Department of Obstetrics and Gynecology, Ankara University, Faculty of Veterinary Medicine, Ankara, Turkey.
Department of Obstetrics and Gynecology, Kırıkkale University, Faculty of Veterinary Medicine, Kırıkkale, Turkey.
Domest Anim Endocrinol. 2020 Jan;70:106398. doi: 10.1016/j.domaniend.2019.106398. Epub 2019 Sep 17.
The aim of this study was to investigate some of the growth and transcriptional factors originating from oocytes and granulosa cells in follicular fluid and to identify the relationships between the basic blood metabolite-metabolic hormones and intrafollicular lipopolysaccharide (LPS) concentrations. Thirty cows included in the study were allocated into 2 groups comprising 15 cows with healthy preovulatory follicles (cyclic cows) and 15 cows with confirmed cystic follicles. The ovaries and uteri of all cows were assessed by transrectal ultrasonographic examination. Blood serum samples were collected at 15, 25, 35, 45, and 55 d after calving for analysis of nonesterified fatty acids, β-hydroxybutyrate, insulin, glucose, IGF-I, ACTH, and cortisol. Ovaries and uteri were examined using transrectal ultrasound. Vaginal discharge was evaluated on the same days. Follicular fluid was also aspirated on days 35-55 from the healthy preovulatory follicles and cystic follicles using a transvaginal ovum pickup method. The densitometric levels of inhibin-α, growth and differentiation factor (GDF-9), bone morphogenetic protein (BMP-6), and GATA-4 and GATA-6 proteins were analyzed by the Western blotting technique; the concentrations of antimullerian hormone (AMH), IGF-I, estradiol-17 beta (E2), and progesterone (P4) were determined by ELISA; and the concentrations of LPS in the follicular fluid were measured by the Limulus amebocyte lysate test. The serum insulin, ACTH, and cortisol concentrations were higher in cystic cows than cyclic cows, but serum IGF-I concentrations were lower in cystic cows. The IGF-I concentrations of cystic follicular fluids were lower, whereas AMH levels were significantly greater than those of healthy preovulatory follicular fluids. The cystic follicles had significantly lower expression levels of GDF-9, BMP-6, GATA-4, and GATA-6; in contrast, inhibin-α expression and LPS concentrations were significantly higher than in healthy preovulatory follicles. The proportion of pathologic vaginal discharge within 25 d postpartum in cystic cows were higher than in the cyclic group. In conclusion, it is suggested that intrafollicular dysregulation of the transforming growth factor-β superfamily, growth, and transcriptional factors is affected by high intrafollicular LPS concentrations and systemic metabolic changes and these disturbances may be responsible for the generation of ovarian cysts.
本研究旨在探讨卵泡液中来源于卵母细胞和颗粒细胞的一些生长和转录因子，并确定基础血液代谢物-代谢激素与卵泡内脂多糖（LPS）浓度之间的关系。将 30 头奶牛分为两组，每组 15 头，一组为有健康排卵前卵泡（周期性奶牛），另一组为有明确的囊性卵泡。所有奶牛的卵巢和子宫均通过直肠超声检查进行评估。产后 15、25、35、45 和 55 天采集血样，用于分析非酯化脂肪酸、β-羟丁酸、胰岛素、葡萄糖、IGF-I、ACTH 和皮质醇。使用直肠超声检查卵巢和子宫。同日评估阴道分泌物。使用经阴道取卵法，在第 35-55 天从健康排卵前卵泡和囊性卵泡中抽吸卵泡液。通过 Western 印迹技术分析抑制素-α、生长分化因子（GDF-9）、骨形态发生蛋白（BMP-6）和 GATA-4 和 GATA-6 蛋白的密度水平；通过 ELISA 测定抗缪勒管激素（AMH）、IGF-I、雌二醇-17β（E2）和孕酮（P4）的浓度；通过鲎试验测量卵泡液中的 LPS 浓度。囊性奶牛的血清胰岛素、ACTH 和皮质醇浓度高于周期性奶牛，而血清 IGF-I 浓度低于囊性奶牛。囊性卵泡液中的 IGF-I 浓度较低，而 AMH 水平明显高于健康排卵前卵泡液。囊性卵泡的 GDF-9、BMP-6、GATA-4 和 GATA-6 的表达水平明显较低；相反，抑制素-α的表达和 LPS 浓度明显高于健康排卵前卵泡。产后 25 天内囊性奶牛的病理性阴道分泌物比例高于周期性奶牛组。总之，提示卵泡内转化生长因子-β超家族、生长和转录因子的失调受到卵泡内 LPS 浓度和全身代谢变化的影响，这些干扰可能是卵巢囊肿产生的原因。
