Perinatal Combinational Exposure To Bisphenola And A High-fat Diet Contributes To Intergenerational Dysregulation Of Cardiovascular And Metabolic Systems In Mice

Background:There has been a worldwide increase in the prevalence of cardiovascular and metabolic diseases,which have made them a grave public health concern.In addition to genetic susceptibility,other risk factors for hypertension and metabolic diseases include lifestyle and environmental factors,especially nutrition.Furthermore,exposure of rodents and humans(F0)to some adverse environmental factors can cause phenotype changes and increased susceptibility to diseases,with these adverse outcomes being possibly passed down to future generations(F1,F2,etc.)even without direct exposure.This is considered the intergenerational inheritance of the adverse effects of environmental factors.Bisphenol A(BPA)is a typical endocrine-disrupting chemicals(EDCs)with estrogenic activity.It is the precursor of polycarbonate and epoxy resin and is widely used in food packaging,medical devices,and other daily products.Exposure to BPA or a high-fat diet(HFD)during pregnancy has been shown to increases the risk of hypertension and obesity in offspring.However,there are few studies on the effects of combined exposure of the BPA and HFD in the perinatal period.In addition,high energy diets are common in modern society,it is necessary to determine the health effects of combined exposure to high energy diets and EDCs.Objective : Both BPA and HFD exert unfavorable effects on cardiovascular and metabolic regulation of homeostasis.BPA and HFD often coexist in modern lifestyles;however,the long-term effects of simultaneous exposure of mothers to BPA and HFD during the perinatal period on the cardiovascular and metabolic systems of the offspring remain unclear.Therefore,the purpose of this study was to explore the effects of perinatal HFD combined with BPA exposure on the cardiovascular and metabolic systems of the first generation and even the second generation,and to explore the relationship between maternal multiple adverse factors exposure to metabolic and cardiovascular regulation disorders in offspring.To provide a scientific basis for exploring the impact of perinatal exposure to multiple adverse factors on intergenerational health.Methods:We established a female ICR mice model that was exposed to BPA,HFD,and HFD+BPA in the perinatal period,the F0 generation mice were divided into 4 groups: a control low-fat diet(LFD)group;BPA group(500μg/kg/day);HFD group;HFD+ BPA group(500μg/kg/day BPA combined with HFD).After treatment for 10 weeks,the female mice were mated with age-matched healthy males to produce F1 offspring.All F1 mice received an LFD.The F1 mice were mated with untreated 12-weekold males/females to produce F2 offspring.A vaginal plug indicated successful conception;subsequently,the pregnant mice were housed individually throughout the gestation period until postnatal day 21.The F2 mice received a HFD again at the age of 13 weeks to determine amonggroup differences in metabolic and cardiovascular phenotypes.F2 mice received an LFD for 12 weeks.Subsequently,some mice received a HFD from the 13 th week to the 21 st week,while the remaining mice received an LFD until the 21 st week before being sacrificed.During the period,the body weight and glucose tolerance of all mice were observed and recorded,the blood pressure of each generation of mice with 4 different intervention methods was detected by the tail-cuff method,the cardiac structure of each generation of mice was detected by cardiac ultrasound,and wheat germ agglutinin fluorescent staining(Wheat germ agglutinin)was used.Agglutinin staining,WGA)was used to analyze the size of cardiomyocytes in F2 generation mice,metabolic cages were used to detect the difference in energy metabolism in mice,hematoxylin-eosin staining(HE)was used to analyze the morphology of F2 generation brown fat,and real-time fluorescent quantitative polymerase Chain reaction(q PCR)to detect the transcriptional expression level of key regulatory molecules,enzyme linked immunosorbent assay(ELISA)to determine the content of related hormones in serum,Western blot(WB)to determine the protein expression levels in tissues,and the possible mechanisms were explored by bisulfite sequencing to detect DNA methylation of key molecules.Results:(1)For the F0 generation,perinatal exposure in the HFD group and the HFD+BPA group significantly increased body weight and impaired glucose tolerance compared with the LFD group in metabolism,but there was no significant difference between the HFD group and the HFD+BPA group.Systolic blood pressure was significantly higher in perinatal BPA and HFD+BPA exposure compared with LFD group,but there was no significant difference between BPA group and HFD+BPA group.(2)For male mice of F1 generation,the body weight of F1 generation male mice from the F0 generation BPA group,HFD group and HFD+BPA group was significantly higher than the offspring of the F0 generation LFD group,but glucose tolerance and blood pressure were significantly increased.Only the F1 generation male mice from the F0 generation BPA+HFD group were significantly impaired glucose tolerance and significantly increased in blood pressure.For female mice of F1 generation,the body weight of female F1 generation mice from the F0 generation BPA group was significantly higher than the offspring of the F0 generation LFD group,and the F1 generation female mice form F0 generation BPA group and HFD+BPA group were no significant difference in the body weight,but the glucose tolerance of the F1 generation female mice in the F0 generation HFD+BPA group was significantly impaired compared with the F1 generation female mice in the F0 generation LFD group.(3)For the F2 generation mice,compared with the maternally derived F2 group from the F0 generation LFD group,the HFD+BPA group in the maternally derived F2 group showed a significant increase in systolic blood pressure,an increase in the area of cardiomyocytes,and changes in cardiac structure,as well as aortic thickness,decreased e NOS expression and increases in serum insulin.However,no changes in cardiovascular-related phenotypes were observed in the paternally-derived F2 mice.In terms of metabolism-related phenotypes,neither body weight nor glucose tolerance were significantly changed in F2 mice under the LFD diet.(4)When the F2 generation mice were given HFD intervention again,the paternally derived F2 generation female mice from the F0 generation HFD+BPA group showed significant changes in the metabolic phenotype compared with the paternally derived F2 group from the F0 generation LFD group,which was manifested in more weight gain,more significant impaired glucose tolerance,brown fat tissue whitening and reduced energy expenditure,but no significant changes in energy metabolism-related phenotypes in paternally-derived F2 male mice.Conclusion: Perinatal HFD combined with BPA exposure will not only increase itself blood pressure,but also cause increased blood pressure,cardiac structural remodeling,increased cardiomyocyte area,insulin resistance,decreased energy expenditure,and increased obesity susceptibility to at least the next two generations.The metabolic phenotypes were mainly exhibited in paternally derived mice,while the cardiovascular related phenotypes were mainly exhibited in maternally derived mice.Cardiovascular phenotype effects were sex-specific.Epigenetic changes may be the underlying mechanism of perinatal high-fat diet combined with bisphenol A exposure contributes to the disturbance of metabolism and cardiovascular regulation in offspring.