Lin Dian-Liang, Quan Song, Kang Yue-Fan, Yu Ai-Li, Lin Yuan
Fujian Provincial Reproductive Medicine Center, FujianMaternity and Children's Hospital Affiliated to Fujian Medical University, Fuzhou 350001, China. E-mail:
Nan Fang Yi Ke Da Xue Xue Bao. 2017 Oct 20;37(10):1351-1357. doi: 10.3969/j.issn.1673-4254.2017.10.11.
To evaluate the impact of GOLPH3 expression in cumulus granulosa cells on the outcomes of intracytoplasmic sperm injection (ICSI).
A total of 119 women receiving ICSI due to male infertility at our center between April, 2012 and June, 2014 were enrolled in the study. Cumulus granulosa cells were collected from the women for detection of GOLPH3 expressions using immunocytochemistry, Western blotting, and real-time PCR. GOLPH3 expression rate was compared between women with and without clinical pregnancy following ICSI, and the associations of GOLPH3 expression with the laboratory indicators of ICSI outcomes were assessed.
Immunocytochemistry showed that GOLPH3 expression was located mainly in in the plasma of the cumulus granulosa cells. The rate and intensity of GOLPH3 expression in the cumulus granulosa cells differed significantly between women with and without clinical pregnancy following ICSI (P<0.05). GOLPH3 expression was found to positively correlate with the numbers of punctured follicles, grade III oocyte cumulus complex, ICSI oocytes, fertilized oocytes, cleavage, high quality embryos, blastocysts, high quality blastocysts, and frozen embryos (all P<0.01). The results of RTPCR and Western blotting revealed significant differences in GOLPH3 expressions at both the mRNA and protein levels in the cumulus granulosa cells between the pregnant and non-pregnant groups after ICSI (t=14.560, P=0.000). Western blot analysis revealed significant difference of GOLPH3 protein expression in cumulus granulosa cells between women with and without clinical pregnancy following ICSI.
GOLPH3 expression in the cumulus granulosa cells plays an important role in the development of oocytes and promotion of conception to affect the outcomes of ICSI.
评估卵丘颗粒细胞中GOLPH3表达对卵胞浆内单精子注射(ICSI)结局的影响。
选取2012年4月至2014年6月期间在本中心因男性不育接受ICSI的119名女性纳入研究。收集这些女性的卵丘颗粒细胞,采用免疫细胞化学、蛋白质印迹法和实时荧光定量PCR检测GOLPH3表达。比较ICSI后临床妊娠和未妊娠女性的GOLPH3表达率,并评估GOLPH3表达与ICSI结局实验室指标的相关性。
免疫细胞化学显示,GOLPH3表达主要位于卵丘颗粒细胞的胞浆中。ICSI后临床妊娠和未妊娠女性卵丘颗粒细胞中GOLPH3表达的率和强度差异有统计学意义(P<0.05)。发现GOLPH3表达与穿刺卵泡数、III级卵母细胞-卵丘复合体、ICSI卵母细胞、受精卵母细胞、卵裂、优质胚胎、囊胚、优质囊胚及冷冻胚胎数量均呈正相关(均P<0.01)。实时荧光定量PCR和蛋白质印迹法结果显示,ICSI后妊娠组与未妊娠组卵丘颗粒细胞中GOLPH3在mRNA和蛋白质水平的表达差异有统计学意义(t=14.560,P=0.000)。蛋白质印迹分析显示,ICSI后临床妊娠和未妊娠女性卵丘颗粒细胞中GOLPH3蛋白表达差异有统计学意义。
卵丘颗粒细胞中GOLPH3表达在卵母细胞发育及促进受孕过程中起重要作用,从而影响ICSI结局。