘要

我们旨在比较患有亚临床(SCE)或临床子宫内膜炎(CE)的产后奶牛循环多形核白细胞(cPMN)和子宫内膜多形核白细胞(ePMN)的活力及其功能动态。为此,在产后第-7、9、21 和 36 天采集了 38 头荷斯坦奶牛的血液样本,并在产后第 9、21 和 36 天采集了 32 头荷斯坦奶牛的子宫内膜细胞学样本。在产后 36 天评估子宫健康状况,将奶牛分为健康(无异常阴道分泌物且 ePMN 不超过 5%)、SCE(无异常阴道分泌物且 ePMN 超过 5%)或 CE(黏液脓性或脓性阴道分泌物且 ePMN 超过 5%)。通过流式细胞术评估 cPMN 的活力(存活、凋亡和坏死)和吞噬作用(PC)、氧化爆发和细胞内蛋白水解降解等功能参数。对于 ePMN,仅评估活力和 PC。将 cPMN 和 ePMN 活力和功能参数与生殖道健康分类的相关性拟合到混合线性回归模型中,该模型考虑了重复测量、采样日以及生殖道状态和日的相互作用。患有 CE 的奶牛在产后第 9 天的 cPMN 活力(84.5 ± 2.1%;最小二乘均值 ± 标准误差)较低,凋亡比例(14.4 ± 2.0%)较高,而健康(92.4 ± 1.3% 和 6.7 ± 1.3%)或 SCE(95.3 ± 2.4% 和 3.8 ± 2.3%)。有趣的是,产后第 9 天,SCE 奶牛的 cPMN 细胞内蛋白水解降解水平低于健康奶牛(6.2 ± 0.1 中位荧光强度(MFI))和 CE 奶牛(6.7 ± 0.08 MFI)。在实验组之间未发现 cPMN 功能的其他差异。产后第 36 天,健康奶牛(49.6 ± 5.1%)的坏死 ePMN 比例高于 SCE(27.4 ± 7.3%)和 CE(27.7 ± 7.3%)奶牛。此外,产后第 36 天,CE 奶牛的 ePMN 执行 PC 的比例高于健康奶牛(47.0 ± 8.6%),但与 SCE 奶牛(25.9 ± 8.7%)没有差异。本研究的结果表明,产后第 9 天 cPMN 的活力和功能与子宫疾病的发展有关。此外,产后第 36 天健康奶牛的 ePMN 主要是坏死的,但 CE 奶牛的 ePMN 具有活力和功能,可能是由于子宫炎症活跃。值得注意的是,产后第 36 天 SCE 奶牛的 ePMN 也主要是有活力的,但与 CE 奶牛的 ePMN 相比,它们似乎表现出较低的 PC 比例。

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