Abstracts / Toxicology Letters 229S (2014) S40–S252 S243

RPTEC/TERT1 cells represent a well differentiated, non-tumorous, proximal tubule-specific phenotype and form transport-ing monolayers with extremely stable gene expression allowingfor long term investigations (weeks to months). Here, we exam-ined the relevant proximal tubule-specific transport characteristicsof RPTEC/TERT1 cells for xenobiotic handling. We demonstratethat these cells express several members of the organic anion andcation transporter family (OATs and OCTs) on a mRNA, protein andfunctional level. The fluorescent OCT substrate, 4-di-1-ASP, waspreferentially taken up at the basolateral side. The nephrotoxincisplatin was transported from the basolateral to the apical sideand basolateral application demonstrated an enhanced toxicitycompared to apical exposure. Furthermore, the OAT substrates phe-nol red and p-aminohippurate were also basolaterally to apicallytransported. Functional expression of p-glycoprotein (MDR1) bycyclosporine A (CsA) inhibition of calcein-AM efflux was demon-strated. Repeat dose exposure to CsA over 14 days resulted in anon-linear cellular accumulation. Finally, long-term repeat doseexposure to several other clinical relevant nephrotoxins demon-strates the suitability of these cells for chemical safety assessmentand biomarker development.

http://dx.doi.org/10.1016/j.toxlet.2014.06.809

P-4.122Does developmental exposure to bisphenol Ainduce bone and adipose tissue disturbances?

Margareta Halin Lejonklou 1,∗, Elina Karimullina 2, Sune Larsson 3,Thomas Lind 1, Håkan Melhus 1, Annica Jacobson Rasmusson 1,Monika Rönn 1, Ulf Risérus 4, Bruce Blumberg 2, P. Monica Lind 1

1 Department of Medical Sciences, Uppsala University, Uppsala,Sweden, 2 Department of Developmental and Cell Biology, Universityof California, Irvine, Irvine, California, USA, 3 Department of SurgicalSciences, Uppsala University, Uppsala, Sweden, 4 Department ofPublic Health and Caring Sciences, Uppsala University, Uppsala,Sweden

Background: Bisphenol A (BPA) exposure has been reported tobe associated with obesity, and hormone disruption with alteredbone density and mineralization.

Aims: We evaluated effects of Bisphenol A exposure during ges-tation and lactation, in five-week-old female and male Fischer F344rats, on bone and adipose tissue development in a pilot study todetermine doses in our ongoing main study.

Methods: Pregnant Fischer F344 rats were exposed to BPA fromgestational day 10.5. Adipose tissue depots and liver from offspringwere collected for fatty acid analyses and liver from dams for ultra-structural examination. Bone marrow stem cells were isolated forex vivo cell culture and subsequently an adipogenesis assay. Bonewas collected for bone density measurements, using pQCT-analysis.

Results: Gestational low and medium level exposure (0.5 and5 �g/kg bw/day, respectively) to BPA significantly induced adi-pogenesis in ex vivo culture of F344 bone marrow mesenchymalstem cells. Interestingly, the presence of PPAR activator in theinduction cocktail was essential and significantly increased lipidaccumulation in cells harvested from low- and medium-high doseBPA treated animal groups. The fatty acid composition in liver aswell as adipose tissue in offspring was altered in BPA-exposedanimals. Liver ultrastructural disturbances were observed in low-and high-dose exposed dams compared to control. Furthermore,pQCT results revealed that exposure to BPA increased trabecularbone in females and decreased cortical bone in males. Conclusion:These data support the hypothesis that perinatal low-dose BPA

exposure may predispose to obesity development, and also affectbone formation.

http://dx.doi.org/10.1016/j.toxlet.2014.06.810

P-4.123Bisphenol A increases cortisol production byenhancing phosphorylation of CREB in normalhuman adrenocortical cells

Margareta Halin Lejonklou 1,∗, Per Hellman 2, Johan Botling 3, P.Monica Lind 1, Peyman Björklund 2

1 Department of Medical Sciences, Uppsala University, Uppsala,Sweden, 2 Department of Surgical Sciences, Uppsala University,Uppsala, Sweden, 3 Department of Immunology, Genetics andPathology, Uppsala University, Uppsala, Sweden

Background: Involvement of bisphenol A exposure in obesityhas been demonstrated before. Abnormal cortisol production isinvolved in aberrant fat distribution in humans. CREB phosphory-lation is a major regulatory event in cortisol production. The effectsof bisphenol A on cortisol production in adrenals is not completelyunderstood.

Methods: Primary cell cultures from human adrenal tissue weretreated with 1 �M bisphenol A and secreted and total cortisol weremeasured by using ELISA. Western blotting analysis was performedusing a phospho-specific antibody.

Results: Secreted and total cortisol were increased (2.1× and3.8×; p < 0.05, respectively) after treatment of human primaryadrenocortical cells with bisphenol A in the presence of recombi-nant human ACTH. CREB phosphorylation was significantly higherin cells treated with bisphenol A (6.2×; p < 0.05).

Conclusion: Bisphenol A might contribute to aberrant fat distri-bution by upregulation of cortisol production.

http://dx.doi.org/10.1016/j.toxlet.2014.06.811

P-4.124Effects of ferulic acid on oxidative stressparameters in livers and kidneys of Wistaralbino rats

Merve Bacanli 1, Sevtap Aydin 1, Gökce Taner 2, Hatice GülGöktas 1,3,∗, Tolga Sahin 4, A.Ahmet Basaran 5, Nursen Basaran 1

1 Hacettepe University, Faculty of Pharmacy, Department ofToxicology, Ankara, Turkey, 2 Gazi University, Faculty of Science,Department of Biology, Ankara, Turkey, 3 Cukurova University,Faculty of Pharmacy, Department of Toxicology, Adana, Turkey,4 Kastamonu University, Faculty of Medicine, Department of Surgery,Ankara, Turkey, 5 Hacettepe University, Faculty of Pharmacy,Department of Pharmacognosy, Ankara, Turkey

Oxidative stress has an important role in the developmentof sepsis-induced multiorgan failure. Ferulic acid (FA), a well-established natural antioxidant, has several pharmacologicalactivities including anti-inflammatory, anticancer and hepatopro-tective. The aim of this study is to investigate the protective effectsof FA against sepsis-induced oxidative stress. Rats were divided intofour groups. Two groups served as sham and sepsis control. Onegroup was treated with FA (100 mg/kg) and the other group wassubjected to sepsis and treated with FA (100 mg/kg). The oxidativestress parameters such as superoxide dismutase (SOD), glutathioneperoxidase (GSH-Px) activities and malondialdehyde (MDA), totalglutathione (GSH) and tumor necrosis factor-� (TNF-�) levels were