目的 探讨卵源性因子——生长分化因子9(GDF9)和骨形态发生蛋白15(BMP15)在多囊卵巢综合征(PCOS)患者及因男方因素不孕的卵巢功能正常妇女刺激周期卵母细胞不同成熟阶段的表达变化及意义.方法 收集12例PCOS患者(PCOS组)的25个卵母细胞[9个胚泡(CV)期,9个MⅠ期和7个M Ⅱ期]和56例卵巢功能正常妇女(对照组)的82个卵母细胞(29个GV期,26个MⅠ期和27个MⅡ期),采用巢式实时定量PCR技术,检测两组妇女不同成熟阶段的卵母细胞中GDF9和BMP15 mRNA的表达水平.结果(1)GDF9 mRNA的表达水平:PCOS组GV、MⅠ、MⅡ期卵母细胞GDF9 mRNA的表达水平分别为44.8(4.2 ~529.0)、27.6(9.8~172.7)和49.0(0.2~65.9),对照组GV、MⅠ、MⅡ期卵母细胞GDF9 mRNA的表达水平分别为149.9(55.4 ~ 387.9)、29.9(2.5 ~205.8)和657.8(149.4 ~1376.2),两组妇女MⅡ期卵母细胞GDF9 mRNA的表达水平分别比较,差异有统计学意义(P<0.01),但两组妇女GV、MⅠ期卵母细胞GDF9 mRNA的表达水平比较,差异无统计学意义(P>0.05);PCOS组不同成熟阶段卵母细胞GDF9 mRNA的表达水平比较,差异无统计学意义(P>0.05),对照组不同成熟阶段卵母细胞GDF9 mRNA的表达水平分别比较,差异均有统计学意义(P <0.05、P<0.01).(2)BMP15 mRNA的表达水平:PCOS组GV、MⅠ、MⅡ期卵母细胞BMP15 mRNA的表达水平分别为0.1(0.1 ~22.0)、3.2(0.6~55.0)和6.4(3.2~8.5),对照组GV、MⅠ、MⅡ期卵母细胞BMP15 mRNA的表达水平分别为41.6(6.5 ~96.1)、4.0(2.0~10.4)和49.7(2.3 ~139.5),两组妇女GV期卵母细胞BMP15 mRNA的表达比较,差异有统计学意义(P<0.01),但两组MⅠ、MⅡ期卵母细胞BMP15 mRNA的表达水平比较,差异均无统计学意义(P>0.05);PCOS组不同成熟阶段卵母细胞BMP15 mRNA的表达水平比较,差异无统计学意义(P>0.05),对照组不同成熟阶段卵母?
Objective To investigate the expression pattern and significance of two importantoocyte-secreted factors:growth differentiation factor 9(GDF9)and bone morphogenetic protein 15(BMP15)during oocyte maturation in women with polycystic ovary syndrome(PCOS)and infertile women due to husband factors.Methods Total of 25 oocytes[9 at germinal vesicle GV stage,9 at M Ⅰ stage and 7 at M Ⅱ stage]were obtained from 12 patients with PCOS and 82 oocytes(29 at GV stage,26 at M Ⅰ stage and 27 at M Ⅱ stage)were from 56 controls.The nested quantitative real time(RT)PCR was uscd to detect the abundance of GDF9 and BMP15 mRNA in each oocyte.Results(1)The expression level of GDF9 mRNA at the GV stage,M Ⅰ stage and M Ⅱ stage in PCOS group were 44.8(4.2-529.0),27.6(9.8-172.7)and 49.0(0.2-65.9)respectively,the expression in were 149.9(55.4-387.9),29.9(2.5-205.8)and 657.8(149.4-1376.2)in control group,respectively.The expression of GDF9 mRNA at M Ⅱ stage was significantly lower in PCOS group than in controls(P 〈 0.01),however,the differences didn't reach statistical significant at GV or M Ⅰ stage between the two groups(P 〉 0.05).The expression of GDF9 mRNA displayed some changes at different maturation stage in controls(P 〈 0.05,P 〈 0.01),however,the expression didn't demonstrate any dynamic changes in PCOS group(P 〉 0.05).(2)The expression level BMP15 mRNA at the GV stage,M Ⅰ stage and M Ⅱ stage in PCOS group were 0.1(0.1-22.0),3.2(0.6-55.0)and 6.4(3.2-8.5),respectively,the expression were 41.6(6.5-96.1),4.0(2.0-10.4)and 49.7(2.3-139.5)in control group,respectively.The expression of BMP15 mRNA at GV stage was significantly lower in PCOS group than in controls(P 〈 0.01),however,the differences were not significant at M Ⅰ or M Ⅱ stage between the two groups(P 〉0.05).The expression of BMP15 mRNA also displayed some changes at different maturation stage in controls(P 〈 0.05),however,the level