| Research background:Infertility accounts for about 10%to 12%of the population of childbearing age,and 30%of the female factor is caused by ovulation disorders.Egg development is carried out in the form of follicles in the ovaries of women.During the development process,it is regulated by hormones and growth factors.When abnormal maturation occurs,different forms of ovulation disorders occur.The TGF-? family is an important regulator in oocyte maturation.Studies have found that egg maturation is closely related to the surrounding granular cells.Both GDF-9 and AMH belong to the TGF-P family and not only play an important role in oocyte maturation,but also have important indicators in the treatment and diagnosis of infertility.In this paper,the clinical application of GDF9 and AMH in patients with ovulation disorders was evaluated and analyzed,and the application of GDF9 in vitro culture of mammalian oocytes was explored.By analyzing the transcription group of ovulation disorders,the role of GDF9 and AMH in follicular growth and development was further revealedPart I:GDF-9(Growth Differentiation Factor 9)Applied to the Evaluation of Ovulation DisordersObjective:1 A quantitative analysis of the expression of granulocyte GDF9 in women with ICSI was carried out in this hospital,and the sensitivity of the patient to hyper-emission hormone(mainly FSH)was analyzed in order to find out the correlation between the two.The effects of GDF9 and FSH on the maturation and embryonic development of mouse oocytes were observed in IVM of mice.Methods:Selection From May 2017 to January 2018 in our line of follicle intracytoplasmic sperm injection(intracytoplasmic sperm injection,ICSI)of 72 cases of female patients,according to obtain the average diameter>when you pick up the egg number of 14 mm follicles grouping:follicular number 5 or less for low ovarian response group,6?19 for reaction in the group,20 as high or higher reaction group,24 cases each.The expression of GDF-9 in granulocytes of different reaction groups of ovaries and the correlation between GDF-9 expression and ovulation disorders were observed.Results:1.Among the 72 cases of IVF patients,24 were low in the ovaries,24 in the middle reaction group and 24 in the high response group.There was no significant difference in age,infertile years,basal FSH,E2 level and Gn dosage in each group(P>0.05).2.GDF-9 m RNA expression was found in the cells of the follicular wall granules and the granule cells of the patients,and the differences in the 3 groups were statistically significant(P<0.05).The relative expression of GDF-9 mRNA in the cells of follicular wall granule cells and oocytes in the low reaction group was lower than that in the medium reaction group and the high response group(P<0.05).3.The number of eggs in the low,medium and high reaction groups of the ovaries was:(3.45±0.97),(12.38±3.36),(24.38 + 4.86).The expression of GDF-9 m RNA was positively correlated with ovulation.Conclusion:GDF-9 is an indispensable growth factor in the development of follicles.This study found that the decrease of m RNA expression level was related to the low response of IVF super ovulation cycle.For patients with low super ovulation reaction,can try to apply restructuring GDF-9 stimulate primary follicular growth,increased sensitivity to designed.The Gn sinus follicle before,with r FSH regulation of follicular,improves ovarian function.It can reduce the dosage of Gn,make ovulation scheme more reasonable,and provide the possibility of using oocyte for infertility patients.Part ?:The Relationship between the Anti-Mullerian Hormone(AMH)and Ovulatory Disorders and the Outcome of IVF PregnancyObjective:Observe and explore the relationship between the antiovulatory hormone(AMH)and ovulation disorder,and the correlation with the pregnancy outcome of IVF-ET.Methods:Enzyme-linked immunosorbent assay(ELISA)to detect serum anti mullerian hormone levels,estradiol(E2),follicle stimulating hormone(FSH),luteinizing hormone(LH),progesterone(P)and testosterone(T)in 106 infertile women(observation group)and 102 women pregnant before the physical examination(control group)compares the data of the statistical analysis.106 infertile women(observation group)according to the classification of infertility factor,divided into immune factors group(n = 8),tubal factor group(n = 54),endocrine factors(n = 32),uterine factor group(n = 12).In addition,50 patients with primary IVF treatment were collected from December 2016 to December 2017.Results:The hormone levels of AMH and LH in the observation group were higher than that in the control group(P<0.05).The FSH level of the observation group was significantly lower than that in the control group(P<0.05).Pearson correlation analysis showed positive correlation between serum AMH and LH(P<0.05)and T(P<0.05).The serum AMH level in the observation group was negatively correlated with FSH(P<0.05).Serum AMH had no significant relationship with E2 level.There was a statistical difference in the AMH between the low ovulation array,the normal ovulation array and the high ovulation array in IVF.IVF in low for low egg array for the number of eggs FSH values,the usage of designed.The Gn is significantly higher than the normal egg arrays,high egg,embryo several array and clinical pregnancy rate is lower than the normal eggs,the eggs high array(P<0.01).Conclusion:AMH was positively correlated with ovulation number,AFC,ovary volume,embryo number and clinical pregnancy rate.FSH was negatively correlated with D2 days;there was no correlation with PSV,RI,sterility antibody.This study suggests that serum AMH level is related to follicular dysplasia,which is an effective measurement method for ovulatory dysfunction of sterile female.Part III Analysis on transcriptional expression of GDF9 and AMH genes of granulosa cells in abnormal fertilized eggs in IVFObjective:The oocyte maturity obtained by superovulation in patients with ovulation inhibition is not uniform.Therefore,the granulosa cells of normal fertilized eggs after IVF and the granulosa cells of abnormal fertilized eggs are collected for transcriptome sequencing analysis to find out the difference in expression and association of related genes between GDF9 and AMH.Methods:The cumulus cells of the oocytes were collected and stored in numbers.The fertilized eggs were observed after 16 hours of in vitro fertilization,and the cumulus cell samples of 2PN and ? 3PN were sent to Novogene Corporation for transcriptome sequencing.Results:The data is verified to be reliable this time.,The granulosa cell transcriptome data of the normal fertilized eggs after filtration is13.18G,and that of the abnormal fertilized eggs is 10.5G.Compared with abnormal fertilized oocytes,693 genes are up-regulated and 857 genes are down-regulated in normal fertilized oocytes.GO enrichment analysis show that the granulosa cell transcription level of abnormal fertilized eggs is significantly reduced,while the cells are in a stringent state,and the expression of AMH is significantly down-regulated.After the enrichment analysis of KEGG,it is found that the biological role of TGF-? signaling pathway in granulosa cells of abnormal fertilized eggs is inhibited,which ultimately leads to the polyspermy. |
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