Effects Of Exogenous Adiponectin Supplementation During Early Pregnant Mice With Polycystic Ovary Syndrome On Metabolic Syndrome Of Adult Female Offspring

Objectives Polycystic ovary syndrome(PCOS)is a heterogeneous gynecological endocrine disease with reproductive and metabolic abnormalities,which has a serious impact on women of childbearing age.Although the exact cause of PCOS is not clear,some clinical observations and animal studies have shown that PCOS has been confirmed to have a certain genetic background,and maternal hyperandrogenemia may be a contributing factor.Compared with healthy women,the vast majority of PCOS patients have hyperandrogenemia or hyperandrogenemia-like symptoms,and excessive androgen exposure during pregnancy may affect the development and metabolism of female offspring.In this experiment,we successfully established the model of PCOS mice,and the maternal PCOS mice were supplemented with exogenous adiponectin(APN)in the early stage of pregnancy.By tracking the endocrine indexes of female offspring after adulthood,we observed the main effects of APN intervened during their embryonic phase on metabolism syndrome or endocrine disturbance.At the same time,the relative expressions of m RNA and protein of phosphorylated AMPK,PI3 K,and Akt in the ovary tissue of adult female offspring were detected to further explore the mechanism of APN.Methods 1.The maternal PCOS mouse model was established by feeding fat diet combined with subcutaneous injection of dehydroepiandrosterone(dehydroepiandrosterone,DHEA).The success of PCOS model was verified by the trend of body weight,the level of serum testosterone and the results of HE staining in ovarian tissue.After successful modeling,living female mice were randomly divided into three groups: maternal-normal group(normal female mice were injected with normal saline in early pregnancy,n = 10),maternal-PCOS control group(PCOS model mice were injected with normal saline in early pregnancy,n = 20)and maternal-APN administration group(PCOS model mice were injected with APN in early pregnancy,n = 19).2.The maternal PCOS mice were supplemented with exogenous APN in early pregnancy,and thereafter the concentrations of serum APN and testosterone in part pregnant mice were randomly detected to verify whether there was hypoadiponectin in PCOS pregnant mice and conform the dose of adiponectin supplement.The reproductive ability of maternal mice was evaluated by observing the pregnancy rate and average litter size of remaining pregnant mice.3.The female offspring of each group formed three offspring-groups: offspring-normal group(n = 6),offspring-PCOS control group(n = 6)and offspring-APN administration group(n = 6).The growth,development and endocrine indexes of female offspring were monitored,and then the effects of adiponectin on metabolism syndrome or endocrine disturbance were analyzed.4.The relative expressions of phosphorylated AMPK,PI3 K,Akt m RNA and protein in the ovary of offspring mice were detected by q RT-PCR and Western-Blot methods to investigate the effect of exogenous adiponectin supplemented during early pregnancy of maternal PCOS mice on expressions of phosphorylated AMPK,PI3 K,and Akt in ovary tissues of adult female offspring mice.5.The experimental data were expressed by Mean ±SD(x ±s).Independent-Samples t Test or one-way ANOVA analysis was performed by SPSS25.0 statistical software.Results 1.PCOS mouse model and the difference of maternal pregnancy outcome The body weight and serum testosterone level of mice in PCOS model group were significantly higher than those in normal control group(P < 0.01).The morphological examination of ovary in PCOS model group showed obvious polycystic changes in ovary,mainly characterized by thinning of granulosa cell layer,decrease of corpus luteum and increase in the number of atretic follicles,indicating that the model of PCOS mice was established successfully.Through observation and comparative analysis,the average litter size of maternal PCOS mice(maternal-PCOS control group)was significantly lower than that of maternal normal mice(maternal-normal group),while the pregnancy rate(P<0.01)and average litter size(P<0.05)of maternal PCOS mice of APN administration during early pregnancy(maternal-APN administration group)were significantly higher than those of maternal-PCOS control group.It was suggested that APN intervention in early pregnancy could reduce the loss of PCOS mice embryos to some extent and improve the breeding ability.2.Adiponectin supplementation led to the normalization of PCOS-like endocrine phenotype in offspring induced by androgen exposure during the embryonic period.The serum testosterone level of maternal-PCOS control group(1.47±0.38ng/ml)was significantly higher than that of maternal-normal group(0.14±0.05 ng/ml),while the serum testosterone level of maternal-APN administration group was significantly lower than that of maternal-PCOS control group(P<0.01),and significantly higher than that of maternalnormal group(P<0.05).It is suggested that APN intervention in early pregnancy can reduce hyperandrogen exposure during the embryonic period to some extent.The serum testosterone level of the female adult offspring of the PCOS mice(offspring-PCOS control group)(0.23±0.10 ng/ml)was significantly higher than that of the female adult offspring of the normal mice(offspring-normal group)(0.11±0.09 ng/ml),and the time of interestrus characterized by the aggregation of a large number of granulocytes in the offspringPCOS control group(10.0±1.0d)was significantly longer than that of the offspring-normal group(4.0±0.0d).Compared with the offspring-normal group,there were noticeable PCOSlike reproductive endocrine disorders centered on hyperandrogenemia and insulin resistance in the offspring-PCOS control group.However,the serum testosterone level of female adult offspring of PCOS treated with APN during their embryonic period(offspring-APN administration group)(0.08±0.05 ng/ml)was significantly lower than that of offspring-PCOS control group(P<0.05),and lower than that of offspring-normal group,but had no significant difference(P > 0.05).The phenotypes of metabolic disorders such as obesity,insulin resistance,impaired glucose tolerance and hyperlipidemia were significantly improved in offspring-APN administration group(P<0.05).3.Adiponectin supplementation in early pregnancy of PCOS maternal mice affected the expression of phosphorylated AMPK,PI3 K and Akt in the ovary of offspring. Compared with the offspring-normal group,the expression levels of p-AMPK,p-PI3 K and pAkt in the ovary of the offspring-PCOS control group were significantly decreased(P<0.05),while the expressions of p-AMPK,p-PI3 K and p-Akt were significantly increased in the offspring-APN administration group(P<0.05).Conclusions After treatment with adiponectin in early pregnancy,the success rate of pregnancy in maternal PCOS mice was significantly improved,and the adverse effects of maternal high androgen levels during pregnancy on female offspring were effectively reduced,and the PCOS-like endocrine phenotype and various metabolic disorders of PCOS in offspring adult mice were corrected.The AMPK/PI3K-Akt pathway in the ovary of offspring of PCOS mice was inhibited,while it would be markedly activated when the maternal PCOS mice received APN treatment during early pregnancy.These suggested that APN might improve the disorder of endocrine metabolism in adult female offspring of PCOS mice through AMPK/PI3K-Akt signal pathway.