目的:在孕激素引发的卵巢刺激(PPOS)周期中,醋酸甲羟孕酮(MPA)的暴露是否会引起颗粒细胞类固醇生成功能和促性腺激素反应性的分子扰动?
结论:PPOS周期与传统的GnRH拮抗剂周期相同,不仅在临床IVF特征上,而且在促性腺激素表达上也在对促性腺激素的反应,以及分子水平的类固醇生成功能。
背景:由于孕酮通过降低下丘脑水平的GnRH搏动性对LH释放的抑制作用,因此PPOS越来越多地用作GnRH拮抗剂的替代品。尽管越来越多的临床研究证据表明PPOS和拮抗剂方案在IVF周期特征和产科结局方面没有显着差异,尚不清楚在卵巢刺激期间,腔卵泡队列暴露于孕酮或其合成衍生物是否会导致类固醇生成和促性腺激素反应性方面的任何细微的分子异常.为了解决这个问题,我们对接受PPOS和拮抗剂周期的正常反应IVF患者获得的叶黄素化的壁颗粒细胞(GC)进行了详细的分子比较分析.
方法:对IVF患者进行临床转化研究。
方法:本研究包括55名正常反应的IVF患者,他们接受了使用MPA(5mg,每日两次)或GnRH拮抗剂醋酸西曲瑞克的PPOS刺激卵巢。FSH和hCG的重组形式用于卵巢刺激和排卵触发,分别。在卵母细胞回收过程中获得的黄素化壁GC用于实验。细胞培养,实时定量PCR,免疫印迹,共聚焦延时活细胞成像,和激素检测。
结果:接受PPOS和GnRH拮抗剂卵巢刺激的患者的人口统计学和IVF周期特征相似,包括卵巢反应,成熟卵母细胞产量,和受精率。分子分析显示,参与性类固醇合成的酶的表达(StAR,SCC,3β-HSD,17β-HSD,芳香化酶)和胆固醇(LDL受体,激素敏感性脂肪酶,羟甲基戊二酰辅酶A还原酶,PPOS循环的GC和甾醇O-酰基转移酶1)与拮抗剂循环的GC相当。促性腺激素受体的表达,雌激素,孕激素也相似。与拮抗剂周期的GC相比,基础和hCG诱导的3β-HSD表达和孕酮产生的增加以及基础和FSH诱导的PPOS患者GC的芳香化酶表达和E2输出的增加没有任何有意义的差异。此外,基础和hCG诱导的LDL受体表达和胆固醇摄取上调在两组之间没有差异.共聚焦成像还揭示了类固醇生成酶的相似表达模式及其与线粒体的共定位。最后,调节积云扩张的其他重要基因的表达,排卵,和黄体功能[松弛素,PPOS和拮抗剂周期的GC中的ADAMTS-1和表皮生长因子(EGF)样生长因子双调蛋白]相似。
方法:不适用。
结论:在解释我们的数据时,应谨慎行事,这些数据来自正常反应患者,其排卵是由hCG触发的。目前尚不清楚评估的分子参数是否因不孕症病因而异,卵巢反应的大小,触发模式,和任何其他潜在的卵巢病变或全身性疾病。MPA是用于PPOS的孕激素,这些发现是否可以推广到其他孕激素尚不清楚。
结论:这项研究提供了令人放心的分子证据,表明在卵泡生长期,腔卵泡群暴露于MPA不会对类固醇生成产生任何有害影响,排卵,与GnRH拮抗剂周期相比,黄体功能。
背景:这项研究由医学院资助,Koc大学健康科学研究生院和Koc大学转化医学研究中心(KUTTAM),同样由土耳其共和国发展研究基础设施支持计划部资助。所有作者都声明没有利益冲突。
背景:不适用。
OBJECTIVE: Does medroxyprogesterone acetate (MPA) exposure in progestin-primed ovarian stimulation (PPOS) cycles cause molecular perturbations in the steroidogenic function and gonadotropin responsiveness of the granulosa cells?
