摘要
目的探讨囊性纤维化跨膜转导调节因子(cystic fibrosis transmembrane conductance regulator,CFTR)在不同年龄不孕患者卵泡液中表达的变化与卵子及胚胎发育结局的相关性。方法采用全自动电化学发光仪测定血清中卵泡刺激素(follicle-stimulating hormone,FSH),黄体生成素(luteotropic hormone,LH),雌二醇(estradiol,E2),孕激素(progesterone,P)水平;采用双抗体夹心ABC-ELISA法测定卵泡液中CFTR的浓度;收集取卵日卵丘颗粒细胞,采用间接免疫荧光染色法检测卵丘颗粒细胞上CFTR蛋白的定位及表达; Real-time PCR测定卵丘颗粒细胞CFTR mRNA的表达。结果 CFTR在不同年龄段不孕患者卵泡液中均有表达,不同年龄段患者卵泡液中CFTR浓度差异有统计学意义(P<0. 05),卵泡液中CFTR水平与年龄呈明显正相关(r=0. 497,P<0. 05)。不同年龄组胚胎培养结局中获卵数和优质胚胎率与卵泡液中CFTR含量呈反比。不同年龄组胚胎培养结局中2PN形成率与卵泡液中CFTR含量呈正比。免疫荧光染色表明颗粒细胞胞浆和胞膜上均有CFTR蛋白的表达。不同年龄患者的卵丘颗粒细胞上均有一定量的CFTR mRNA表达;不同年龄患者卵丘颗粒细胞中CFTR mRNA表达与年龄呈正相关(r=0. 440)。不同年龄可用胚胎率与CFTR mRNA表达量呈反比趋势。结论 CFTR可能参与人卵泡正常生长的调控,与不孕患者年龄有密切的相关性,直接影响获卵数、2PN形成率、优质胚胎率等卵子及胚胎的发育结局,CFTR有可能作为预测体外受精患者卵子及胚胎发育结局的一种无创、定量的标记分子。
Objective To investigate the relationship between cystic fibrosis transmembrane conductance regulator（CFTR） level in follicular fluid and embryo development outcome in infertile women undergoing IVF/ICSI-ET.Methods Serum follicle-stimulating hormone（FSH）, luteotropic hormone（LH）, estradiol（E 2）, progesterone（P） were determined by electrochemiluminescence method. The concentration of CFTR in follicular fluid was detected by double antibody ABC-ELISA. The oval granulosa cells were collected on the day of ova harvest. The expression of CFTR protein in granular cells was detected by indirect immunofluorescence staining, and the expression of CFTR mRNA was detected by RT-PCR.Results There was significant difference in concentration of CFTR in the follicular fluid among women with different ages（ P 〈0.05）; the level of CFTR in follicular fluid was positively correlated with age（ r =0.497, P 〈0.05）. The number of acquired eggs and high quality embryo rate in the embryo culture of different age groups was negatively correlated with the CFTR concentration in the follicular fluid. The rate of 2PN formation in the embryo culture was correlated with CFTR concentration in follicular fluid. The immunofluorescence staining showed that CFTR protein was expressed in the cytoplasm and membrane of granulosa cells. There was a trend of positive correlation between expression of CFTR mRNA in the cumulus granulosa cells with the age of patients（ r =0.440, P =0.560）. There was also a trend of negative correlation between number of retrieved eggs, available embryos and the age of patients; and between embryo ratio and CFTR mRNA expression.Conclusion CFTR levels in follicular fluid are closely related to the age of women with infertility. CFTR may be involved in modulating normal human follicles growth, affecting oocyte retrieval and the number of oocytes, 2PN formation rate, high-quality embryo rate. The expression of CFTR may be used as an indicator to predict the outcome of oocyte and embryo dev
作者
韦慧
王炎秋
张迅轶
纪亚忠
WEI Hui;WANG Yan qiu;ZHANG Xun yi;JI Ya zhong(Dept.of Reproductive Medicine,Tongji Hospital,Tongji University,Shanghai 200065,China)
出处
《同济大学学报（医学版）》
CAS
2018年第5期60-65,共6页
Journal of Tongji University(Medical Science)
基金
上海市科委重点实验室项目(17DZ2273600)
