Study Of The Reproductive And Developmental Toxicity Of F1Rats Treated By DBP In Utero And Human Exposures To PAEs

[Objective]①To observe the effects of intrauterine exposure to DBP on the developmental condition of Fl generation rats and reproductive system of both mother rats and mature F1rats;②Metabolomics method was applied to distinguish different DBP doses by analyzing the metabolites of rats;③To estimate the possible human exposure to phthalates in cosmetics;④To predict the possible human exposure to phthalates in plastic bottled beverages and water.[Method]①During the critical period of sexual differentiation in rats (from GD14to GD19), the pregnant rats were administered DBP in various doses(20mg/kg,100mg/kg and500mg/kg)by gavage, assessed body weight of pregnant rats, litter size and sex ratio of Fl rats, etc.; on postnatal days (PND)7and26, two male and female F1rats of each were put to death and the wet weights of the ovaries, livers and kidneys were recorded and organ coefficients were calculated respectively; on PND22, blood samples were collected from the apical part of heart in mother rats;②Enzyme linked immunosorbent assay(ELISA)were applied to detect the hormones level in serum, including estrogen two alcohol (E2), luteinizing hormone (LH), follicle stimulating hormone (FSH), prolactin (P)and testosterone (T);③Observed the histopathological changes of the ovaries of F1female rats and mother rats, the testicular histopathological changes of F1male rats and anogenital distance;④All of the metabolites were detected by UPLC-MS/MS and PCA was used to obtain the differences in serum metabolic profiles of the different DBP exposure doses;⑤Design a questionnaire to investigate the frequency and volume of costimetic use from242women lived in Shanghai and Xian;⑥The phalates levels in cosmetics, beverages and water were determined by Gas chromatography (GC)so that we can estimate the possible human exposure to phalates through cosmetics use and drinks.[Results]①The results show that the experimental doses of DBP exerted no significant effects on various organs of mother rats, but significant effect on serum hormone levels; compared with the control group, serum T levels in the groups treated with100mg/kg and500mg/kg were significantly decreased (P=0.040, P=0.002), while FSH level increased in500mg/kg dose group (P=0.009); with the increase of DBP exposure doses, the weight gain of mother rats during gestational period decreased significantly (F=3.07, P=0.043);②For the F1female rats, on PND7, the serum LH and T levels decreased with DBP exposure doses increasing (F=3.728, P=0.045and F=4.413, P=0.029); on PND26, the E2levels of the female pups decreased significantly (F=6.773, P=0.002), the FSH levels of rats in dosed group(100and500mg/kg)were significantly lower than the control group (P=0.021and P=0.001), and T level in500mg/kg DBP dosed group increased (P=0.049);③For the Fl male rats, on PND7, LH levels increased with the dose of DBP (F=12.764, P<0.001)while T levels decreased sharply (P=0.043), and E2levels were significantly increased (P=0.031)in group treated with DBP dose of500mg/kg; on PND26, compared with the control group, E2levels decreased in all DBP-treated groups (F=5.716, P=0.006), FSH levels decreased (P=0.044)in group treated with500mg/kg DBP and LH levels increased (P=0.005)in100mg/kg group; the indexes in testes of PND7rats significantly decreased in all DBP-treated groups (F=4.709, P=0.006)while grown to PND26, the differences are not apparent; The anogenital distances of PND7rats were shorter after treated with500mg/kg DBP (P=0.025)and this differences still exsited on PND26male rats (P<0.001);④It can be seen from the metabolic profiles of PND7rats, metabolomics technology can be a good method to distinguish the dosed groups from control group, even the low dosed group, there are some differences in the metabolic profiles compared with the control group;⑤According to the result of242-women survey, there were more than70percent of people use the cleanser and lotion about one time per day;75.2%women would use hair products every2days; about40%women use perfume with frequency of2times per week; only10.7percent of women use Deodorant approximately once a week;32.6%women consume nail polish with low frequency of one times two months; DEP、DBP and DEHP in cosmetics were determined, it showed that40%cosmetics were found to contain the DEP with the average concentration of3.775μg/mL, and the detection rate of DBP and DEHP were83.6%and87.8%with concentration of0.043μg/mL and0.059μg/mL respectively; Daily human exposure levels to phthalates via cosmetics were estimated by combining the data of cosmetic use and phathalates concentration in cosmetics, median exposure levels to phthalates in cosmetics by dermal absorption were estimated to be1.119μg/d for DEP,1.112μg/d for DBP and0.538μg/d for DEHP;⑥We have determined3phathalates in51different brands of beverage including functional drinks, coffee drinks and milk drinks; DEP wasn’t found while the detection rates of DBP and DEHP in samples were100%and98% respectively with the average concentrations of0.038mg/L and0.022mg/L respectively; and there were19samples of plastic bottled water, detection rates of DBP and DEHP were42.1%and100%with an average concentration of0.015mg/L and0.019mg/L respectively; Multiple linear regression analysis indicated that compared with drinking water, the reciprocal values of DBP concentration were59.598units lower (p<0.001)in functional drinks,67.492units lower in milk drinks (p<0.001)and83.413units lower in coffee beverages (p<0.001); the reciprocal values of DEHP in coffee beverages were18.980units lowere than in drinking water (P=0.002).[Conclusion]①Intrauterine exposure to DBP exert significant effect on the reproductive system of F1rats even the low dose of DBP can lead to changes in the serum hormone levels, which can be illustrated by the various metabolic profiles;②Most of women consume cosmetics in Shanghai and Xian while it didn’t pose a great threat to the amount of human exposure to phthalates because of the low detected level of phthalates and small volume of cosmetic consumed;③It can be concluded that the detection rates of DBP and DEHP in plastic bottled drinks and drinking waters were fairly high and their levels varied from the different kinds of drink; Due to the low volume of drinks consumed and the detected concentration within the limits of state regulations, plastic bottled beverages are not the the main way of human exposure to phthalates; Since the daily consumption of drinking water is much higher than the drink, the pollution of DBP in drinking water need our more attention.