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宫内注射大肠杆菌脂多糖对患有化脓性阴道分泌物的泽西奶牛子宫 mRNA 基因表达和外周多形核白细胞的影响。

Effects of intrauterine infusion of Escherichia coli lipopolysaccharide on uterine mRNA gene expression and peripheral polymorphonuclear leukocytes in Jersey cows diagnosed with purulent vaginal discharge.

机构信息

Department of Animal Science, University of Minnesota, St. Paul 55108.

Department of Large Animal Clinical Sciences, University of Florida, Gainesville 32608.

出版信息

J Dairy Sci. 2017 Jun;100(6):4784-4796. doi: 10.3168/jds.2016-11643. Epub 2017 Apr 21.

Abstract

The objectives of the current experiment were to investigate the effects of intrauterine treatment of cows with purulent vaginal discharge (PVD) using lipopolysaccharide (LPS) from Escherichia coli on uterine mRNA expression of genes related to inflammatory responses, peripheral polymorphonuclear leukocyte (PMN) function, hematological parameters, and blood concentrations of cortisol, haptoglobin, and progesterone (P4). Jersey cows (n = 3,084) were examined for PVD at 35 ± 6 d postpartum using the Metricheck device (Simcro, Hamilton, New Zealand). At examination, 310 cows had PVD (10.1%), but to ensure proper collection and processing of samples, 267 cows were used in this experiment. Cows were balanced for lactation number, body condition score, and milk yield, and randomly assigned to the control treatment [intrauterine infusion of 20 mL of phosphate-buffered saline (PBS); n = 87] or to receive intrauterine infusion of 20 mL of PBS containing 150 µg (LPS150; n = 91) or 300 µg (LPS300; n = 89) of E. coli LPS. Uteri were biopsied in a subgroup of cows at 6 h after infusion and in another subgroup of cows at 24 h after infusion. Peripheral PMN expression of adhesion molecules (L-selectin and MAC-1) and phagocytosis and oxidative burst were evaluated at 0, 2, and 6 h after infusion. Blood sampled 0, 2, 6, 24, 48, and 168 h after infusion was used for complete hemogram and to determine concentrations of cortisol, haptoglobin, and P4. Treatment did not affect uterine mRNA expression of adhesion molecules [endothelial leukocyte adhesion molecule (E-selectin), intracellular adhesion molecule-1 (ICAM-1), and vascular cell adhesion molecule-1 (VCAM-1)], cytokines (tumor necrosis factor-α, IL-1β, IL-6, IL-8, and IL-10), and toll-like receptor-4. Treatment did not affect PMN expression of L-selectin, but intensity of expression of MAC-1 was higher for LPS150 cows than PBS cows, and tended to be higher in LPS150 than LPS300 cows. Furthermore, a greater percentage of PMN from LPS300 cows were positive for phagocytosis and oxidative burst compared with PBS and LPS150 cows. No effects were observed of treatment on hematological parameters and concentrations of cortisol, haptoglobin, and P4. These observations suggest that intrauterine infusion of E. coli LPS moderately stimulates peripheral PMN function, but further research is needed to better understand the immunomodulatory effects of LPS in the uterus of cows with PVD.

摘要

本实验的目的是研究用大肠杆菌脂多糖(LPS)对患有化脓性阴道分泌物(PVD)的奶牛进行宫内治疗对与炎症反应、外周多形核白细胞(PMN)功能、血液学参数以及皮质醇、触珠蛋白和孕酮(P4)血浓度相关的子宫 mRNA 表达的影响。在产后 35±6d,使用 Metricheck 设备(Simcro,汉密尔顿,新西兰)对 3084 头泽西奶牛进行 PVD 检查。在检查时,有 310 头奶牛患有 PVD(10.1%),但为了确保正确采集和处理样本,本实验仅使用了 267 头奶牛。奶牛根据泌乳次数、体况评分和产奶量进行平衡,并随机分为对照组[宫内输注 20ml 磷酸盐缓冲盐水(PBS);n=87]或接受宫内输注 20ml 含有 150μg(LPS150;n=91)或 300μg(LPS300;n=89)大肠杆菌 LPS 的 PBS。在输注后 6h 对一小部分奶牛的子宫进行活检,并在输注后 24h 对另一部分奶牛的子宫进行活检。在输注后 0、2 和 6h 评估外周 PMN 表达的粘附分子(L-选择素和 MAC-1)和吞噬作用和氧化爆发。在输注后 0、2、6、24、48 和 168h 采血,进行全血细胞计数,并测定皮质醇、触珠蛋白和 P4 的浓度。处理不影响粘附分子[内皮白细胞粘附分子(E-选择素)、细胞间粘附分子-1(ICAM-1)和血管细胞粘附分子-1(VCAM-1)]、细胞因子(肿瘤坏死因子-α、IL-1β、IL-6、IL-8 和 IL-10)和 Toll 样受体-4 的子宫 mRNA 表达。处理不影响 PMN 表达 L-选择素,但 LPS150 奶牛的 MAC-1 表达强度高于 PBS 奶牛,且 LPS150 奶牛的 MAC-1 表达强度高于 LPS300 奶牛。此外,与 PBS 和 LPS150 奶牛相比,LPS300 奶牛的更多 PMN 对吞噬作用和氧化爆发呈阳性。处理对血液学参数以及皮质醇、触珠蛋白和 P4 的浓度没有影响。这些观察结果表明,宫内输注大肠杆菌 LPS 适度刺激外周 PMN 功能,但需要进一步研究以更好地了解 LPS 在患有 PVD 的奶牛子宫中的免疫调节作用。

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