摘要
目的探讨人卵巢黄素化颗粒细胞细胞周期蛋白D2(cyclin D2)的表达与卵巢功能及超排卵过程中卵巢反应性的相关性。方法收集48例行体外受精-胚胎移植(IVF-ET)患者的卵巢黄素化颗粒细胞,根据取卵时卵泡发育数不同将卵巢反应性分为高反应组(12例)、中反应组(30例)及低反应组(6例)。采用免疫细胞化学方法检测颗粒细胞膜上cyclin D2及增殖细胞核抗原(PCNA)蛋白的表达;逆转录聚合酶链反应(RT-PCR)检测颗粒细胞cyclin D2 mRNA的表达水平;化学发光法测定基础血清卵泡刺激素(FSH),人绒毛膜促性腺激素(hCG)日E_2水平。分析颗粒细胞cyclin D2 mRNA及蛋白的表达与IVF临床各项参数、卵巢反应性的关系以及和颗粒细胞增殖活性PCNA标记指数(PCNA-LI)的相关性。结果 (1)黄素化颗粒细胞cyclin D2蛋白表达与年龄、基础FSH水平及促性腺激素(Gn)支数呈负相关(r=0.547,P<0.01;r=-0.456,P<0.05;r=0.451,P<0.05);cyclin D2蛋白表达与获卵数和hCG日E_2水平呈正相关(r=0.592,P<0.01;r=0.626,P<0.01);高、中和低反应三组cyclin D2蛋白表达有显著统计学差异(F=4.995,P<0.05)。(2)黄素化颗粒细胞cyclin D2 mRNA表达与年龄、基础FSH水平呈负相关(r=-0.510,P<0.05;r=-0.891,P<0.01);与获卵数和hCG日E_2水平呈正相关(r=0.629,P<0.01;r=0.524,P<0.05);高、中和低反应三组cyclin D2 mRNA的表达有显著统计学差异(F=8.843,P<0.01)。(3)黄素化颗粒细胞cyclin D2蛋白及mRNA表达与PCNA-LI均呈显著正相关(r=0.696,P<0.01;r=0.498,P<0.05)。结论黄素化颗粒细胞cyclin D2 mRNA及蛋白的表达可反映卵巢储备功能并与卵巢反应性正相关。cyclin D2可能通过调节颗粒细胞的增殖分化参与卵泡的发育。
Objective： To investigate the correlation of cyclin D2 expression in human luteinized granulosa cells with ovarian function and ovarian response. Methods. Ovarian luteinized granulosa cells were collected from 48 women who underwent in vitro fertilization programme in this department. According to the number of follicles ≥16 mm obtained during oocyte retrieval, the patients were divided into three groups： high response group （n= 12）, moderate response group （n=30） and poor response group （n= 6）. Immunocytochemistry was performed to detect the expression of cyclin D2 protein and PCNA protein in human luteinized granulosa cells. The cyclin D2 mRNA in granulosa cells was determined by reverse transcriptase-polymerase chain reaction （RT-PCR） technique. Serum concentrations of hormones were measured by chemil （CLIA）. immunoassay Results： （1）Expression of cyclin D2 protein in human luteinized granulosa cells was negatively correlated with age（r=-0. 547, P〈0.01）, serum baseline FSH （r=-0. 456, P〈0.05） and the ampules of gonadotropins used （r=-0. 451, P〈0.05）, while positively correlated with the number of oocytes retrieved and serum estradiol （E2） levels at the day of hCG administration （r=0. 592, P〈0.01 and r= 0. 626, P〈0.01）, respectively. The expressions of cyclin D2 protein were significantly different among three groups （F=4. 995, P〈0.05）. （2）Expression of cyclin D2 mRNA in human luteinized granulosa cells showed a negative correlation with age （r=-0. 510,P〈0.05） and serum baseline FSH, （r=-0. 891, P〈0. 01）, and a positive correlation with number of oocytes retrieved （r=0. 629, P〈0. 01） and serum Ez levels at the day of hCG administration （r=0. 524, P〈0.05）; cyclin D2 mRNA expression showed a significant difference among different ovarian responders （F = 8. 843, P 〈 0.01）. （3） There was a significant positive correlation of cyclin D2 mRNA （r = 0. 498, P 〈 0.05） and protein expression （r = 0. 696, P〈
出处
《生殖医学杂志》
CAS
2011年第4期313-318,共6页
Journal of Reproductive Medicine
基金
河北省卫生厅科研基金项目(20100055)
