Study Of The Relationship Between AMH And IVF-ET Outcome In PCOS Patients

In present,In-vitro fertilization and Embryo transfer(IVF-ET) has been broadly used in treatment of sterility.One of the focuses is to find out the factors predicting the result and pregnancy rate during the treatment of IVF.Among these factors,how to acquire high quality oocyte and embryo is the most important factor,which determines the success of IVF-ET and is also one the difficult point during operation. The quality of oocyte is associated with various factors during the generation, development,growth and maturation of ovarian follicle.In recent years,more and more attention has been paid to the effect of local micro-regulation of ovary on the growth and development of follicle;together with other factors,it regulates the orderly consecutive development procedure of follicle from primordial follicle, primary follicle,and secondary follicle to mature dominant follicle.Anti-m(u|¨)llerian hormone(AMH) secreted by granular cells is a member of superfamily of transforming growth factor,which is also called m(u|¨)llerian inhibiting substance (MIS).In 1947,a study conducted by Jost confirmed that a factor differenced from testosterone might induce the degeneration of m(u|¨)llerian duct during the development of male embryo,i.e.anti-m(u|¨)llerian hormone.AMH is a glycoprotein dimer composed of two same 70 kb monomers.During the development of male embryo,the mechanism of the degeneration of m(u|¨)llerian duct is the secretion of AMH,which results in the normal development of male reproductive duct.In female,m(u|¨)llerian duct is the initial cells forming uterus,uterine duct and upper part of vagina.AMH may be detected in the serum from female embryo at 36 weeks of gestation to menopausal women;no AMH is secreted from primordial follicle;high expression of AMH can be detected in the granular cells of preantral follicles and antrum follicles; there is no expression of AMH in atretic follicle.AMH expressed in granular cells of growth follicles has the function to regulate the growth and development of follicles. It has the functions of inhibiting the initial recruitment of primordial follicles, reducing the response of preantral follicles and antrum follicles to the effect of FSH and influencing cycle recruitment.In recent years,the relation between AMH and PCOS has been paid more attention by researchers.Polycystic ovarian syndrome(PCOS) is one of the most common endocrine disorders.Its features include infertility,menstrual disorder, hirsutism and polycystic change secondary to bilateral ovary enlargement.PCOS, involving 5%-10%of Women of childbearing age all over the world,was discovered by Stein and Leventhal in1935,however,the pathophysiology of the condition is unclear. Its damage to the body and mental of women due to amenorrhea and infertility and the risk of frequent occurrence of endometrial cancer,breast cancer and cardiovascular and cerebrovascular diseases secondary to continuous stimulation from estrogen have attracted the more and more attention.Anovulation and rare ovulation are the major factors resulting in infertility in PCOS and patients of PCOS with repeating infertility after ovulation-promotion often need the therapy of assisted reproductive technology.Due to its high clinical diversification,difficulty in controlling the procedure of ovarian hyperstimulation and the quality concerns of ovum,it has become one difficult point and also a hotspot.Extensive studies have explored the relation between the expression of serum AMH and abnormal development of the follicles in PCOS patients and found that serum AMH level on the third day of menstrual period is associated with the number of antrum follicles with the diameter of 2-5mm.In patients of PCOS,their numbers of antrum follicles as well as their serum AMH are higher than health women by 2-3 times,and AMH is relevant to the mechanism of PCOS.However,there still is no report in China involving the effect of serum AMH on the clinical features of IVF therapy during ovarian hyperstimulation in IVF-ET therapy performed in patients of PCOS.With the continuous development of assisted reproductive technology(ART), In-vitro fertilization and Embryo transfer(IVF-ET) has became an important measure in treatment of sterility.And,one successful clinical pregnancy after IVF-ET is the result of cooperation of high-quality embryo,suitable endometrial receptivity and good endocrine level.The factors affecting the outcome of IVE-ET involve many aspects,including age,type of infertility,cause of infertility,basic serum follicle stimulating hormone(FSH),dosage of gonadotropic hormone(Gn),thickness of endometrium,serum estrogen(E₂) level,retrieved oocyte number and etc.In recent years,AMH has received quite attention in female-reproductive scope and the research on it is also becoming deepened.It has been found that AMH can be used to early and exactly predict the change in ovarian reserve.AMH is secreted by granular cells of preantral follicles and antrum follicles to inhibit the growth of follicles, prevent the premature consumption of follicles and to maintain ovarian reserve,thus, in clinical it has been used as a marker of ovarian reserve and to monitor the depletion of follicles resulted from ovary aging.AMH keeps a close relation to the status of ovary and the development of follicles and its relationship with the clinical features after IVF-ET treatment has not acquired a consistent conclusion.There are lots of disputations in the prediction of outcome of IVF therapy by AMH level.There are no reports in home involving the prediction of IVF pregnancy by AMH in serum or follicle fluid after controlled ovarian hyperstimulation(COH). For such an important role played by AMH in female reproduction,further study on it is necessary in clinical aspect,so as to provide practical support for the diagnosis and treatment of patients of PCOS and clinical instruction for IVF therapy.Partâ… The relationship between AMH and clinical characteristics during in vitro fertilization in PCOS patientsOBJECTIVEThrough comparative analysis of the relation of clinical characteristic and AMH level in serum and follicle fluid between PCOS and control group,preliminary explore the relation