Efeito da adição de Glutationa na função e estresse oxidativo em sêmen ovino criopreservado / Effect of glutathione on ovine cryopreserved sperm function and oxidative status

AUTOR(ES)
DATA DE PUBLICAÇÃO

2009

RESUMO

It is well known that sperm is extremely susceptible to the oxidative stress. This occurs especially due to the high content of poly-unsaturated fatty acids (PUF As) in its plasma membrane. The PUF As provide the necessary fluidity to the plasma membrane, which allows the sperm to move. However the double bonds present in those fatty acids are more susceptible to the oxidative stress. Therefore, sperm is highly dependent of the extra-celluIar environment antioxidant protection. Studies in human indicate that cryopreservation may lead to damages to the sperm due to the oxidative stress. The objective of this experiment was to study if the antioxidant glutathione (GSH) may protect ovine cryopreserved sperm against the damages caused by the oxidative stress. Semen samples of four rams were collected using an artificial vagina. Samples were then cryopreserved using Tris-egg yolk extender supplemented with different concentrations of reduced glutathione (control, 1, 5 and 10 mM). After thawing, samples were evaluated using conventional (motility and vigor) and functional tests (membrane integrity and mitochondrial activity). An aIiquot of each thawed sample was submitted to the protocol of induced lipid peroxidation using ascorbate (20 mM) and ferrous sulphate (4 mM), with the further measurement of the tiobarbituric acid reactive substances (TBARS), as an index of oxidative stress. This technique aims to evaIuate sperm oxidative status by its susceptibility to the lipid peroxidation. Statistical analysis was performed using SAS system for Windows. The results indicate that there was no effect of GSH on the variables assessed by conventional tests. GSH decreased the proportion of intact acrosomes. Samples treated with 5 mM GSH showed a lower percentage of cells with intact membranes when compared with control sampIes and those treated with 10 mM, the percentage of cells with mitochondriaI activity was affected by GSH and there was also no effect on TBARS. Samples from the control group were more susceptible to denaturation of chromatin. In conclusion, the addition of the antioxidant Glutathione (GSH) offers protection to DNA and mitochondrial activity of ovine sperm.

ASSUNTO(S)

criopreservação cryopreservation free radicais giutathione glutationa ovine ovino radicais livres semen sêmen

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