| ObjectThrough detecting the expression of growth differentiation factor-9(GDF-9),bone morphogenetic protein-15(BMP-15)and the expression of apoptosis related factors BCL-2,BAX,we discussed the effect of them on the apotosis of cumulus cells and the oocyte quality of polycystic ovarian syndrome(PCOS)patients undergoing controlled ovarian hyperstimulationon(COH)preliminarily,in order to provide a new thought for improving assisted reproductive techonology outcome of PCOS.MethodsThirty PCOS and sixty control patients who underwent intracytoplasmic sperm injection(ICSI)because their husbands were oligospermatism or asthenospermiain were enrolled in Assisted Reproductive Technology Unit of our hospital from January 2015 to July 2016.All cumulus cells of two groups were collected.Immunocytochemistry was used to detect the expression of GDF-9,BMP-15 protein in cumulus cells and Motic Med 6.0 system was used to analyze average optical density of cumulus cells.The mRNA levels of GDF-9,BMP-15,BAX,BCL-2 in cumulus cells were analyzed by real-time PCR.Cell apotosis was detected by TUNEL and Annexin V-FITC/PI.Using t test to compare measurement data and using chi-square test compare count data between two groups.Using Pearson linear correlation to analyse the relationshiop between two variables.Results1.Age,infertility year,BMI,basal serum FSH,E2 and PRL between the control and PCOS group were no statistically different(P>0.05).Basal LH and T in PCOS were significantly higher than those of control group.After treatment of Diane 35 and metformin,serum FSH,PRL,E2,LH and T between the two groups were no statisticallydifferent when to start COH cycle(P<0.05).The number of punctured follicles,retrieved oocytes,mature oocytes and fertiliazation were higher and the rate of high quality embryo,high quality blastocyst were lower in PCOS than those in control group(P<0.05).2.Immunocytochemistry was used to detect the expression of GDF-9 and BMP-15 protein in cumulus cells whose were positively stained in cumulus cell cytoplasm.Motic Med 6.0 system was used to test protein expression by immunocytochemistry,GDF-9 and BMP-15 levels of PCOS were significantly lower than those of control group(P<0.05)3.GDF-9 and BMP-15 m RNA were analyzed by real-time PCR.GDF-9 and BMP-15 mRNA levels of PCOS were significantly lower than those of control group(P<0.05).4.Apoptosis rate was confirmed by TUNEL and Annexin V-FITC/PI.The apoptosis rate of cumulus cells of PCOS group was higher than those of control group(P<0.05).Then BCL-2、BAX mRNA were also analyzed by real-time PCR,and these expression levels in PCOS were significantly lower than those of control group,but BAX/BCL-2 was significantly higher than which of control group(P<0.05).5.According to Pearson linear correlation,GDF-9 and BMP-15 mRNA level were significantly positively correlated with the rate of normal fertilization,cleavage,high quality embryos.In PCOS group,BCL-2 mRNA level was significantly positively correlated with GDF-9 and BMP-15 m RNA levels,and BAX mRNA level was negatively correlated with GDF-9 and BMP-15 mRNA level(P<0.05).ConclusionleThe results of the study demostated that the rate of normalfertilization,cleavage,high quality embryo and high quality blastocyst of PCOS group was significantly lower than that of control group,the apotosis rate of cumulus cells of PCOS group was also higher than that of control group,which means that the oocyte quality is poorer in PCOS group.GDF-9 and BMP-15 expression levels of PCOS group were significantly lower thanthat of control group,GDF-9 and BMP-15 mRNA levels were significantly positively correlated with BCL-2,negatively correlated with BAX in PCOS group.So we concluded that the lower expression of GDF-9 and BMP-15 mRNA on the ovary cumulus cells in PCOS leaded to the aberrant expression of BCL-2 and BAX mRNA which caused the higher apotosis rate of cumulus cells,this was maybe the reason why poorer oocyte quality in PCOS. |
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