of AMH to the clinical features of PCOS treated by IVF-ET.METHODS1.Serum and follicular fluid were obtained from 64 women undergoing IVF/ICSI-ET in our unit.The patents were divided into two groups according to the Rotterdam criteria for diagnosis of PCOS.One group including 30 patients of PCOS and another as the control group(34 cycles).The long stimulation protocol was used for COH in all cycles.2.At 9 am on the day of oocyte retrieval,2ml of vein blood was collected and put into dry glass centrifuge tube prior to intravenous general anaesthesia;then kept the separated serum for later determination.Follicles fluid with the diameter between 15 and 18mm was retrieved by ultrasound-guided oocyte retrieval;left the first tube of clear mixed follicle fluid into a centrifuge tube and removed the granular cells; then kept it for later determination.3.Record the individual retrieved oocytes,number of occyte fertilization and number of total embryos.A positive pregnancy test was defined by＞50 mIU/mL of plasmaβ-hCG on day 14 after embryo transfer.An ultrasound scan 3 weeks after a positive pregnancy test confirmed a clinical pregnancy.4.The AMH levels in the follicular fluid and serum were assayed by a commercially available nzyme-linked immunosorbent assay(ELISA) kit.5.Statistical analysis of all data was performed with SPSS13.0.RESULTS1.Comparison of the clinical characteristics between the patients of both groupsPatient age,infertility period,BMI,basic serum FSH,LH,E₂ level and LH/FSH,days of Gn using,E₂,LH,P level on the day of HCG,thickness of endometrium on oocyte retrieval day and number of retrieved oocytes were no differences between PCOS and control group.(P＞0.05).The number of basic antral follicles was significantly higher in pcos group than that in control group(P＜0.05); The gonadotropin doses was significantly decreased in PCOS groups than that in control group(P＜0.05).Neither fertilization rate,cleavage rate,rate of high-quality embryo,implantation rate,pregnancy rate nor abortion rate and ongoing pregnancy rate were detected significant differences differences between PCOS group and control group.(P＞0.05).2.Comparison of AMH in serum and follicle fluidThe AMH level in serum and follicle fluid was significantly higher in PCOS group than that in control group(P＜0.05).CONCLUSIONS1.After performing IVF-ET ovarian hyperstimulation,we studied the relation between the AMH level in serum and follicle fluid on oocyte retrieval day and the clinical characteristic and outcomes in patients of PCOS group and found the increase of AMH level in serum and follicle fluid was related to the number of basic antral follicles.AMH might reflect the clinical characteristic in patients of PCOS.AMH level in patient of PCOS was higher than that in patients of control group and the application amount of Gn in PCOS group was lower than that in control group,AMH might reflect the reactivity of ovary to Gn,which might suggest a certain clinical significance in guiding clinical protocol and the application of Gn.2.The abortion rate was significantly higher in PCOS than that in control group,which suggested that higher AMH level in follicle fluid is a novel mechanism for abortion.Partâ…¡:The relationship between AMH and IVF-ET outcomeOBJECTIVEThrough the analysis of the AMH level in serum and follicle fluid at oocyte retrieval day after ovarian hyperstimulation,to study the prediction action of AMH on pregnancy after IVE-ET treatment and analyze its association with the factors relevant to pregnancy.METHODSThe material and method as partâ… .RESULTS1.AMH level in serum and follicle fluid of patient between pregnant and non-pregnant women in PCOS group.The patient's age,infertility duration,BMI,basic serum FSH level,days of Gn using,basic serum FSH,LH,E₂ level and LH/FSH,days of Gn using,E₂,LH,P level on the day of HCG,thickness of endometrium on oocyte retrieval day and number of retrieved oocytes were no differences between pregnant and non-pregnant women in PCOS group(P＞0.05).The rate of fertilization,cleavage rate and rate of high-quality embryo were no differences between the two groups(P＞0.05).AMH level in serum and follicle fluid was significantly higher in pregnant patients than that in non-pregnant patients(P＜0.05).2.AMH level in serum and follicle fluid of patient between pregnant and non-pregnant women in control group.The patient's age,infertility duration,BMI,basic serum FSH level,days of Gn using,basic serum FSH,LH,E₂ level and LH/FSH,days of Gn using,E₂,LH,P level on the day of HCG,thickness of endometrium on oocyte retrieval day and number of retrieved oocytes were no differences between pregnant and non-pregnant patient in control group.(P＞0.05).Rate of high-quality embryo was increased in pregnant patients compared with non-pregnant patients(P＜0.05).Neither the recovery rates of oocytes,fertilization rate,cleavage rate nor rate of high-quality embryo were detected significant differences differences between pregnant and non-pregnant patient.(P＞0.05).AMH level in serum and follicle fluid was significantly higher in pregnant patients than that in non-pregnant pregnant and non-pregnant patient.(P＜0.05).3.Correlations of AMH in serum and follicle fluid concentrations with pregnant during in vitro fertilization.As is shown by the graph,the AMH levels in follicular fluid of all pregnancy patients in POCS group are greater than 6.0 ng/ml,and the ratio of absence of pregnancy whose AMH levels in follicular fluid were lower than 6.0 ng/ml has reached 81.12%in all absence of pregnancy patients of PCOS group.But in control group the patients have to be pregnant whose AMH levels in follicular fluid are all greater than 2.0 ng/ml,and approximately 55.56%absence of pregnancy patients in this group have lower AMH levels in follicular fluid than 2.0 ng/ml.CONCLUSIONS1.The AMH level in serum and follicle fluid in pregnant PCOS patients was significantly higher than that in pregnant patients in control group,which showed AMH could predict the outcome of pregnancy after IVF.2.We showed that the AMH levels in serum and follicle fluid from women undergoing IVF whose embryo failure to implantation were statistically significantly lower than in the in serum and follicle fluid of those whose embryo became implantation.