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Favaro RR. Establishment of a mouse model of pregnancy complicated by type 1 diabetes: Evaluation of its impact on the uterine environment at early pregnancy. [Ph. D. thesis (Tecidual and Cellular Biology)]. São Paulo: Instituto de Ciências Biomédicas da Universidade de São Paulo; 2011. By means of this study our laboratory has developed a new mouse model of pregnancy complicated by long-term type 1 diabetes and evaluated the impact of this disease on the uterine environment at early pregnancy. The age of pregnancy studied (168 hours) comprises the stage in which antimesometrial decidua is fully developed and organogenesis is in progress, of particular interest due to higher rates of malformations in diabetic pregnancies. In addition, the myometrium is at the first adaptative phase of pregnancy (proliferative phase) and diabetic pregnancies are associated with high incidence of preterm labors. Diabetes was induced in female Swiss mice by i.v. alloxan administration (40 mg/Kg) and the animals were mated after different periods of time. Diabetic females showed pathophysiological parameters of type 1 diabetes, characterized by hyperglycemia, hypoinsulinemia, polyuria, glycosuria, polyphagy, polydipsia and decreased body weight. Comparison of the number of implantation sites and decidual dimensions between groups mated 50-70 days after diabetes induction (D) or 90-110D showed that both parameters were impaired only in the 90-110D group, which was elected for subsequent studies. Picrosirius staining revealed an augment in fibrillar collagen deposition in the diabetic group, when compared to control group. Immunohistochemical analysis demonstrated increased deposition of collagen types I and V, and decreased deposition of collagen type III, and associated proteoglycans biglycan and lumican. These alterations reflected on collagen fibrillogenesis, as shown by transmission electron microscopy (TEM) analysis. Quantitative real time PCR evaluation of collagens and proteoglycans mRNA expression, however, demonstrated the increase of collagen type I expression only. Taking into account reports on the role of IL-11 on decidual extracellular matrix (ECM) remodeling, its levels were measured by ELISA. However, no differences were observed between groups. Effects of diabetes on the myometrium were investigated in two subgroups: D1 (90-110D) and D2 (100-110D). Histomorphometrical analysis revealed an increase in the distance between muscle layers in D1, indicating edema, and diminution of the distance in D2, associated with atrophy and fibrosis of the myometrium. In both subgroups narrowing of muscle layers was detected and correlated with decreased cell proliferation of smooth muscle cells, evaluated by immunodetection of PCNA. TEM analysis showed impairment of cytoarchitecture, ECM and contractile apparatus of the smooth muscle cells, confirmed by immunolocalization of α-smooth muscle actin. Together, our results support the concept that development of gestational complications is directly associated with the progression of diabetes in the mother, in consonance with clinical results, pointing to the importance of experimental models that include the temporal factor to investigate the effects of diabetes on pregnancy. Additionally, important alterations have been described in this model, which contribute to explain the alterations on embryo development and the high incidence of preterm labor in diabetic pregnancies.

Estabelecimento de um modelo de gestação complicada por diabetes tipo 1 em camundongos: avaliação do seu impacto sobre o ambiente uterino no início da gestação.