Evaluation of the oxidative stress using biomarker malondialdehyde in atretic folicules of Brazilian snakes from Bothrops genus.

Atresia is a process of cell death or apoptosis of the majority of ovarian follicles, known by a variety of morphological or pathological conditions. A large number of atretic follicles was released by many Bothrops spp. captive females snakes, in the Laboratory of Ecologia and Biological Museum, from Instituto Butantan. Lipoperoxidation induces tissue damage, related to the pathogenesis of distinct diseases, which can lead to cell death. We investigated which biochesmistry mechanisms might be responsible for the release of those atretic follicles doing analises of lipidomic and lipoperoxidation. About 150 mg of fatty tissues from atretic follicles were macerated and washed in PBS+ buffer. Extraction and purification of the lipids were done by the method adapted from Bligh & Dyer. The reaction method of the malondialdehyde biomarker (MDA), a secondary product of lipid peroxidation complexed with thiobarbituric acid, was used. The reading was done at 532 nm. Analyzing the results, a great variation in MDA concentration (from 2.5 to 145.5 nmols/g of lipid) was observed in different atretic follicles of the same pit viper specimen, of the golden lancehead Bothrops insularis. As well as B. jararaca (2.3 to 100.6 nmols/g) and B. leucurus (11.8 to 182 nmols/g). However, in a pairwise comparisons, using a Tukey's test, B. leucurus presented high oxidative stress when compared to the other species (> 50 nmols / g). Lipoperoxidation showed a high content of MDA and of oxidative stress in snakes that alter the mitochondrial membrane and consequently leading to oxidative damage that could induces apoptosis (pro-apoptotic) or control cell proliferation and may induce the repair of this damage. The lipidomic analysis (in progress) it will be possible to verify the lipid composition of the follicles (content and type). Cell cycle analysis will also be performed to check cell death or abortion of these follicles.