Astrocytes play a pivotal role in CNS detoxification pathways, where glutathione (GSH) is involved in the elimination of oxygen and nitrogen reactive species such as nitric oxide. We have previously demonstrated that the specific activity of ␥-glutamyl transpeptidase (␥-GT), an enzyme of central significance in GSH metabolism, is regulated in vivo in astrocytes by 1,25-dihydroxyvitamin D 3 (1,25-D 3 ). The aim of the present work was to investigate, in primary cultures of newborn rat astrocytes, the effects of this hormone on ␥-GT synthesis and on GSH and nitrite levels after lipopolysaccharide (LPS) treatment. This study demonstrates that both ␥-GT gene expression and specific activity, induced by LPS, are potentiated by 1,25-D 3 . In contrast, 1,25-D 3 does not regulate the expression of other enzymes involved in astrocyte detoxification processes, such as superoxide dismutase or GSH peroxidase. In parallel, 1,25-D 3 enhanced intracellular GSH pools and significantly reduced nitrite production induced by LPS. Taken together, these results suggest that ␥-GT, GSH, and 1,25-D 3 play a fundamental role in astrocyte detoxification pathways.

The transport and uptake of the most common Se compounds, selenate (SeO42-), selenite (SeO3(2-)), selenomethionine, and selenocystine, were investigated using confluent monolayers of Caco-2 cells, a human carcinoma cell line. Comparative measurements were performed in the absorptive (apical to basolateral side) and exsorptive (basolateral to apical side) directions. Apparent permeability coefficients (Papp), calculated from transport experiments in the absorptive direction, showed increasing values in the following rank order: about 1 x 10(6) cm/s < mannitol < SeO3(2-) < or = selenocystine < selenomethionine < SeO4(2-) < or = about 16 x 10(4) cm/s. The ratios of the Papp measured in the absorptive versus exsorptive directions indicated that only the organic forms presented a net polarized transport (Papp ratio >> 1), suggesting the presence of a transcellular pathway. No significant excretion was observed. The transport of selenomethionine was inhibited by its sulfur analog, methionine, suggesting a common transport mechanism. In contrast, an inhibition of the transport of selenocystine by cysteine was not observed. From the two substrates tested, sulfate and thiosulfate, only thiosulfate inhibited the transport of SeO4(2-) . This effect was also observed for SeO32- (i.e., was unspecific), which questioned the assertion of a common transport for sulfate and SeO4(2-) and may confirm the paracellular pathway of SeO42- suggested by the Papp ratio of about 1. The addition of glutathione (GSH) in large excess had no consequence on the passage of SeO3(2-) but strongly increased the uptake (about fourfold). The liquid chromatography - mass spectrometry (LC-MS) data showed that, in the ionic condition of incubation medium, GSH promptly reduced SeO3(2-) (< or = 2 min) in its elemental form Se0, which cannot ascribe to selenodiglutathione a direct role in the effect of GSH.

The uptake and transport kinetics of manganese (Mn) were investigated in the human intestinal Caco-2 cell line both from the absorption side (apical to basolateral) and from the exsorption side (basolateral to apical). With regard to the former, transport versus time revealed (as uptake) a biphasic pattern with an initial transient phase followed by steady-state conditions. Uptake versus Mn concentrations showed saturation-type kinetics with a 100% increase of Mn binding capacity when measurements were made from 0.5 to 2 h of incubation. The transport characteristics in steady-state conditions exhibited two components, saturable (Vmax = 3.70+/-0.07 nmol/cm2/h, K(m) = 32.2+/-3.4 microM) and nonsaturable (slope = [1.4+/-0.2] x 10(-6)cm(-2)/h) usually presumed to reflect transcellular (carrier mediated) and paracellular (diffusional) pathways, respectively. Mn fluxes were decreased by calcium and calcium antagonists, almost 100% inhibited at 4 degrees C, and affected by quinacrine and ouabain. The inhibition of ATP synthesis was apparently ineffective. From the exsorption side, the Mn fluxes, without a transient period, had an approx 20-fold smaller rate than in the absorptive direction and showed mainly a nonsaturable route (slope = [0.6+/-0.1] x 10(-6) cm(-2)/h). The mechanisms participating in the Mn movements through the monolayer are discussed and proposed to be in common, at least partly, with other divalent cations such as calcium, zinc, or iron.

No abstract

The effects of thyroparathyroidectomy (TPTY) and of replacement therapy using thyroxin (T4) and calcitonin (CT) on the tissue distribution of elements were studied in the rat under semichronic conditions. The elements Na, K, Ca, Mg, Fe, S, P, Rb, Sr, Mn, Cu, and Zn were determined in whole blood, plasma, brain, liver, heart, kidney, skeletal muscle, and bone. TPTY modified concentrations of all elements tested but only small changes were observed for K, Mg, S, and P. The mineral bone composition was slightly modified, 28 d after TPTY, whereas plasma was the most altered. The consequences of TPTY were corrected fairly well by T4 for Na, Cu, Zn, Fe, and S, and by CT for K, P, Rb but with less efficiency for Ca. This study revealed that hormones of the thyroid gland, mainly T4, play an important role in the plasma and tissue balance of elements. It is suggested that T4 participates in tissue fixation of Cu, Zn, and Fe and that CT influences phosphoremia and cellular Ca binding.