卵丘颗粒细胞mtDNA拷贝数及4977bp缺失在卵巢储备功能减退中的研究  

Study on mtDNA copy number and 4977 bp deletion in cumulus granulosa cells of diminished ovarian reserve

作　　者：王雪莹 安卓 姚冠峰 郭晓丽 王树松 谢聪聪 WANG Xue-ying;AN Zhuo;YAO Guan-feng;GUO Xiao-li;WANG Shu-song;XIE Cong-cong(Key Reproductive Medical Laboratory of Hebei Province,Key Laboratory for Family Planning and Birth Health of the National Health and Family Planning Committee,Hebei Reproductive Hospital,Hebei Institute of Reproductive Health Science and Technology,Shijiazhuang 050071;Graduate School of Hebei Medical University,Shijiazhuang 050017)

机构地区：[1]河北省生殖健康科学技术研究院河北省生殖医院国家卫生健康委员会计划生育与优生重点实验室/河北省生殖医学重点实验室,石家庄050071 [2]河北医科大学研究生院,石家庄050017

出　　处：《生殖医学杂志》2022年第5期606-612,共7页Journal of Reproductive Medicine

基　　金：河北省医学科学研究课题(20190144)。

摘　　要：目的分析卵巢储备功能减退(DOR)患者卵丘颗粒细胞线粒体DNA(mtDNA)拷贝数和4977 bp缺失情况,初步探讨DOR患者卵丘颗粒细胞mtDNA结构是否完整。方法选取2019年4月至2020年10于河北省生殖医院行IVF/ICSI-ET治疗的不孕症患者33例,根据卵巢储备功能情况分为DOR组(17例),卵巢储备功能正常(NOR)组(16例),比较两组患者的一般情况、实验室指标及妊娠结局。于取卵日挑选患者卵泡液中的卵冠丘复合体后,分别收集每位患者卵母细胞周围的卵丘颗粒细胞团,用线粒体分离试剂盒提取卵丘颗粒细胞的线粒体后提取mtDNA,采用实时定量PCR检测卵丘颗粒细胞mtDNA的拷贝数和4977 bp缺失情况,比较两组患者mtDNA 4977 bp缺失率及拷贝数的差异。结果两组患者的体质量指数(BMI)、基础性激素水平无统计学差异(P>0.05);DOR组的女方年龄显著高于NOR组(P<0.05),DOR组基础窦卵泡数(AFC)显著低于NOR组(P<0.05)。两组患者卵裂率、优质胚胎率、可利用胚胎率、临床妊娠率、种植率均无统计学差异(P>0.05);DOR组促性腺激素(Gn)天数、Gn用量、平均获卵数和受精率显著低于NOR组(P<0.05)。DOR组mtDNA拷贝数(1.04±0.48)略低于NOR组的mtDNA拷贝数(1.13±0.52),但两者尚无显著性差异(P>0.05);且DOR组和NOR组的卵丘颗粒细胞mtDNA均未检测到4977 bp缺失。结论DOR患者卵丘颗粒细胞mtDNA拷贝数并未明显减少,也未见4977 bp片段缺失;其可能与DOR患者临床结局较差并无直接关联。Objective:To analyze mitochondrial DNA(mtDNA) copy number and 4977 bp deletion in cumulus granulosa cells of the patients with diminished ovarian reserve(DOR),and investigate the structure integrity of mtDNA of cumulus granulosa cells in patients with DOR.Methods:Thirty-three infertile patients treated with IVF/ICSI-ET in Hebei Reproductive Hospital from April 2019 to October 2020 were selected. According to the ovarian reserve function, they were divided into DOR group(n=17) and normal ovarian reserve(NOR) group(n=16). The general conditions, laboratory indexes and pregnancy outcomes of the two groups were compared. After the coronoid colliculus complexes were collected from the follicular fluid of the patients on the day of oocytes retrieved, and then the cumulus granulosa cells around oocytes of each patient were collected. Mitochondria of cumulus granulosa cells were extracted with mitochondrial isolation kit, and then mtDNA was extracted. The copy number of mtDNA and 4977 bp deletion of cumulus granulosa cells were detected by real-time quantitative PCR. The deletion rate of mtDNA 4977 bp and mtDNA copy number of cumulus granulosa cells were compared between the two groups.Results:There were no significant differences in body mass index(BMI) and basic sex hormone levels between the two groups(P>0.05). The age of women in DOR group was significantly higher than that in NOR group(P<0.05),and the number of basal antral follicles(AFC) in DOR group was significantly lower than that in NOR group(P<0.05). There were no significant differences in cleavage rate, high-quality embryo rate, available embryo rate, clinical pregnancy rate and implantation rate between the two groups(P>0.05). The days of gonadotropin(Gn) used, the Gn dosage, the average number of oocytes and the fertilization rate in DOR group were significantly lower than those in NOR group(P<0.05). The copy number of mtDNA in DOR group was(1.04±0.48),which was slightly lower than that in NOR Group(1.13±0.52),but there was no significant difference betw

关 键 词：卵巢储备功能 mtDNA拷贝数 4977 bp缺失 

分 类 号：R711.75[医药卫生—妇产科学]