CONCLUSIONS: PPOS cycles are identical to traditional GnRH antagonist cycles not only for clinical IVF characteristics but also for gonadotropin receptor expression, response to gonadotropins, and steroidogenic function at the molecular level.
BACKGROUND: PPOS is increasingly used as an alternative to GnRH antagonists due to the inhibitory effect of progesterone on LH release by reducing GnRH pulsatility at the hypothalamic level. Although a growing body of evidence from clinical studies did not indicate significant differences between PPOS and antagonist protocols for IVF cycle characteristics and obstetrical outcomes, it is still unknown whether exposure of the antral follicle cohort to progesterone or its synthetic derivatives during ovarian stimulation causes any subtle molecular aberrations in terms of
steroidogenesis and gonadotropin responsiveness. To address this issue, detailed comparative molecular analyses were conducted in the luteinized mural granulosa cells (GCs) obtained from normal responding IVF patients undergoing PPOS and antagonist cycles.
METHODS: A clinical translational research study was conducted with IVF patients.
METHODS: This study included 55 normal responding IVF patients who underwent ovarian stimulation with either PPOS using MPA (5 mg twice daily) or GnRH antagonist cetrorelix acetate. Recombinant forms of FSH and hCG were used for ovarian stimulation and ovulation triggering, respectively. Luteinized mural GCs obtained during the oocyte retrieval procedure were used for the experiments. Cell culture, quantitative real-time PCR, immunoblotting, confocal time-lapse live cell imaging, and hormone assays were used.
RESULTS: Demographic and IVF cycle characteristics of the patients undergoing ovarian stimulation with PPOS and GnRH antagonist were similar, including ovarian response, mature oocyte yield, and fertilization rates. Molecular analyses revealed that the expression of the enzymes involved in sex-steroid synthesis (StAR, SCC, 3β-HSD, 17β-HSD, aromatase) and the uptake/storage/utilization of cholesterol (LDL receptor, Hormone-sensitive lipase, hydroxy-methyl glutaryl Co-enzyme-A reductase, and Sterol O-acyltransferase1) in the GCs of the PPOS cycles were comparable to those of the antagonist cycles. The expression of the receptors for gonadotropins, estrogen, and progesterone hormones was also similar. Basal and hCG-induced increases in 3β-HSD expression and progesterone production and basal and FSH-induced increases in aromatase expression and E2 output of the GCs from PPOS patients did not exhibit any meaningful differences when compared with GCs from antagonist cycles. Furthermore, basal and hCG-induced up-regulation in the LDL receptor expression and cholesterol uptake did not differ between the groups. Confocal imaging also revealed similar patterns of expression for the steroidogenic enzymes and their co-localization with mitochondria. Lastly, the expression of the other important genes regulating cumulus expansion, ovulation, and luteal function [Relaxin, ADAMTS-1, and epidermal growth factor (EGF)-like growth factor amphiregulin] in the GCs of the PPOS and antagonist cycles were similar.
METHODS: N/A.
CONCLUSIONS: Caution should be exercised when interpreting our data which was derived from normally responding patients whose ovulation was triggered with hCG. It is unclear whether the molecular parameters assessed vary according to infertility etiologies, magnitude of ovarian response, mode of trigger, and any other underlying ovarian pathologies or systemic diseases. MPA was the progestin used for PPOS and whether these findings can be generalized to other progestins is unknown.
CONCLUSIONS: This study provides reassuring molecular evidence that exposure of antral follicle cohorts to MPA during the follicular growth phase does not have any detrimental effects on steroidogenic, ovulatory, and luteal functions when compared with GnRH antagonist cycles.
BACKGROUND: This study was funded by the School of Medicine, the Graduate School of Health Sciences of Koc University and Koç University Research Center for Translational Medicine (KUTTAM), and equally funded by the Republic of Turkey Ministry of Development Research Infrastructure Support Program. All authors declare no conflict of interest.
BACKGROUND: N/A.