The Outcomes Of IVF/ICSI In PCOS Patients And The Related Mechanism Of Its Influence On Neonatal Birth Weight

Polycystic ovary syndrome(PCOS)is a common reproductive endocrine disease characterized by chronic anovulation,hyperandrogenemia and insulin resistance,the prevalence of PCOS is estimated to be nearly 5% ～ 10% among reproductive-age women,approximately 50% ～ 70% of these women with infertility.When drug-induced ovulation or surgery fails for infertile women with PCOS,in vitro fertilization based assisted reproductive technology(ART)has become a treatment option for such women.However,studies have found that the rate of miscarriage and premature delivery,complications during pregnancy and infant hospitalization are increased after women with PCOS have conducted ART assisted reproduction.Other studies have shown that women with PCOS have increased pregnancy complications such as gestational diabetes mellitus,preeclampsia,etc.and the offspring are prone to birth weight changes.These adverse obstetric outcomes are significantly correlated with increased serum testosterone level of patients with PCOS.For PCOS patients undergoing ART assisted pregnancy,what are the effects of abnormal endocrine and metabolic environment on adverse pregnancy outcomes?What are the mechanisms involved in adverse pregnancy outcomes caused by intrauterine excessive androgen exposure? Thus,clinical data analysis was carried out on IVF/ICSI pregnancy outcomes of patients with PCOS and then a mouse excessive androgen model during pregnancy with PCOS was established to analyze the influence of high excessive androgen exposure on placenta and offspring.The study aim was to explore the influence of high excessive androgen environment on intrauterine growth and development of offspring and its relevant mechanism,to provide the theoretical guidance for clinically treating the patients with PCOS with assisted reproduction.Part Ⅰ: Assisted reproduction outcome of patients with PCOS undergoing IVF/ICSI and Influence on birth weight of newborns.Objective: Explore the pregnancy outcome condition of patients with PCOS after undergoing IVF/ICSI assisted reproduction and influence on birth weight of newborns after delivery.Methods: Adopt the women with PCOS undergoing IVF/ICSI assisted reproduction in Reproductive Medicine Center of our hospital from January 1 to December 31,2017 as the study object;adopt the women with simple tubal factor undergoing IVF/ICSI assisted reproduction over the same period as the control group;record the assisted reproduction information,pregnancy outcome and offspring birth condition of two groups of patients;compare the clinical pregnancy rate,biochemical pregnancy rate,ectopic pregnancy rate,abortion rate,premature birth rate,live birth rate,sex of newborns and birth weight of newborns of two groups of patients.Results:(1)Study 707 patients with simple tubal factor infertility,115 patients with PCOS.Analyze the pregnancy outcome of two groups,the results show that late abortion rate of patients of PCOS group is 4.11%,late abortion rate of patients of the control group is2.12%;compared with the control group,the difference has statistical significance(P<0.05);biochemical pregnancy rate,clinical pregnancy rate,ectopic pregnancy rate,early abortion rate,premature birth rate and stillbirth rate has no statistical difference between the two groups(P>0.05).After adjusting the confounding factors,conduct the multiple regression analysis on the pregnancy outcome of patients in fresh embryo transfer cycle and frozen thawed embryo transfer cycle respectively,the results show that: in fresh embryo transfer cycle,for biochemical pregnancy rate,clinical pregnancy rate,early abortion rate and live birth rate,the difference has no statistical difference between the two groups(P>0.05);in frozen thawed embryo transfer cycle,premature birth of patients with PCOS is increased 182%[OR(95%CI): 2.82(1.31,6.08),P=0.0082] compared with the patients with tubal factor,the difference has statistical significance(P < 0.05);for biochemical pregnancy rate,ectopic pregnancy rate,early abortion rate,late abortion rate,live birth rate and stillbirth rate,the difference has no statistical difference between the two groups(P>0.05).(2)Study the patients who bring into singleton and full term delivery,281 patients with simple tubal factor infertility,49 patients with PCOS.Analyze the birth weight of newborns of two groups of patients,the results show that birth weight(g)of newborns of two groups of patients has no statistical difference(control group:3318.42 ± 509.41 vs PCOS group: 3238.57 ± 569.20,P>0.05),further adjust the confounding factors,conduct the multiple regression analysis on birth offspring weight of singleton pregnancy patients in fresh embryo transfer cycle and frozen thawed embryo transfer cycle respectively,the results show that in Fresh embryo transfer cycle,birth weight of newborns of patients with PCOS is reduced 16.66 g [OR(95%CI):-16.66(-228.70,195.38),P= 0.8779] compared with the patients with simple tubal factor,the difference has no statistical significance(P>0.05);in frozen thawed embryo transfer cycle,birth weight of newborns of patients with PCOS is reduced 16.66 g [OR(95%CI):-16.66(-228.70,195.38),P= 0.8779] compared with the patients with simple tubal factor,the difference has no statistical significance(P>0.05).Conclusion: Late abortion rate of patients with PCOS is increased,after adjusting the confounding,it has been found