Winters Kathleen R H, Meyer Evelyne, Van Merris Valérie M, Van Den Broeck Wim L M, Duchateau Luc, Burvenich Christian
Department of Physiology, Biochemistry and Biometrics, Faculty of Veterinary Medicine, Milk Secretion and Mastitis Research Center, Ghent University, Salisburylaan 133, B-9820, Merelbeke, Belgium.
Steroids. 2003 May;68(5):397-406. doi: 10.1016/s0039-128x(03)00040-0.
During the periparturient period, dairy cows are subjected to physiological changes that may induce immunosuppression and an increased susceptibility of the animal to bacterial infections such as mastitis. The incidence of clinical environmental mastitis is high during the last period of gestation, at parturition and during the first month of lactation, suggesting a potential influence of sex steroid hormones. Efficient functioning of polymorphonuclear leukocytes (PMN) is necessary during the early phase of infection to clear the mammary gland from invading pathogens. The purpose of this study was to investigate the effect of sex steroid hormones on the oxidative burst activity of isolated PMN from ovariectomized cows. Ovariectomy was performed to minimize the interference of endogenous estrogen and progesterone levels, which are known to vary extensively during the estrus cycle. Isolated PMN were incubated with different concentrations of 17beta-estradiol, estrone or progesterone. A flow cytometric technique was used to quantify the oxidation of intracellular 2',7'-dichlorofluorescin by the oxidative burst system of PMN following stimulation with phorbol myristate acetate. Staurosporine was used as a positive control for our in vitro model. No statistically significant changes in PMN oxidative burst activity were observed at physiological or pharmacological levels of the three sex steroid hormones. A large variation existed in the oxidative burst activity among cows. In an additional experiment, the expression of estrogen receptor alpha and of progesterone receptor in PMN was evaluated immunohistochemically. No specific staining was detected for both receptors in isolated PMN following incubation with different concentrations of sex steroid hormones.
在围产期,奶牛会经历一些生理变化,这些变化可能会导致免疫抑制,并增加动物对诸如乳腺炎等细菌感染的易感性。临床环境性乳腺炎的发病率在妊娠后期、分娩时以及泌乳的第一个月较高,这表明性类固醇激素可能具有潜在影响。在感染早期,多形核白细胞(PMN)的有效功能对于清除乳腺中的入侵病原体至关重要。本研究的目的是调查性类固醇激素对去卵巢奶牛分离出的PMN氧化爆发活性的影响。进行卵巢切除术以尽量减少内源性雌激素和孕激素水平的干扰,已知这些激素在发情周期中会有很大变化。将分离出的PMN与不同浓度的17β-雌二醇、雌酮或孕酮一起孵育。使用流式细胞术技术来量化在用佛波酯肉豆蔻酸酯刺激后PMN氧化爆发系统对细胞内2',7'-二氯荧光素的氧化。星形孢菌素用作我们体外模型的阳性对照。在三种性类固醇激素的生理或药理水平下,未观察到PMN氧化爆发活性有统计学上的显著变化。奶牛之间的氧化爆发活性存在很大差异。在另一项实验中,通过免疫组织化学评估了PMN中雌激素受体α和孕激素受体的表达。在用不同浓度的性类固醇激素孵育后,在分离出的PMN中未检测到这两种受体的特异性染色。