摘要目的 探讨颗粒细胞中胚胎干细胞因子Nanog、Oct4、Sox2与多囊卵巢综合征(PCOS)发生的关系.方法 选择接受体外受精/卵胞质内单精子注射-胚胎移植术(IVF/ICSI-ET)治疗的PCOS患者40例,同期输卵管因素或男方因素不孕患者40例作为对照组.利用Real time-PCR技术检测颗粒细胞中Nanog、Oct4、Sox2以及抗苗勒管激素(AMH) mRNA的表达水平,并用Western blotting分析蛋白表达水平与PCOS发生的关系.结果 PCOS组体质量指数(BMI)[(26.44±3.46) kg/m2]、黄体生成素(LH)[(7.56±0.98) IU/L]、LH/卵泡刺激素(FSH)(1.06±0.15)及获卵数(28.7±3 9)较对照组[(22.43±3.17) kg/m2、(4.26±0.34) IU/L、0.57±0.04、15.1±2 0]有显著升高(P均＜0 05).Real time-PCR结果显示,PCOS组胚胎干细胞因子Nanog (0.60±0.09)和Oct4 (0.85±0.14) mRNA表达水平显著低于对照组(1.65±0.17,1.59±0.17)(P＜0.05),AMH转录水平(1.28±0.11)显著高于对照组(0.89±0.07)(P＜0.05).Logistic同归分析结果显示,Nanog是PCOS发生的危险因素(OR=26.577,P=0.002),且Nanog与BMI、获卵数及AMH呈显著负相关(n=0.415,P=0.023;n=-0.415,P=0.022;r=-0.373,P=0.043),与Oct4呈显著正相关(r=0.684,P＜0.01).Western blotting结果显示,与对照组比较,PCOS组Nanog蛋白表达降低,AMH蛋白表达水平增高.结论 PCOS卵巢颗粒细胞中胚胎干细胞因子Nanog以及AMH的异常表达可能参与了PCOS的发生并影响卵母细胞的发育与成熟,为PCOS的临床治疗提供理论依据.

abstractsObjective To explore the relationship of expressions of embryonic stem cell factor Nanog,Oct4,Sox2ingranulosacellswithpolycysticovarysyndrome(PCOS).Methods Totally 40 PCOS patients and 40 infertile cases due to tubal factors and/or male infertility who underwent in vitro fertilization/intracytoplasmic sperm injectionembryo transfer (IVF/ICSI-ET) at the same period were recruited as experimental group and control group,respectivly.Real-time PCR was used to detect the expression levels of Nanog,Oct4,Sox2 and anti-Müllerian hormone (AMH)mRNA in granulosa cells.Logistic regression analysis was carried out on the pathogenesis of PCOS.Results Body mass index (BMI) [(26.44 ± 3.46) kg/m2],lutenizing hormone (LH) [(7.56 ± 0.98) IU/L],LH/follicluar stimulating hormone (FSH) (1.06 ± 0.15) levels and the number of retrieved oocytes (28.7 ± 3.9) were significantly increased in PCOS group compared with control group [(22.43 ± 3.17) kg/m2,(4.26 ± 0.34) IU/L,0.57 ± 0.04,15.1 ±2.0] (P＜0.05).However,the mRNA levels of Nanog (0.60 ± 0.09) and Oct4 (0.85 ± 0.14) in PCOS group were significantly lower than those in control group (1.65±0.17,1.59±0.17) (P＜0.05) and the mRNA level of AMH (1.28± 0.11) was higher than that in control group (0.89 ± 0.07) (P＜0.05).Logistic regression analysis showed Nanog was a risk factor of PCOS,and the Nanog mRNA level was negatively associated with BMI,number of retrieved oocytes and AMH (r1 =-0.415,P=0.023;r2=-0.415,P=0.022;r3 =-0.373,P=0.043,respectively),and positively correlated with Oct4 (r=0.684,P＜0.01).Western blotting results showed that the Nanog level decreased in PCOS group compared with control group.While,the AMH level increased.Conclusion The abnormal expression levels of Nanog and AMH in granulosa cells may be associated with the morbidity of PCOS and development and mature of oocyte.It will provide theoretical basis for treatment of PCOS.