Morphological analysis of the pancreatic beta cells of diabetic female rats at different ages of life

Abstract

s / Placenta 36 (2015) 469e521 490 pregnancy morbidity plus vascular thrombosis (PM/VT), pregnancy morbidity without aPL (PM/aPL-) or healthy women (NHS). Drugs were added at the same time with IgG at previous established concentrations. After 24 h, the inserts were removed and trophoblast cell migration across the Matrigel coated membrane was determined using a colorimetric assay. The relative cell invasion value was assessed by comparing the optical densities with no treatment control. STAT-3 activation was determined at same conditions of invasion assays using Western blots. Results: IgGs from patients with APS-associated pregnancy morbidity (PM, PM/VT) induce a significantly decreased invasion in comparison with IgGs from the NHS and PM/aPLgroups. Enoxaparin, aspirin and ATL restore the aPL-decreased invasion and, moreover, ATL has a receptormediated effect as demonstrated by application of its antagonist Boc-2, which reverses the ATL-restored invasion. Additionally, phosphorylation of STAT-3 is reduced by IgGs from PM patients. Conclusion: We show here that some of drugs used in APS-related pregnancy morbidity treatment are involved in the modulation of trophoblast function. They possibly restore some processes essential for normal trophoblast invasion. Moreover, ATL effects may explain the benefits of aspirin treatment observed in patients with obstetric APS. Since STAT-3 is an intracellular signalling molecule involved in migration and invasion processes, our results support the view that aPL from patients with pregnancy morbidity alter the proper function of this pathway. Financial support: CODIUniversidad de Antioquia Grant 91515; Estrategia de sostenibilidad Grupo Reproducci on 2014-2015; Programa de Cooperaci on Cientifica Internacional-Proyectos de Intercambio: Colciencias-Colombia Convocatoria 614-2013, y CONICYT-Chile PCCI 130017. AMA is a fellow of Colciencias. PA.7. MORPHOLOGICAL ANALYSIS OF THE PANCREATIC BETA CELLS OF DIABETIC FEMALE RATS AT DIFFERENT AGES OF LIFE F.Q. Gallego , Y.K. Sinzato , B. Dallaqua , I.L. Iessi , R.L. Amorim , C.E. Fonseca , I.M.P. Calderon , M.V.C. Rudge , D.C. Damasceno . 1 Laboratory of Experimental Research on Gynecology and Obstetrics, Graduate Program on Gynecology, Obstetrics and Mastology, Botucatu Medical School, Brazil; Department of Pathology, Botucatu Veterinary and Zootechnology Medical School, Univ. Estadual Paulista_Unesp, Botucatu, S~ ao Paulo, Brazil Background: Recent studies have shown regeneration of pancreatic islets in physiological and pathological circumstances. We hypothesized that diabetes disturbs insulin-producing b-cell regeneration in laboratory animals. Aim: The objective was to understand the proliferation and death mechanism of pancreatic b-cells in non-diabetic and neonatally streptozotocininduced mild diabetic rats at different stages of life and during pregnancy. Methods: The rats were randomly assigned into the following groups: Non-diabetic (Control1⁄4C, glycemia<120 mg/dL) or Mild diabetes (MD) (n1⁄415/group). The mild diabetes group was induced at birth by the b-cell cytotoxic agent e Streptozotocin (STZ 100 mg/kg b.w., sc.). On postnatal days (PND) 5, 15 and 90 and on day 18.5 of pregnancy, the different groups of rats were killed to collect blood samples for insulin and glycated hemoglobin measurements; pancreas samples were processed for histological and immunohistochemical analysis for cell proliferation, apoptosis and insulin. In addition, on day 18.5 of pregnancy, the uterine corns were exposed for evaluation of reproductive outcomes. p<0.05 was considered as the significant statistical limit. Results: On PND5, the diabetic rats showed pancreatic islet morphological alterations as decreased insulin and apoptosis immunostaining, and increased cell proliferation. On PND15, cell proliferationwas lower than on PND5 in diabetic rats. In adulthood (PND90 and pregnancy), diabetic rats presented increased glycated hemoglobin levels and glucose intolerance, and no changes in pancreatic islets morphology were observed in relation to the control group. On day 18.5 of pregnancy, MD dams presented an increased number of embryonic death and rate of preimplantation losses when compared with the control group. Conclusion: Our results showed that after administration of streptozotocin pancreatic b-cells were able to regenerate partially by an increase in proliferation and a decrease in apoptosis. However, this was insufficient to reverse the diabetic status in adulthood leading to impairment of the reproductive function in diabetic mothers. Acknowledgement: FAPESP/Brazil (Process Numbers: 2011/18519-7; 2011/15606-6). PA.8. LEPTIN IS INVOLVED IN HUMAN TROPHOBLAST MIGRATION AND INVASION A. Toro , R. Sampayo , A. P erez-P erez , B. Maskin , V. S anchezMargalet , M. Simian , C. Varone . Dpto. de Quimica Biol ogica, FCENUBA, IQUIBICEN CONICET, Buenos Aires, Argentina; 2 Area de Investigaciones, Instituto de Oncologia “ Angel H. Roffo”, Buenos Aires, Argentina; Dpto. de Bioquimica M edica y Biologia Molecular. Universidad de Sevilla, Sevilla, Spain; Hospital Nacional “A. Posadas”, Buenos Aires,