FSHR基因多态性在颗粒细胞中的表达与卵巢储备功能及妊娠结局的相关研究

Relationship between expression of FSHR gene polymorphism in granulosa cells and ovarian reserve function and pregnancy outcome

目的探讨卵泡刺激素受体(FSHR)基因多态性在颗粒细胞中的表达与卵巢储备功能及临床治疗结局的相关性,为辅助生殖技术(ART)助孕提供个体化治疗方案。方法以2020年9月至2022年10月于内蒙古医科大学附属医院生殖医学中心接受ART治疗的103例不孕症女性为研究对象。取卵后,提取废弃卵泡液中颗粒细胞的RNA,反转录获取cDNA,聚合酶链反应(PCR)扩增后,基因测序患者FSHR-rs6166的单核苷酸多态性(SNP)位点基因型。根据FSHR-rs6166 SNP位点基因测序结果将患者分为AA组(野生型,n=51)、AG组(杂合型,n=40)及GG组(突变型,n=12),比较各组患者的一般资料、卵巢储备功能、妊娠结局及卵巢的反应性。结果纳入的103例行ART助孕的患者中,FSHR-rs6166基因型以AA型最常见,频率为49.5%。GG组基础卵泡刺激素(bFSH)值[(6.90±1.30)U/L]及AG组bFSH值[(6.26±1.54)U/L]显著高于AA组[(5.65±0.90)U/L,P=0.003],GG组的抗苗勒管激素(AMH)水平[(3.20±1.28)ng/ml]及AG组AMH水平[(3.36±1.62)ng/ml]显著低于AA组[(4.29±1.97)ng/ml,P=0.036];3组患者间的窦卵泡数(AFC)、促性腺激素(Gn)启动量、Gn总量、Gn天数、人绒毛膜促性腺激素(HCG)日雌二醇(E 2)、获卵数、MⅡ卵数、2PN数、卵裂数、受精率、优质胚胎率、临床妊娠率、活产率、卵巢反应性差异均无统计学意义(P>0.05)。结论突变型及杂合型FSHR-rs6166位点可能与女性卵巢储备功能下降有一定关联,但FSHR-rs6166基因型对卵巢的反应性和妊娠结局影响并不明显。

Objective:To investigate the relationship between the expression of FSH receptor(FSHR)gene polymorphism in granulosa cells and ovarian reserve function and clinical outcome,and to provide individualized treatment for assisted reproductive technology(ART).Methods:A total of 103 infertile women who were treated with ART at the Reproductive Medicine Center of the Affiliated Hospital of Inner Mongolia Medical University from September 2020 to October 2022 were selected as the subjects.After oocytes retrieval,the RNA of granulosa cells in abandoned follicular fluid was extracted,and cDNA was obtained by reverse transcription.After polymerase chain reaction(PCR)amplification,the single-nucleotide polymorphism(SNP)locus genotype of FSHR-rs6166 was sequenced.According to the results of FSHR-rs6166 polymorphism gene sequencing,the patients were divided into AA group(wild type)(n=51),AG group(heterozygous type)(n=40)and GG group(mutant type)(n=12).The general data,ovarian reserve function,pregnancy outcome and ovarian responsiveness of patients in each group were compared.Results:Among 103 women with ART,AA genotype was the most common among the three groups of FSHR-rs6166 genotypes,with the frequency of 49.5%.The basal FSH(bFSH)levels in the GG group[(6.90±1.30)U/L]and the AG group[(6.26±1.54)U/L]were significantly higher than those in the AA group[(5.65±0.90)U/L](P=0.003),while the levels of anti-Mullerian hormone(AMH)in the GG group[(3.20±1.28)ng/ml]and the AG group[(3.36±1.62)ng/ml]were significantly lower than those in the AA group[(4.29±1.97)ng/ml](P=0.036).There were no significant differences in the antral follicles count(AFC),initial dose of gonadotropin(Gn),total dose of Gn,number of Gn days,estradiol(E 2)on HCG day,number of oocytes retrieved,number of MⅡoocytes,number of 2PN,number of cleavage,fertilization rate,high-quality embryos rate,clinical pregnancy rate,live birth rate,and ovarian responsiveness among the three groups(P>0.05).Conclusions:The mutant type and heterozygous type at FSHR-rs6166 locus may be associated with a decrease in female ovarian reserve function.The impact of three genotypes on ovarian responsiveness and pregnancy outcomes is not significant.