Bcl-2 expression in a diabetic embryopathy model in presence of polyamines
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The frequency of congenital malformations is 3–5 times higher in mothers with pregestational diabetes mellitus than in general population. Apparently, this problem is due to change in the expression of apoptotic and antiapoptotic genes induced by the oxidative stress derived from the diabetes/hyperglycemia. One of these genes is Bcl-2, which is associated with the control and inhibition of apoptosis. The purpose of the present work was to study the effect of polyamine addition over expression of Bcl-2 gene in a model of diabetic embryopathy. For this, gestational day 10.5 (GD10.5) rat embryos were incubated at 37°C for 24 h in control medium, medium with high glucose, or medium with high glucose and supplemented with spermidine or spermine. Post-cultured embryos were harvested and observed to obtain morphological scores; some of them were subjected to molecular biology studies: DNA isolation plus conventional PCR or RNA isolation plus RT-PCR; other embryos were fixed with paraformaldehyde and used for immunohistochemical detection of Bcl-2 protein. Although Bcl-2 mRNA was similarly expressed in all rat embryo treatments, Bcl-2 protein was found only in control-incubated embryos. In conclusion, it seems that the inhibition of Bcl-2 gene expression induced by glucose was not reversed by polyamines.
KeywordsPolyamines Cultured rat embryos Bcl-2 expression
M. Sc. Fernando Barrón Moreno (Bioterio, FESI) helps to authors in the care and maintenance of experimental subjects. M. Sc. Alejandro C. Monsalvo (Sequentiation Unit, UBIPRO) made the sequentiation of the PCR products. Authors should acknowledge to P. de B. Liliana Berenice Ramírez Domínguez for grammar revision, and DG Raquel Flores Olmedo for image edition.
This work was partially supported by PAPIIT (IN215414) from DGAPA, UNAM to MPM; and PAPCA 2016 (Project FESI-DIP-PAPCA-2016-17), from the División de Investigación y Posgrado, FES Iztacala, UNAM, to MPM and RMZ.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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