多囊卵巢综合征患者卵母细胞超微结构观察

Ultrastructure of oocytes in women with polycystic ovary syndrome

目的对多囊卵巢综合征(PCOS)患者卵母细胞的超微结构和线粒体DNA(mt DNA)拷贝数进行分析,探讨PCOS患者卵母细胞质量下降的原因。方法根据临床指征,选取2017年1～12月于中国科学技术大学附属第一医院生殖医学中心因男方因素行卵胞质内单精子注射技术(ICSI)助孕的患者115例,根据是否患有PCOS分为试验组和对照组,其中试验组为PCOS患者57例,对照组为非PCOS患者58例。所有患者均接受标准长方案控制性促排卵,共收集187枚卵母细胞用于透射电镜(TEM)的超微结构分析和mt DNA拷贝数的荧光实时定量PCR检测。其中试验组52枚用于超微结构观察,另38枚用于mt DNA拷贝数检测;对照组57枚和40枚分别用于超微结构观察和mt DNA拷贝数检测。对比观察试验组与对照组来源的卵母细胞超微结构各指标:透明带、卵周隙、微绒毛、皮质颗粒、高尔基体和内质网的形态和分布、线粒体的结构与数量。结果与对照组相比,试验组卵母细胞的透明带、卵周隙、微绒毛、皮质颗粒、高尔基体、内质网等细胞结构和细胞器没有变化。但试验组卵母细胞内异常线粒体比例(58. 7%高于对照组17. 7%(P <0. 05),且线粒体数量试验组(44. 3±9. 6)个/切片低于对照组(65. 4±12. 7)个/切片(P <0. 05)。同时,试验组卵母细胞所含的mt DNA拷贝数为(54 390±19 139)个,低于对照组的(88 135±44 235)个(P <0. 05)。结论 PCOS患者卵母细胞内线粒体结构和数量均出现改变,可能与其卵母细胞质量受损有关。

Objective To analyze the ultrastructural and Mitochondria DNA copies number of oocytes from women with polycystic ovary syndrome in vitro fertilization- embryo transfer, and to explore the reasons for the decline of oocyte quality in PCOS patients. Methods According to clinical data, 115 women, who received ICSI cycles due to male factors, were enrolled in this study, including 57 PCOS patients and 58 age-matched controls. All patients received controlled ovarian stimulation using standard long stimulation protocol, and 187 oocytes in total were collected for ultrastructural analysis using transmission electron microscopy and detection of mtDNA copies number using real time PCR. From PCOS oocytes, 52 were used for ultrastructural observation, and another 38 were used for mtDNA copy number detection. Meanwhile, 57 were used for ultrastructural observation and 40 for mtDNA copy number detection respectively in control oocytes. Ultrastructure of oocytes from the test and the control group： zona pellucida, perivitelline space, microvilli, cortical granules, Golgi body and endoplasmic reticulum morphology and distribution, mitochondria structure and quantity. Results No difference was detected in the ultrastructure of zona pellucida, perivitelline space, microvilli, cortical granules, Golgi body and endoplasmic reticulum in PCOS and control oocytes. However, the percentage of abnormal mitochondria in the cytoplasm of PCOS oocytes increased significantly （PCOS oocytes 58.7% vs control oocytes 17.7%, P 〈0.05）, while the total number of mitochondria decreased significantly （PCOS oocytes 44.3±9.6/section vs control oocytes 65.4±12.7/section, P 〈0.05）. Compared with control oocytes, mtDNA copies of PCOS oocytes decreased （PCOS oocytes54390 ± 19139 vs control oocytes 88135 ± 44235, P 〈0.05） . Conclusion The alteration of percentage of abnormal mitochondria and number of mitochondria in PCOS patients might contribute to poor oocyte quality.