that premature birth rate of patients with PCOS in frozen thawed embryo transfer cycle is significantly increased;therefore,it is suggested that the patients with PCOS should strengthen the management during pregnancy(particularly in mid-pregnancy period),increase the times of prenatal examination if necessary.Although the birth weight of newborns of patients with PCOS has tended downwards compared with the patients with simple tubal factor,the difference has no statistical significance and is worthy of further study.Part Ⅱ: Influence of PCOS excessive androgen exposure on fetal placenta development of mice.Objective: Build PCOS model of mice,simulate PCOS excessive androgen environment,explore the influence of excessive androgen on pregnancy outcome and birth weight of fetal mice.Methods: Select 6-8SPF level ICR mice,from 6.5 days after natural conception,inject subcutaneously DHEA oil agent(60 mg/kg·d)into the neck every day,simulate PCOS excessive androgen environment,build PCOS model group,and build the control group of simple corn oil injection;the mice should be continuously completed 12.5 days intervention and should be broken the neck to death in 18.5 days;collect the fetal mice and placenta specimens.(1)Compare the implantation rate,survival rate and average litter size of two groups of fetal mice;(2)Compare the weight,placental weight and placental efficiency of two groups of fetal mice.(3)The morphological changes of placental tissue of mice using hematoxylin-eosin staining method.Results:(1)Influence of PCOS excessive androgen exposure on pregnancy outcome of fetal mice: average litter size of PCOS model group is 12.14 ± 1.95(pcs),survival rate is77.27%;average litter size of the control group is 16.5 ± 1.64(pcs),survival rate is91.67%;average litter size and survival rate of PCOS model group is lower than the control group,the difference has statistical significance(P<0.05).(2)Influence of PCOS excessive androgen exposure on placenta development of mice: weight of fetal mice of PCOS model group is 1324.31 ± 183.67(mg),placental weight is 93.55 ± 16.08(mg);weight of fetal mice of the control group is 1477.75 ± 176.47(mg),placental weight is102.93 ± 17.82(mg);weight of fetal mice,placental weight of PCOS model group is significantly lower than the control group(P<0.05);placental efficiency of PCOS model group is 14.34 ± 1.98,placental efficiency of the control group is 14.65 ± 2.36,the difference between two groups has no statistical significance(P>0.05).(3)Influence of PCOS excessive androgen exposure on morphology of placental tissue of mice: After placental HE staining,it has been found that ratio of area of placental spongiotro-phoblast layer to total placental area,proportion of area of spongiotro-phoblast layer of PCOS model group is 0.254 ± 0.439,proportion of average area of the control group is 0.35 ±0.024,PCOS model group is lower than the control group,the difference has statistical significance(P<0.05).Conclusion: In PCOS mice model,excessive androgen environment may increase the abortion rate of mice and reduce the survival rate;weight of fetal mice and placenta is significantly reduced,proportion of area of placental spongiotro-phoblast layer is declined.The results show that PCOS excessive androgen exposure may affect the development and function of placenta,and then affect the growth and development of fetus.Part Ⅲ: Influence of PCOS excessive androgen exposure on placental function of mice and preliminary discussion on mechanism.Objective: Explore the mechanism research on influence of excessive androgen exposure on change of placental structure and function in PCOS mice model.Methods: Mark CD31 and determine the placental vessel density using immunohistochemistry;detect mRNA expression level of pro-angiogenic related genes,glucose transporter and amino acids transporter using real-time quantitative PCR method.Results:(1)After detecting the vessel density of mice placenta,it has been found that mean vessel density of PCOS model group is 143.83 ± 21.85,mean vessel density of the control group is 123.73 ± 12.64;vessel density of PCOS model group is higher than the control group,the difference has statistical significance(P<0.05).(2)After detecting mRNA expression level of pro-angiogenic related genes of two groups of placenta,it has been found that expression level of Vegf,Kdr,Flt1,Ang,Mmp2,Pdgfa,Pdgfb,Plgf2 of PCOS model group is significantly increased(P<0.05).(3)Detect mRNA expression of glucose transporter of two groups of placenta,it prompts that expression level of Slc2a1,Slc2a2,Slc2a3,Slc2a9 and Slc2a12 of PCOS model group is significantly reduced(P<0.05);mRNA expression level of amino acids transporter of two groups of placenta,expression level of Slc38a1,Slc38a2,Slc38a4,Slc43a2,Slc7a5 and Slc7a8 of PCOS model group is significantly reduced(P<0.05).Conclusion: Mean vessel density of placental tissue in PCOS model group is significantly increased,expression level of vascular-related genes is increased,it prompts that excessive androgen exposure affects placental angiogenesis,may be inducing factor of causing placental vascular induced diseases(such as preeclampsia).In addition,mRNA expression level of glucose transporter and mRNA expression level of amino acids transporter of fetal mice in PCOS model group is significantly reduced,it prompts that excessive androgen exposure will affect the transport capacity of placental nutrients,and then lead to birth weight loss of fetal mice.