抗苗勒管激素及卵丘细胞端粒长度与卵母细胞成熟度的相关研究

Relationship between anti-mullerian hormone in follicular fluid and telomere length in cumulus cells with oocytes at different mature stages

目的:探讨体外受精-胚胎移植(in vitro fertilization-embryo transfer,IVF-ET)中卵泡液内抗苗勒管激素(anti-mullerian hormone,AMH)水平及对应的卵丘细胞端粒长度与卵母细胞成熟度的关系。方法:收集52例行IVF-ET患者卵泡液及卵丘细胞,依据每位患者卵母细胞成熟度,将成熟卵泡液、未成熟卵泡液分别混合标记为成熟的卵泡液组(follicular fluids of mature oocyte,FFMⅡ)、未成熟的卵泡液组(follicular fluids of immature oocyte,FFMI)。从卵母细胞周围机械剥离的部分卵丘颗粒细胞同样依据卵母细胞成熟度分组,标记为成熟卵母细胞的颗粒细胞组(cumulus cells of mature oocyte,CCMⅡ)、未成熟卵母细胞的颗粒细胞组(cumulus cells of immature oocyte stage,CCMI)。应用酶联免疫法(enzyme-linked immunosorbent assay,ELISA)检测卵泡液中AMH,实时定量PCR测定卵丘细胞端粒长度。结果:未成熟卵泡液组中AMH水平高于成熟组[(6.02±1.34)vs.(5.66±1.42)],差异未达到统计学意义(P=0.13);未成熟组卵丘细胞端粒长度较成熟组的短[(3.50±1.08)vs.(4.40±1.24)],差异有统计学意义(P=0.000)。结论:未成熟的卵泡液组中AMH水平高于成熟组,卵泡液中AMH可能与卵母细胞成熟度有关,卵丘细胞端粒长度可以反映卵母细胞成熟度。

Objective：To investigate the association between anti-mullerian hormone （AMH） levels in the follicular fluid and corre- sponding cumulus cell telomere length as well as oocyte maturity in vitro fertilization-embryo transfer（IVF-ET）. Methods ：Follicular fluids and cumulus granular cells were obtained from 52 patients during IVF cycles. On the basis of oocyte maturation of each patient, the immature follicle fluid, mature follicle fluid were respectively mixed and marked as mature follicle fluid group （FFM Ⅱ ） and imma- ture follicular fluid（FFMI） group. Cumulus granular cells （CCs） were manually separated from the oocyte-cumulus complex under microscope. All the CCs sample were divided into two groups：granulosa cells of mature oocytes（CCM Ⅱ ） ,granular cell of immature oocytes group（CCMI） based on the maturation of oocyte in every patient. Follicular fluids AMH basal levels were evaluated by ELISA, DNA was extracted from cumulus cells and assessed for telomere length by real-time quantitative PCR. Results：AMH concentrations in the FFMI group were higher than those in FFM Ⅱ group,but without statistical significances[（6.02±1.34） vs. （5.66±1.42） ,P= 0.13]. The telomere length was significantly shorter in CCMI group than in CCM ]I group,with significant differences between the two groups [（3.50 ±1.08）vs. （4.40 ± 1.24）,P=0.000]. Conclusion ：AMH in follicular fluid may reflect the degree of oocyte maturation; the telomere length could be used as a diagnostic criterion for oocyte maturation.