孕激素环境下促排卵对颗粒细胞功能影响的研究

发布时间：2018-03-06 18:41

本文选题：安宫黄体酮　切入点：孕激素环境下促排卵　出处：《山西医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：背景及研究目的:孕激素环境下促排卵(Progestin primed ovarian stimulation,PPOS)是以利用孕激素抑制LH峰为理念的一种新的促排卵方案,临床研究发现黄体期内源性的孕激素及卵泡早期辅助使用外源性孕激素安宫黄体酮(medroxyprogesterone acetate,MPA)均可有效抑制早发LH峰,此外,与短方案相比,PPOS方案需使用较大剂量的人绝经促性腺素(hMG),推测这可能是由于孕激素环境下卵泡对促性腺激素的敏感性略低,亦或是由于孕激素影响了卵泡颗粒细胞的功能而导致的。本课题也验证了卵泡早期使用MPA可以有效抑制早发LH峰这一观点。有学者通过体外培养大鼠颗粒细胞的研究发现,MPA可以抑制雌激素的分泌。体内的研究则发现MPA对促性腺激素诱导的大鼠卵巢分泌的雌激素没有影响,但可抑制卵巢中芳香化酶的活性。因此卵泡早期MPA的使用是否会通过抑制促卵泡生成素(follicle stimulating hormone,FSH)诱导的芳香化酶的表达而影响雌激素的合成,目前尚不清楚。我们的研究目的在于通过检测PPOS方案中卵巢颗粒细胞FSH受体表达水平以了解卵泡对促性腺激素的反应及MPA是否通过影响颗粒细胞分泌雌激素的能力来影响卵泡的生长,旨在为进一步优化促排卵方案奠定基础。方法:本实验选取了2015年3月-2015年10月在上海市第九人民医院辅助生殖中心行体外受精-胚胎移植(in vitro fertilization-embryo transfer,IVF-ET)助孕的不孕症患者共112例。其中行PPOS方案促排卵者62例,50例行短方案促排卵者作为对照组。收集所有患者促排卵过程中血液样本,采用化学发光法检测血清性激素水平。应用密度梯度离心法收集所有患者卵泡液中颗粒细胞,实时荧光定量PCR法(Realtime PCR)和蛋白质印迹法(Western blot)检测颗粒细胞内芳香化酶(CYP19A1)、促性腺激素受体(FSHR、LHR)和雌激素受体(ERβ)的表达水平。结果:1、PPOS组血清LH水平在促排卵过程中逐渐降低,且无一例发生早发LH峰。2、促排卵过程中,血清E2水平随着卵泡的长大逐渐增加,两组间并无统计学差异(P0.05);PPOS组血清FSH水平显著性高于短方案组(P0.05)。3、PPOS组FSH受体(FSHR)在mRNA和蛋白水平均显著高于短方案组FSHR(P0.05),雌激素受体β亚型在蛋白水平明显高于短方案组(P0.05)。4、PPOS组颗粒细胞黄体生成素受体(LHR)和CYP19A1在mRNA水平和蛋白水平与短方案组相比无统计学差异(P0.05)。结论:1、MPA可以有效控制超促排卵中早发LH峰的发生,PPOS可能是全胚冷冻策略下最合适的促排卵方案。2、卵泡期孕激素环境下促排卵对卵巢反应性没有产生不良影响。3、MPA没有对颗粒细胞中芳香化酶的表达及其分泌雌激素的能力产生明显影响。
[Abstract]:Background and objective: Progestin primed ovarian stimulation (PPOSs) is a new ovulatory stimulation scheme using progesterone to inhibit LH peak. Clinical studies have shown that endogenous progesterone in luteal phase and exogenous progesterone medroxyprogesterone acetateMPA) can effectively inhibit early LH peak. Compared with the short regimen, the PPOS regimen requires a higher dose of human menopausal gonadotropin (hMGG), which may be due to the lower sensitivity of follicles to gonadotropin in progesterone environment. Or because progesterone affects the function of follicular granulosa cells. This study also proves that early use of MPA in follicles can effectively inhibit early LH spike. Some scholars in vitro cultured rat granulosa cells. It was found that MPA could inhibit the secretion of estrogen. In vivo, MPA had no effect on gonadotropin-induced estrogen secretion in the ovary of rats. But it can inhibit the activity of aromatase in ovary. Therefore, whether the use of MPA in early follicular stage may affect the synthesis of estrogen by inhibiting the expression of aromatase induced by follicle stimulating hormone. It is not clear yet. Our aim is to investigate the follicle response to gonadotropin by detecting the expression level of FSH receptor in ovarian granulosa cells in PPOS protocol and whether MPA can affect the estrogen secretion of granulosa cells. To affect the growth of follicles, Methods: in order to lay the foundation for further optimization of ovulation promotion program, we selected infertile women who were assisted by IVF in vitro fertilization-embryo transfer IVF-ETS from March 2015 to October 2015 at the assisted Reproductive Center of Shanghai 9th people's Hospital. 62 patients with ovulation induced by PPOS regimen and 50 patients with ovulation induced by short regimen were used as control group. Blood samples were collected from all patients during ovulation promotion. Serum sex hormone levels were detected by chemiluminescence method. Granulosa cells were collected from follicular fluid of all patients by density gradient centrifugation. The expression of aromatase (CYP19A1), gonadotropin receptor (FSHRL) and estrogen receptor ER 尾 (ER 尾) in granulosa cells was detected by real time PCR and Western blot. During ovulation promotion, serum E2 level increased gradually with follicle growth. There was no significant difference between the two groups in the level of serum FSH in P0.05. 3PPOS group. The level of mRNA and protein in mRNA group was significantly higher than that in the short regimen group. The protein level of estrogen receptor 尾 subtype was significantly higher than that in the short regimen group. The level of estrogen receptor 尾 subtype was significantly higher than that in the short regimen group. CYP19A1 and luteinizing hormone receptor (LHRR) in granulosa cells were not significantly different from those in short regimen group in mRNA and protein levels. Conclusion CYP19A1 can effectively control the occurrence of early LH peak in hyperstimulation of ovulation and may be a whole embryo cryopreservation strategy. Ovulatory stimulation did not have any adverse effect on ovarian reactivity under the condition of follicular progesterone. 3 MPA had no significant effect on the expression of aromatase and the ability of estrogen secretion in granulosa cells.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R714.8

【相似文献】