Study Of Di (2-Ethylhexyl) Phthalate (DEHP) On Reproductive Toxicity In Mice

Di(2-ethylhexyl)phthalate(DEHP)is a plasticizer.It enters the human body through the food chain and accumulates in the body,which has a serious impact on human reproductive health and has a serious harm to reproduction,development,hormone secretion and gene expression.Previous studies have shown that DEHP can lead to male sterility in offspring,and then cause damage to reproductive ability,and then affect the growth and development of offspring.At the same time,DEHP also has endocrine disrupting effect,affecting the secretion and function of various hormones in the body.Its blood and urine metabolites are closely related to the pathogenesis of diabetes.However,research on reproductive function is still lacking.Although several studies have demonstrated that DEHP is clearly hazardous to development and reproduction and has teratogenic effects,most studies are limited to the animal level and cell level,and lack of population-related epidemiological data.In terms of reproductive toxicity,the toxicity of its female and male reproductive systems was compared,especially the discussion of its toxicity mechanism was not comprehensive enough,and further research was needed.At present,some studies on the mechanism of its toxicity still remain in theory and need more experiments to verify.With the more and more extensive application of DEHP in the population and the more and more exposed people,a systematic study of the environmental risks of DEHP is urgently needed.DEHP is a well-known environmental endocrine substance that can cause reproductive damage.Exposure to environmentally relevant toxins during pregnancy is likely to produce a toxicity quite different from that of the physiological period.In this study,the animal model of the mouse can be used to study the effects of gavage administration of both DEHP and corn oil on reproduction and the growth and development of the offspring.Histopathological analysis,immunohistochemistry,WB,RNA-seq and RT-PCR techniques allow the assessment and determination of the effects of DEHP at different concentrations of exposure on various types of indicators in ICR pregnant mice and their offspring,based on pictures and data.Through the research of this project,it is expected to reveal the effects of DEHP on female mice under specific physiological conditions,and provide reference ideas for its reproductive toxicity research.Compared with the control group,the results showed that:1)Gradient dose exposure to DEHP can relatively significantly reduce the body weight of neonates produced by ICR pregnant rats,their own ovarian,uterine and placental organ factors;2)DEHP exposure significantly improved the efficiency of absorbing fetuses and abnormal fetuses,but there was no significant difference,which increased the morphological changes of uterine tissue;3)The results of HE staining showed that DEHP exposure increased the number of atresia follicles and primordial follicles in ovarian tissue;DEHP causes uterine cavity adhesion or expansion,endometrial congestion and edema,inflammatory cell infiltration and other pathological changes in pregnant mice;DEHP significantly reduced the placental cavernous trophoblast;4)According to IHC and WB,DEHP exposure significantly increased the content of CC-3 and decreased the content of PCNA in the ovary,which was relieved after MLT;DEHP exposure increased the content of CC-3 in uterus,decreased after MLT,increased the content of PCNA,and continued to increase after MLT;DEHP exposure reduced the content of CC-3 and PCNA in placenta,and increased after MLT,without significant effect;5)Through RT-PCR,it can be seen that DEHP exposure has certain inhibition on the expression of transcription factor Nrf2,which indirectly inhibits the expression of reductive enzyme-related genes that should stimulate protection against oxidation,thus producing toxic phenotype.The therapeutic effect of MLT may depend on the metabolism of intermediates through glycolysis.To sum up,the toxicity of DEHP in the ovaries of pregnant mice is mainly caused by oxidative stress,while the therapeutic effect of MLT requires the help of both antioxidant enzyme system and glycolysis.6)According to the WB results,LDHA is the key enzyme in the glycolysis process.LDHA at high levels of expression can contribute to the invasion and migration of malignant cells.The uterus,placenta,trophoblast and tumor cells of mammals with high proliferation and invasion can avoid the host immune response.In the uterine and placental tissues of ICR pregnant rats,LDHA protein expression levels were significantly lower in the DEHP-exposed group than in the control group,but after concomitant treatment with MLT intraperitoneal injection,a significant increase in LDHA protein expression levels was observed,demonstrating that the phenotype could play a therapeutic role.There was no difference in ovary.7)According to the RNA-seq results,the toxic effects produced in the uterus of pregnant mice are mainly related biological processes in the process of steroid synthesis and cholesterol production.These results showed that exposure to DEHP during pregnancy would have a great negative impact on their reproductive ability.The exposure of DEHP and the treatment of MLT are related to the oxidative stress and glycolysis process in pregnant mice.It is speculated that the toxicity of DEHP is related to the oxidative stress and glycolysis energy metabolism pathway.Finally,this study found that melatonin can reverse the female reproductive toxicity induced by DEHP during pregnancy and improve the adverse pregnancy outcome.Melatonin may be a promising drug for alleviating environmental toxicity.