University of Colorado, Denver, CO, USA. Gestational Diabetes Mellitus (GDM) is a prevalent disease that negatively impacts fetal and placental development.
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Abstracts / Placenta 36 (2015) 469e521
up with non-pregnant or/and non-infected mice. The animals were evaluated for mortality and body weight loss. Sera were collected and analyzed for determination of cytokines profile by CBA assay. Also, the uteri were evaluated by western blotting for quantification of MIF receptor (CD74) and Indoleamine 2,3 Dioxygenase (IDO). Results: Our results demonstrated that C57BL/6 MIF-/- succumbed to infection before WT. Also, non-pregnant C57BL/6 MIF-/- presented more pronounced body weight loss than WT. Regarding CD74 expression in the uterus, we observed higher expression of the receptor in C57BL/6 MIF-/- than in WT. The analysis of IDO demonstrated that it was expressed in higher amounts in the uteri of C57BL/6 MIF-/- than in WT. Related to the cytokines profile, we observed higher levels of pro-inflammatory cytokines in C57BL/6 WT. On the other hand, when the anti-inflammatory cytokines were analyzed, we observed higher levels of IL4 and IL10 in the serum of C57BL/6 MIF-/-. Conclusions: In conclusion, MIF demonstrated to be important for control of T. gondii infection in pregnant and non-pregnant mice. The absence of MIF in C57BL/6 MIF-/- leads to upregulation of MIF receptor, demonstrating the importance of MIF during pregnancy. The mechanism of MIF action in the maternal-fetal interface may be related to IDO suppression and induction of a pro-inflammatory immune response. Financial support: CAPES, FAPEMIG, CNPq
PB.33. CHLORPYRIFOS INDUCES RETICULUM ENDOPLASMIC STRESS IN JEG-3 CELLS L. Reyna, M.E. Ridano, J.B. Flores-Martín, G. Panzetta-Dutari, S. GentiRaimondi. CIBICI-CONICET, Dpto. Bioquímica Clínica, Facultad de Ciencias rdoba, Argentina Químicas, UNC, X5000HUA, Co Chlorpyrifos (CPF) is an organophosphorous pesticide widely used in agricultural, industrial, and household applications. We have previously shown that JEG-3 cells are able to attenuate the oxidative stress induced by CPF through the adaptive activation of the Nrf2/ARE pathway. Particularly, we found that cells treated with 50 ìM or 100 ìM CPF lead to an early Nrf2 increase at both mRNA and protein levels, triggering the antioxidant status and nuclear Nrf2 translocation. Objectives: This work was performed to evaluate whether CPF induces reticulum endoplasmic (ER) stress in JEG-3 cells. Methods: Cells were exposed to 50 ìM or 100 ìM CPF for 3 or 24 h in conditions where cell viability was not altered. Western blot assays were used to explore the protein levels of ER stress biomarkers. Results: In response to CPF the ER stress biomarker proteins glucoseregulated protein 78 and calnexin were significantly augmented at 24 h of treatment, whereas protein disulfide isomerase showed an increasing trend although statistically non-significant. Furthermore, CPF also activated inositol-requiring enzyme 1, a sensor for the unfolded protein response. Since reported studies indicate that p53 plays a critical role in the cellular response to ER stress, the effect of CPF was investigated by measuring p53 protein expression. The data indicate that CPF induces an early p53 expression at 3 h of CPF cell exposure which was recovered to the basal value at 24 h of CPF treatment. Conclusions: Altogether, these findings indicate that CPF induces ER stress in JEG-3 cells; however they are able to trigger an adaptive protection response against the xenobiotic injury. Supported by CONICET, FONCyT and SECyT-UNC
PB.34. ACTIVATION OF MTOR SIGNALING AND INCREASED NITRIC OXIDE METABOLISM IN THE PLACENTA OF RATS WITH GESTATIONAL DIABETES E. Capobianco 1, V.I. Ramirez 2, D. Fornes 1, T.L. Powell 3, T. Jansson 4, A. Jawerbaum 1. 1 Laboratory of Reproduction and Metabolism, School of Medicine, CONICET-CEFYBO-UBA, Buenos Aires, Argentina; 2 Center for Pregnancy and Newborn Research, University of Texas Health Science Center, San Antonio, TX, USA; 3 Department of Pediatrics, University of Colorado, Denver, CO, USA; 4 Department of Obstetrics & Gynecology, University of Colorado, Denver, CO, USA
Gestational Diabetes Mellitus (GDM) is a prevalent disease that negatively impacts fetal and placental development. Mechanistic target of Rapamycin (mTOR) regulates important metabolic pathways related the placental transfer of nutrients and fetal growth. Nitric Oxide (NO) is a vasodilatory agent, and a substrate in the formation of peroxynitrites. Objective: Using a novel model of GDM in the rat, involving female offspring of pre-gestational diabetes or control mothers, we tested the hypothesis that mTOR is activated and NO overproduced in their placentas. Methods: Pregestational diabetes was generated by neonatal streptozotocin administration (F0). Female diabetic and control rats were mated with control males. Their female offspring (F1) were mated with control males. F1 females born to mothers with pre-gestational diabetes developed GDM. At day 21 of pregnancy, the placentas and fetuses from control and GDM rats were collected and weighed. The expression of proteins involved in mTORC1 (S6K1, RP-S6K1 and 4EBP1) and mTORC2 (SGK and PKCa) signaling pathways, were determined by Western blot in the placenta (total and phosphorylated proteins). Placental nitrates and nitrites (indicating NO production, by Griess reaction) and protein nitrotyrosylation (indicating peroxynitrite-induced damage, by immunohistochemistry) were determined. Results: Fetal weight was higher in the GDM group as compared to control (p
