Effects Of Di(2-ethylhexyl)phthalate Exposure During Pregnancy On The Placenta Tissue Structure And Related Mechanism

Background Di-(2-ethylhexyl Phthalate(DEHP)is two phthalic acid lipid is the most commonly used,at room temperature for liquid oily compound,the appearance of a colorless,low melting point,high boiling point,poor water solubility,easily soluble in a variety of organic solvents,because of its strong softening effect,and high safety,so the same is widely used in the process of adding all kinds of plastic toys,food,medicine,plastic utensils,wallpaper,cleaning agents,lubricants and other personal care products.When the DEHP in the environment increases,the accumulation of DEHP in the body,when the concentration of DEHP in the blood reaches a certain concentration,easy to lead to acute poisoning.In recent years,with the industrial pollution,the various toxic effects of DEHP on human body gradually appear,and the research on human health of DEHP is also the focus of the current.The results suggest that the harm of DEHP to human health is mainly manifested in acute toxicity and toxicity to various organs.Among them,the impact on pregnancy and fetus is the current research hotspot.With regard to the pathogenesis of DEHP,current studies have demonstrated that it plays an important role not only in the functional and structural level of the organ level,but also in the functional and structural level of the protein molecule.However,the specific pathogenesis remains unclear,and more randomized controlled trials are needed to further confirm the data.Its impact on the reproductive system has attracted the attention of the majority of scientific research workers.Pregnant women and children are more sensitive to DEHP exposure than the general population.DEHP exposure is associated with diseases such as uterine fibroids,and may cause premature birth and delayed growth and development of offspring.Adverse effects of DEHP exposure and potential reproductive health in pregnant women should be further studied.Objective To study the different dose of DEHP exposure on the growth and development of fetal rats,study the effect of expression of fetal rat gene from m RNA and protein level of different doses of DEHP exposure to the placental tissue structure and its research mechanism,effects of pregnant rats and fetal growth and development,exploration the reproductive effects caused by DEHP,thus,provide a reference for further clear the damage of DEHP.Methods 60 pregnant Wistar rats were divided into 4 groups randomly,15 rats in each group,the control group only received 15 corn oil,group DEHP were randomly divided into three groups,respectively,the low dose group(100 mg/kg),middle dose group(500 mg/kg),high dose group(1000 mg/kg),on the 12 day of pregnancy every day given DEHP by gavage for nineteenth days of pregnancy,pregnant mice were killed.To observe the general situation of DEHP on pregnant mice and its influence on pregnancy outcome.Placental quality,the number of live births,monstrum number,fetal body,body length and tail length and each organ weight,the differences between the groups in these indicators;the expression of real time RT-RCR and Western Blot method by a further test method to detect the AKr7a3 gene,ugt2b37 gene,GSTA5 gene,Sult2a1 gene,Fmo1 gene,Maob gene,Aldh1a3 gene and cyp2f4 gene the differences between the various.Result 1 pregnant rats were generally in good condition,Mao Zhengqi,gloss,feeding and drinking were normal before the start of each group;the weight of rats no significant difference(P>0.05);during the experiment,the body weight of rats increased significantly,but the experimental group increased significantly less than the control group,there were statistically significant differences between the three groups(P<0.05);the weight difference,with the increase of DEHP dose,weight gain decreased,the difference was statistically significant(P<0.05).2 Normal control group rat placenta is presented for central thick edge thin,the color is dark red,mainly by the maternal fetal surface and surface composition,with amniotic membrane covered in fetal surface,amniotic membrane color version of the transparent,smooth surface;the middle dose group of placental dark volume decrease light weight,thin,central.In the high dose group,the part of placenta was congested and edematous,the volume increased,the weight increased,and the amniotic membrane was yellow and cloudy.HE staining showed that the normal control group the level of placental structure clear,cell morphology rules,trophoblast cells arranged;morphology of low dose group of placental cells was normal,no obvious change in DEHP dose group;placental trophoblast sponge thinning,trophoblast cell shrinkage,lost with polycythemia;high dose group of DEHP placenta of each layer boundary is not clear.Sponge trophoblastic layer became thin,the cytoplasm of space network.3 There was no significant difference between the experimental group and the control group and the experimental group in the number of group litters and the sex ratio of the fetus(P>0.05).There were no statistically significant differences in the number of live litter and the number of fetuses per group between the experimental group and the control group(P>0.05).The number of live fetuses in the middle dose group and the high dose group was significantly lower than that in the control group,the difference was statistically significant(P<0.05).Fetal body weight,body length and tail length in the experimental group was significantly lower than the control group,the difference was statistically significant(P<0.05);comparison between groups,between different dose groups of above indexes have difference,and with the increase of DEHP,these differences are more obvious,the difference was statistically significant(P<0.05).Compared with the control group,the experimental group was significantly smaller than the control group(P<0.05).4 Detected by gene chip data analysis showed that the AKr7a3,ugt2b37,GSTA5,Sult2a1,Fmo1,Maob,Aldh1a3,and Cyp2f4 gene expression level and DEHP concentration in a dose-dependent manner,may be the key gene in placenta of pregnant rats with different doses of DEHP after exposure to influence the expression level of.5 Fluorescent quantitative PCR showed that the gene expression in each group was compared,and the expression of Sult2a1 gene and Maob gene increased with the increase of DEHP concentration.The difference between the two groups was statistically significant(P<0.05).The expression of AKr7a3 gene,ugt2b37 gene,GSTA5 gene,Fmo1 gene,Aldh1a3 gene and Cyp2f4 gene in each group decreased with the increase of DEHP concentration,and the difference was statistically significant(P<0.05).6 The results of Western Blot showed that there was no difference between the AKr7a3 groups,and the difference was not statistically significant.The results are not consistent with the results of quantitative PCR analysis,and the toxic relationship between AKr7a3 gene and DEHP induced embryonic development remains to be further studied.The concentration and expression level of DEHP and Sult2a1 gene,the Maob gene in a dose-dependent manner,the drug group(high dose group)the expression level of Sult2a1 gene and Maob gene were higher than the normal control group,the difference was statistically significant(P<0.05).As the concentration of DEHP increased,the expression level of Sult2a1 gene and Maob gene increased correspondingly in each group.The results were in agreement with the results of quantitative PCR analysis.Conclusions DEHP exposure can cause growth retardation and deformity in embryonic rats,,congestion,edema,and cell apoptosis in placenta.Analysis of gene expression profile showed that DEHP exposure could lead to changes of gene expression in rat placental tissues.The m RNA and protein levels of AKr7a3,ugt2b37,GSTA5,Sult2a1,Fmo1,Maob,Aldh1a3,and Cyp2f4 decreased with the increase of DEHP concentration,while the expression levels of Sult2a1 and Maob increased with the increase of DEHP concentration.