title: Municipal wastewater affects adipose deposition in male mice and increases 3T3-L1 cell differentiation.
abstract: Trace concentration of EDs (endocrine disrupting compounds) in water bodies caused by wastewater treatment plant effluents is a recognized problem for the health of aquatic organisms and their potential to affect human health. In this paper we show that continuous exposure of male mice from early development to the adult life (140 days) to unrestricted drinking of wastewater collected from a municipal sewage treatment plant, is associated with an increased adipose deposition and weight gain during adulthood because of altered body homeostasis. In parallel, bisphenol A (BPA) at the administration dose of 5 μg/kg/body weight, shows an increasing effect on total body weight and fat mass. In vitro, a solid phase extract (SPE) of the wastewater (eTW), caused stimulation of 3T3-L1 adipocyte differentiation at dilutions of 0.4 and 1 % in the final culture medium which contained a concentration of BPA of 40 nM and 90 nM respectively. Pure BPA also promoted adipocytes differentiation at the concentration of 50 and 80 μM. BPA effect in 3T3-L1 cells was associated to the specific activation of the estrogen receptor alpha (ERα) in undifferentiated cells and the estrogen receptor beta (ERβ) in differentiated cells. BPA also activated the Peroxisome Proliferator Activated Receptor gamma (PPARγ) upregulating a minimal 3XPPARE luciferase reporter and the PPARγ-target promoter of the aP2 gene in adipose cells, while it was not effective in preadipocytes. The pure estrogen receptor agonist diethylstilbestrol (DES) played an opposite action to that of BPA inhibiting PPARγ activity in adipocytes, preventing cell differentiation, activating ERα in preadipocytes and inhibiting ERα and ERβ regulation in adipocytes. The results of this work show that the drinking of chemically-contaminated wastewater promotes fat deposition in male mice and that EDs present in sewage are likely responsible for this effect through a nuclear receptor-mediated mechanism.
authors.affiliation: Laboratory of Biotechnology, Civic Hospital of Brescia, Brescia, Italy; Department of Molecular and Translational Medicine, University of Brescia, Brescia, Italy.
authors.forename: Giorgio
authors.initials: G
authors.lastname: Biasiotto
authors.affiliation: Laboratory of Biotechnology, Civic Hospital of Brescia, Brescia, Italy; Department of Molecular and Translational Medicine, University of Brescia, Brescia, Italy.
authors.forename: Isabella
authors.initials: I
authors.lastname: Zanella
authors.affiliation: Laboratory of Biotechnology, Civic Hospital of Brescia, Brescia, Italy.
authors.forename: Alice
authors.initials: A
authors.lastname: Masserdotti
authors.affiliation: DIMI Department of Mechanical and Industrial Engineering, University of Brescia, via Branze 38, I-25123 Brescia, Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: Pedrazzani
authors.affiliation: DICATAM Department of Civil, Environmental, Architectural Engineering and Mathematics, University of Brescia, via Branze 43, I-25123 Brescia, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Papa
authors.affiliation: Laboratory of Biotechnology, Civic Hospital of Brescia, Brescia, Italy; Department of Molecular and Translational Medicine, University of Brescia, Brescia, Italy.
authors.forename: Luigi
authors.initials: L
authors.lastname: Caimi
authors.affiliation: Laboratory of Biotechnology, Civic Hospital of Brescia, Brescia, Italy. Electronic address: diego.dilorenzo@yahoo.it.
authors.forename: Diego
authors.initials: D
authors.lastname: Di Lorenzo
mesh_terms: D041721:3T3-L1 Cells; D017667:Adipocytes / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D000273:Adipose Tissue / Q000187:drug effects* / Q000378:metabolism; D050154:Adiposity / Q000187:drug effects; D000818:Animals; D001559:Benzhydryl Compounds / Q000633:toxicity; D002454:Cell Differentiation; D052244:Endocrine Disruptors / Q000633:toxicity*; D047628:Estrogen Receptor alpha / Q000378:metabolism; D047629:Estrogen Receptor beta; D005260:Female; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D047495:PPAR gamma / Q000235:genetics; D010636:Phenols / Q000633:toxicity; D011247:Pregnancy; D062065:Wastewater / Q000633:toxicity*; D000098826:Bisphenol A Compounds
chemical_list: D001559:Benzhydryl Compounds; D052244:Endocrine Disruptors; D047628:Estrogen Receptor alpha; D047629:Estrogen Receptor beta; D047495:PPAR gamma; D010636:Phenols; D062065:Wastewater; C025256:nonylphenol; C006780:bisphenol A; D000098826:Bisphenol A Compounds
keywords: Adipocytes differentiation; Adipose deposition; Endocrine disruptors; Wastewater