Effects Of Bisphenol A On Reproductive Capacity Of SD Rats And Analysis Of MiRNA Differential Expression

Objective: Bisphenol A is a class of environmental endocrine disruptors(EDCs)that are ubiquitous in the external environment,with wake estrogen activity,exposure to Bisphenol A will have a wide range of adverse effects on human health,can target reproductive organs and damage women’s reproductive ability,but the mechanism of occurrence is not yet clear.In this study,the reproductive ability of offspring was assessed by observing the fertility of the first and second generations,uterine and ovarian coefficients,ovarian histopathology and other indicators by establishing a SD rat infection model,the intergenerational effect of different doses of Bisphenol A on female reproductive function,and the transcriptometry sequence of the control group and the highest dose group of F1 generation(F1PND56),and the database was used to predict the differential miRNA target gene.G0 function and KEGG pathway analysis to explore miRNA-target gene-phenotype regulation.Materials and methods: A cross-generational model of female rat reproduction was constructed by continuous gastric infection Bisphenol A,and different doses of Bisphenol A infection were observed on female mouse fertility,GD20 fetal mice in general,PND21 and PND56 body weight,uterine coefficient,and ovarian coefficient.Follicle counts are performed on offspring ovarian tissue and miRNA sequencing and bioinformatic analysis of F1 generation Bisphenol A high-dose group versus control group ovarian tissue.Results:(1)Fertility and early embryonic development of SD ratsUnder Bisphenol A exposure,the average number of implantations and uterine fetal weight of F0 and F1 surrogate mice showed a downward trend,and the rate of pre-implantation loss showed an increasing trend,with F0 generation being significant at 25 ug/kg(p<0.05),F1 generation2.5 ug/kg significant(p<0.05),F1 generation 25000 ug/kg pregnancy rat absorption fetal rate significantly increasing(p<0.05),but the cage time,fertility index,gestational rat mortality and mating rate,average corpus luteum required for successful mating of two generations of pregnant mice,Stillbirth rates were not significantly affected.(2)Bisphenol A exposure to offspring female rats in generalBisphenol A intrauterine exposure will significantly increase the weight of female fetal mice dissected by F0 and F1 generation GD20(P<0.05);the uterine coefficient of female mice in the F1 generation,F2 generation de-lactation stage(F1,F2PND21)and F1 generation sexual maturity stage(F1PND56)will be significantly reduced(p<0.05),but the ovarian coefficient will decrease in the F1 generation delactal stage(F1PND21),the F1 generation sexual maturity stage(F1PND56),and the F2 generation sexual maturity stage(F2PND56)will decrease.(3)Follicle count and ovarian morphological changesThe total number of follicles of 25 ug/kg and 250 ug/kg in the F1 generation delacting stage(F1PND21)increased,mainly due to the increase of locked follicles,and the follicle morphology of the F1 generation sexual maturity stage(F1PND56)was morphologically atrophied compared with the F1 generation de-lactation stage(F1PND21),the granulocyte arrangement was disordered,the gap widened,and the follicle interstitium increased.The number of morphological forms of ovarian follicles at all levels decreased at the F2 generation sexual maturity stage(F2PND56),and cystic follicles were seen,and the lock-ate follicles of 250ug/kg and 2500ug/kg increased significantly.(4)Effect of differential expression of miRNAComparing the high-throughput sequencing of miRNAs in the A and F1PND56 groups of F1PND21 and F1PND56,16 miRNAs were upregulated and 14 miRNAs were down-regulated in the PND21 F group compared with the PND21 A group;99 miRNAs were upregulated and103 miRNAs were down-regulated in the PND56 F group compared with the PND21 F group;and 25 miRNAs were upregulated and 27 miRNAs were upregulated in the PND56 F group compared with the PND56 A group.The target gene was predicted to be LCP2 and JTB,and the GO function analysis of differential miRNAs mainly regulates cellular metabolic processes,performs biological regulation,and is significantly enriched in phosphorylation metabolism of cell metabolism.KEGG metabolic pathways are significantly enriched in the categories of signaling and endocrine systems.conclusion:(1)Sustained exposure to Bisphenol A during pregnancy may increase the weight of the offspring fetal mice,reduce the ovarian coefficient and uterine coefficient,thereby causing damage to the female gonads;high-dose Bisphenol A-treated animals have fewer ovarian mature follicles and corpus luteum,and the atresia follicles are increased,and cystic follicles are visible.(2)Continuous exposure to Bisphenol A can lead to changes in miRNA expression in offspring ovarian tissue,with varying degrees of changes during lactation and sexual maturity at different exposure concentrations.(3)DEG enrichment analysis showed that it can regulate the cellular metabolic process,phosphorylated metabolic pathway affects ovarian function,KEGG metabolic pathway is significantly enriched in signaling,endocrine categories,and may affect Lcp2 expression,may cause premature aging of the ovaries through immune response,but the specific regulatory mechanism still needs to be further verified.