Mg. Muñoz-Roa, Mirna

The mammary gland increases energy requirements during pregnancy and lactation to support epithelial proliferation and milk nutrients synthesis. Lactose, the principal carbohydrate of the milk, is synthetized in the Golgi of mammary epithelial cells by lactose synthase from glucose and UPD galactose. We studied the temporal changes in the expression of GLUT1 and GLUT8 in mammary gland and their association with lactose synthesis and proliferation in BALB/c mice. Six groups were used: virgin, pregnant at 2 and 17 days, lactating at 2 and 10 days, and weaning at 2 days. Temporal expression of GLUT1 and GLUT8 transporters by qPCR, western blot and immunohistochemistry, and its association with lactalbumin, Ki67, and cytokeratin 18 within mammary tissue was studied, along with subcellular localization. GLUT1 and GLUT8 transporters increased their expression during mammary gland progression, reaching 20-fold increasing in GLUT1 mRNA at lactation (p < 0.05) and 2-fold at protein level for GLUT1 and GLUT8 (p < 0.05 and 0.01, respectively). The temporal expression pattern was shared with cytokeratin 18 and Ki67 (p < 0.01). Endogenous GLUT8 partially co-localized with 58 K protein and α-lactalbumin in mammary tissue and with Golgi membrane–associated protein 130 in isolated epithelial cells. The spatial-temporal synchrony between expression of GLUT8/GLUT1 and alveolar cell proliferation, and its localization in cis-Golgi associated to lactose synthase complex, suggest that both transporters are involved in glucose uptake into this organelle, supporting lactose synthesis.

Tobacco smoke causes oxidative damage directly by the effect of their oxidants or indirectly through the induction of endogenously produced oxidants and/or inactivation of antioxidants. The oxidative effect of tobacco smoke depends on many variables: dose, time of exposure, tissue or cell type and endogenous antioxidant status. In an attempt to simplify this complex scenario, we examined the effect of a soluble extract of tobacco smoke on the activity of purified antioxidant enzymes (catalase, glutathione peroxidase and superoxide dismutase) and in human plasma. Our results revealed that catalase and glutathione peroxidase were inhibited with an IC50 of 18 and 80 smoker equivalents (arbitrary units), respectively; meanwhile superoxide dismutase was not affected. A similar effect of soluble extract of tobacco smoke was obtained for antioxidant enzymes in human plasma, where catalase was inhibited, while superoxide dismutase was little affected, and glutathione peroxidase increased 20% its activity. Benzo[a]pyrene, a well-known component of tobacco smoke, was partly responsible for catalase inactivation. Although soluble extract of tobacco smoke and benzo[a]pyrene both induced carbonylation of plasma proteins, we ruled out that catalase inhibition would be caused by carbonylation, since the inhibition was reversed by dialysis. Considering the higher sensitivity of catalase to inhibition induced by soluble extract of tobacco smoke and its important role in peroxide elimination, we conceived that benzo[a]pyrene and other compounds of tobacco smoke extract promote a transient peroxide accumulation which could be one of the factors responsible for the oxidative damage in respiratory tract and other tissues in smokers.