卵巢颗粒细胞高尔基体磷酸化蛋白3表达对颗粒细胞凋亡及卵母细胞发育潜能的影响

Effect of GOLPH3 Expression in Granulosa Cells on the Apoptosis of Ovarian Granulosa Cells and the Developmental Potential of Oocyte

目的:探讨卵巢颗粒细胞高尔基体磷酸化蛋白3(Golgi phosphoprotein 3,GOLPH3)表达对颗粒细胞凋亡及卵母细胞发育潜能的影响。方法:收集2016年5月—2017年4月因输卵管性不孕行体外受精-胚胎移植(IVF-ET)助孕的80例患者的卵巢颗粒细胞,根据妊娠结局分为临床妊娠组40例、未妊娠组40例。采用免疫细胞化学、蛋白质印迹(Western blotting)方法检测2组患者颗粒细胞GOLPH3、Bcl-2、Bax、Caspase-3蛋白的表达,采用实时聚合酶链反应(real-time PCR)方法检测2组患者GOLPH3、Bcl-2、Bax、Caspase-3基因表达,并进行相关性分析。结果:GOLPH3、Bcl-2、Bax、Caspase-3在2组颗粒细胞中均有表达,其中临床妊娠组GOLPH3、Bcl-2的表达高于未妊娠组(P<0.05),而Bax、Caspase-3的表达低于未妊娠组(P<0.05)。相关性分析显示,GOLPH3表达与Bcl-2表达呈显著正相关(P<0.001),与Bax、Caspase-3表达呈显著负相关(均P<0.001),与优质胚胎数、优质囊胚数、可利用胚胎数呈显著正相关(均P<0.001)。结论:卵巢颗粒细胞中GOLPH3表达水平可能通过Bcl-2、Bax、Caspase-3凋亡信号转导通路调控颗粒细胞凋亡作用,并进而对卵母细胞的发育潜能产生影响。

Objective：To evaluate the effect of golgi phosphoprotein 3（GOLPH3） on the apoptosis of granulosa cell and the development potential of oocyte. Methods： A total of 80 women receiving IVF-ET due to tubal factors during the period from May 2016 to April 2017 were enrolled in the study. There were 40 cases in the clinical pregnancy group and 40 cases in the non-pregnant group according to the pregnancy outcome. The expression levels of GOLPH3, Bcl-2, Bax and Caspase-3 protein were determined by immunocytochemistry staining and Western blotting, while the expression levels of m RNAs were quantified by real-time PCR assay.Results： GOLPH3, Bcl-2, Bax and Caspase-3 were all expressed in granulosa cells at various levels. There were significant differences in the expressions of GOLPH3, Bcl-2, Bax and Caspase-3 in granulosa cells between the pregnant group and the non-pregnant group（P〈0.05）. Pearson correlation analysis revealed that the expression of GOLPH3 in granulosa cells was strongly positively correlated with the expression of Bcl-2 protein（P〈0.001）,negatively correlated with the expression of Bax and Caspase-3 protein（both P values 0.001）, and strongly positively correlated with the numbers of high-quality embryo, blastocyst and available embryo（all P〈0.001）.Conclusions： The expression level of GOLPH3 in granulosa cells may play an important role in regulating the apoptosis of granulosa cells through Bcl-2, Bax and Caspase-3 signal pathways, and the developmental potential of oocyte.