多囊卵巢综合征患者高尔基体磷酸化蛋白3表达与颗粒细胞凋亡和促排卵结局的相关性

Relationship of Golgi phosphoprotein-3 expression with granulosa cells apoptosis and controlled ovarian hyperstimulation outcomes patients with polycystic ovary syndrome

目的探讨多囊卵巢综合征(PCOS)患者控制性超促排卵(COH)周期卵巢颗粒细胞高尔基体磷酸化蛋白3(GOLPH3)的表达与颗粒细胞凋亡和COH结局的关系。方法选择因输卵管因素或男方因素首次接受体外受精/卵胞质内单精子显微注射-胚胎移植(IVF/ICSI-ET)助孕治疗的PCOS患者70例和同期非PCOS患者70例作为对照组。应用免疫细胞化学(ICC)和Real-time PCR技术检测颗粒细胞GOLPH3、Bcl-2、Bax、Caspase-3蛋白和mRNA的表达,并行相关性分析。结果①GOLPH3在卵巢颗粒细胞胞质中呈阳性表达,PCOS患者颗粒细胞GOLPH3和Bcl-2的表达低于对照组(P<0.05),Bax和Caspase-3的表达均高于对照组(P<0.05);②PCOS患者卵巢颗粒细胞GOLPH3 mRNA、Bcl-2 mRNA表达水平均低于对照组(P<0.05),Bax mRNA、Caspase-3mRNA表达水平均高于对照组(P<0.05);③PCOS患者中临床妊娠亚组颗粒细胞GOLPH3蛋白和mRNA的表达水平均高于未妊娠亚组(P<0.05);④相关性分析显示:GOLPH3蛋白表达与抗凋亡因子Bcl-2呈正相关(P<0.001),与促凋亡相关因子Bax、Caspase-3呈负相关(P<0.001),与COH结局[获卵数、双原核(2PN)受精数、卵裂数、优质胚胎数、优质囊胚数、可利用胚胎数]呈正相关;颗粒细胞GOLPH3 mRNA表达水平与优质胚胎数呈正相关;多元线性回归分析提示GOLPH3表达与Bcl-2表达、可利用胚胎数呈正相关,与Bax表达呈负相关。结论 COH周期PCOS患者颗粒细胞GOLPH3表达水平下降可能促进颗粒细胞凋亡,影响卵母细胞质量,进而影响胚胎发育潜能。

Objective To explore the effect of Golgi phosphoprotein-3（GOLPH3） in granulosa cells of patients with polycystic ovary syndrome（PCOS） during controlled ovarian hyperstimulation（COH）cycles on granulosa cells apoptosis and COH outcomes. Methods Seventy eligible in vitro fertilization/intracytoplasmic sperm injection-embryo transfer（IVF/ICSI-ET） cases with PCOS owing to the tubal or male factors and 70 corresponding IVF/ICSI-ET cases with non-PCOS were included. Immunocytochemistry and Real-time PCR were used to detect the protein and mRNA expression of GOLPH3, Bcl-2, Bax and Caspase-3 in granulosa cells of the two groups, respectively. Meanwhile, the correlations of GOLPH3 expression with cells apoptosis factors（Bcl-2, Bax and Caspase-3） and COH outcomes were analyzed. Results 1） GOLPH3 protein was detected in the cytoplasm of granulosa cells in two groups. The expressions of GOLPH3 and Bcl-2 protein in PCOS group were lower than those in control group（P〈0.05）, while the expressions of Bax and Caspase-3 protein were higher than those in control group（P〈0.05）. 2） GOLPH3 mRNA and Bcl-2 mRNA in PCOS group were lower than those in control group（P〈0.05）. However, Bax mRNA and Caspase-3 mRNA were higher than those in control group（P〈0.05）. 3） Both the protein and mRNA levels of GOLPH3 expression in pregnant PCOS group were higher than those in non-pregnant PCOS group（P〈0.05）. 4） The expression of GOLPH3 protein in granulosa cells was strongly positively correlated with inhibit apoptosis factor Bcl-2 protein（P〈0.001）, and negatively correlated with pro-apoptosis factors Bax and Caspase-3 protein（P〈0.001）, meanwhile, presented positive correlation with COH outcomes, including the number of retrieved oocytes, two pronucleus（2 PN） fertilizations,cleavages, high-quality embryos, high-quality blastocysts and available embryos. GOLPH3 mRNA was positively correlated with the number of high-quality embryos. Multivariate linear analysis discovered that GOLPH3 expression levels were positively associated with Bcl-2 level and the number of available embryos, while were negatively associated with Bax level. Conclusion Down-expression of GOLPH3 in granulosa cells of PCOS patients during COH period may be associated with the quality of oocytes and the unfavorable embryo developmental potential via promoting granulosa apoptosis.