Maternal exposure to different combined phytalat: transgenerational effects related to the prostatic development

Abstract

Interesting in environmental factors that interfere with intrauterine development and promote morphofunctional, transient, or permanent changes, including those that can be transmitted to the offspring, constitute an area of extreme scientific interest called DOHaD (Developmental Origins of Health and Disease). Phthalates represent a group of substances used in industry and are found in different concentrations in the urine and plasma of humans. Thus, the objective of this study will be to investigate whether gestational and lactational exposure to different concentrations of a mixture of phthalates is capable of altering the prostate structure of F1 and F2 generation rats, as well as of molecular tissue markers capable of demonstrating changes in the development and physiology of the gland. To reach this goal, pregnant rats of the Sprague-Dawley line will be randomly divided into 4 experimental groups: (control (vehicle), 20 ¼g / kg / day, 200 ¼g / kg / day, 200 mg / kg / day) the respective doses of the mixture by gavage in the following proportion: 19% DEHP (Bis (2-ethylhexyl) phthalate), 36% DEP (Diethylphthalate), 15% DBP (Di-n-butylphthalate), 10% DiBP (DiisobutylPhthalate) BBzP (Butylbenzylphthalate), and 10% DiNP (Diisononylphthalate); and will be exposed from gestational day 10 (DG10) to postnatal day 21 (DPN21). The F1 generation males will be euthanized in DPN22 and DPN120. Some F1 males will be mated with non-exposed females and their offspring (F2) will be euthanized in the DPN22. Gestational data, as well as anogenital distance and testicular descent will be evaluated. The ventral prostate of the animals will be weighed, and a hemilobo will be fixed in Methacarn and intended for histological and immunohistochemical analyzes (IHC), while the other hemilobo will be frozen in liquid nitrogen for protein extraction and Western blotting. The following tissue markers for IHC will be evaluated: AR (androgen receptor), Ki67 (cell proliferation); alpha-actin (smooth muscle) and E-cadherin (epithelial junctions); and by WB: SIRT1 (oxidative stress marker) and Prostateine (functional activity marker). This project is part of a larger study with an epigenetic molecular approach that is being developed under the coordination of Prof. Adj. Wellerson Rodrigo Scarano (co-advisor), with international collaborations and FAPESP funding.