Ethylparaben Effects On Drosophila Melanogaster Estrogen-related Gene Expression

P-hydroxybenzoic acid esters, known as enhanceparaben, are used in food, cosmetics, chemical, medicine and various industrial corrosion fields, because of its broad antimicrobial spectrum, low cost, high security, stability to acid-base, etc. All this time, parabens have been supposed to be a kind of safe and effective preservative. However, preservatives can enter the body through a variety of ways, and can accumulate in the body. A lot of studies proved that it can affect the animal’s reproductive organs, and has adverse effects on human breast cancer cells.Drosophila melanogaster is a ideal model organism in Genetics and other research fields, because it is characterized by small size, rapid growth, shorter life cycle (about two weeks), strong fecundity, convenient feeding, low cost, less chromosome number, more mutant charater, etc. Futhermore, fruit fly genes have sequence homology with human genes,so it has many advantages than other creatures in reproduction and aging aspect study.In this experiment, the Drosophila melanogaster was used experimental meterals to study the impact of the EP to its physiological and molecular level. We collected the male and female virgin that placed in different concentrations of EP (0.00%,0.02%,0.10%and0.20%) to study the lifespan and its fecundity. Furthermore, we discussed the estrogen influence of EP on the expression of ERR, EcR and YPR after feeding ten days, twenty days, thirty days, forty days,fifty days and sixty days at the molecular level by Quantitative real-time PCR.The results show:1. Culture medium adding0.00%,0.02%,0.10%and0.20%respectively in the concentration of EP, certain changes has taken place in life indicators of fruit flies. Results show that the average life span for the male and female fruit flies, half the time of death, the highest life were lower than control group (p<0.05), and showed a certain dose-response effect. The average life span of the0.20%group was46.18±1.53day and42.39±1.66day. The results showed that EP influenced the lifespan of flies.2. Compared with the control group, the minimum concentrations (0.02%) of female flies produce increased significantly the number of Drosophila produced offspring (p<0.05), and the concentrations of EP (0.10%,0.20%) reduced the number of hatching in flies. It indicated that a certain concentration of EP can lead to the weakening of Drosophila fertility.3. Compared with the control group, the minimum concentrations (0.02%) of flies were shortened the emergence period of its offspring significantly, and the0.10%,0.20%of flies were prolonged eclosion time of its offspring. It showed that EP impacted the growth cycle of Drosophila.4. Influence of EP on the expression of ERR, EcR and YPR after feeding ten days, twenty days, thirty days, forty days,fifty days and sixty days at the molecular level by quantitative real-time PCR. The results showed that ERR in all the different concentration of EP were induced observably at thirty days and forty daysfor females. EP treatment (0.10%) tended to increase the expression ERR high significantly (p<0.01) in the short time process,(twenty days). In addition, high concentration EP treatment (0.20%) tended to increase the expression ERR significantly in the short time treatment (twenty days) for males, and0.02%of EP tended to increase the ERR significantly (p<0.05) in the long time treatment (fifty days and sixty days).5. The0.20%EP upregulated the EcR expression of Drosophila significantly compared with the control after exposure ten days, twenty days and thirty days. The EcR in0.02%of EP was induced observably at forty days, fifty days and sixty days, and the EcR in0.10%of EP tended to increase the expression high significantly (p<0.01) at fifty days.. In male, the0.10%and0.20%of EP were induced the expression EcR after ten days, twenty days and thirty days. The increase magnitude of female flies is higher than the male flies.6. Compared with the control, in female, high concentration EP treatment (0.20%) tended to increase the expression YPR significantly (twenty days, thirty days and forty days), and the0.10%of EP tended to increase the expression YPR significantly (fifty days and sixty days). The YPR in0.02%of EP were induced observably at sixty days (p<0.01). In male, the0.10%and0.20%of EP were induced the expression YPR (thirty days, forty days and fifty days). The increase magnitude of female flies is higher than the male flies.Considering various research results, the four concentrations of EP on Drosophila melanogaster lifespan, fecundity and the effect of ERR, EcR and YPR. The above data clearly confirm the toxicity and potential estrogenic effects of EP.