Cobalt distribution in keratinocyte cells indicates nuclear and perinuclear accumulation and interaction with magnesium and zinc homeostasis.

Cobalt is known to be toxic at high concentration, to induce contact dermatosis, and occupational radiation skin damage because of its use in nuclear industry. We investigated the intracellular distribution of cobalt in HaCaT human keratinocytes as a model of skin cells, and its interaction with endogenous trace elements. Direct micro-chemical imaging based on ion beam techniques was applied to determine the quantitative distribution of cobalt in HaCaT cells. In addition, synchrotron radiation X-ray fluorescence microanalysis in tomography mode was performed, for the first time on a single cell, to determine the 3D intracellular distribution of cobalt. Results obtained with these micro-chemical techniques were compared to a more classical method based on cellular fractionation followed by inductively coupled plasma atomic emission spectrometry (ICP-AES) measurements. Cobalt was found to accumulate in the cell nucleus and in perinuclear structures indicating the possible direct interaction with genomic DNA, and nuclear proteins. The perinuclear accumulation in the cytosol suggests that cobalt could be stored in the endoplasmic reticulum or the Golgi apparatus. The multi-elemental analysis revealed that cobalt exposure significantly decreased magnesium and zinc content, with a likely competition of cobalt for magnesium and zinc binding sites in proteins. Overall, these data suggest a multiform toxicity of cobalt related to interactions with genomic DNA and nuclear proteins, and to the alteration of zinc and magnesium homeostasis.

Kinetic changes of oxidative stress and selenium status in plasma and tissues following burn injury in selenium-deficient and selenium-supplemented rats.

BACKGROUND: This study investigates the relationship between the burn-induced oxidative stress and the selenium status. METHODS: The rats were fed with a selenium-adequate diet or a selenium-depleted diet for 5 weeks, before a third-degree thermal injury was applied to the animals. One group of selenium-depleted animals received injections of sodium selenite after the injury. The selenium status and the oxidative stress parameters were measured for 5 days. RESULTS: The selenium-deficient diet leads to oxidative stress with a high stimulation of the superoxide dismutase activity. After the burn injury, the oxidative stress appears important because the initial selenium status is already impaired and, in all animals, the selenium levels and the antioxidant seleno-dependent glutathione peroxidase (GPx) activity decrease in the plasma and the tissues. A treatment with daily selenium injections is efficient in normalizing selenium levels and restores the GPx activity, but fails to counteract the initial oxidative damages induced by the selenium-deficient diet. CONCLUSIONS: The selenium status before the burn injury is a modulating factor of the burn-induced oxidative stress. A single selenium supplement is not sufficient to counteract these oxidative damages and henceforth combined antioxidant supplementations should be investigated to improve the early treatment of the burn patients.

Burn-induced alterations of chromium and the glucose/insulin system in rats.

Our objective was to demonstrate a role of chromium (Cr) in response to severe burn. A third-degree burn involving 20% of total body surface was applied under anaesthesia in accord with ethical guidelines. Chromium concentrations in liver decreased progressively and were non-detectable on days 5 and 10 following injury. In quadriceps muscle, Cr concentrations increased 6h after injury and then declined significantly within the first day and remained at these levels the following 9 days. Urinary Cr losses were also increased. Changes in kidney, brain and serum Cr were not significant. Non-fasting glucose rose 6h after injury and then returned to levels measured before the burn. There was a significant rise in corticosterone reaching a maximum the first day after injury that was accompanied by significant increases in circulating insulin and glucagon that were maximal after 2 days. Changes in IGF-1 were not significant. In summary, changes in Cr concentrations were associated with an early hyperglycemia, hyperinsulinemia and increased secretion of stress hormones. These observations strongly suggest a mobilization and utilization of Cr following severe burn. Additional studies are needed to document that improved Cr status might lead to improved recovery following burn.

Optimization of selenium status by a single intraperitoneal injection of Se in Se-deficient rat: possible application to burned patient treatment.

In order to investigate the efficiency of a single selenium (Se) administration in restoring selenium status, Se and antioxidant enzymes were studied in an animal model of Se depletion. In Se-depleted animals receiving or not a single parenteral administration of Se, plasma, red blood cell (RBC), and tissue Se levels were measured concurrently with glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities. The oxidative stress was assessed by thiobarbituric acid-reactive species (TBARs), total thiol groups, glutathione, and tocopherol measurements. Our study showed that Se depletion with alterations in the antioxidant defense system (Se and GPx activity decreases) led to an increase of lipid peroxidation, a decrease of the plasma vitamin E level, and SOD activation. Sodium selenite injection resulted after 24 h in an optimal plasma Se level and a reactivation of GPx activity. In liver, brain, and kidney, Se levels in injected animals were higher than those in reference animals. However, this single administration of Se failed to decrease free radical damage induced by Se depletion. Therefore, in burned patients who exhibit an altered Se status despite a daily usually restricted Se supplementation, the early administration of a consistent Se amount to improve the GPx activity should be of great interest in preventing the impairment of the antioxidant status.

Early evolution of selenium status and oxidative stress parameters in rat models of thermal injury.

The objective of the present study was to measure the relationship between selenium status and oxidative stress in two rat models of thermal injury. A non-lethal third-degree burn injury involving 20% (experiment 1) or 40% (experiment 2) of total body surface area (TBSA) was applied to male Wistar rats. Selenium level, glutathione peroxidase (GPx) activity in plasma, red blood cells (RBC) and tissues (liver, kidney, muscle, and brain), and plasma selenoalbumin (Se-alb) were measured in control rats and in burned rats respectively 6 hours after injury and daily from day 1 to day 5. In parallel, lipid and protein oxidative damages, monitored by plasma and tissue thiobarbituric acid reactive species (TBARs) levels and plasma total thiol groups were assessed. We observed a decrease of plasma Se and Se-albumin 6 hours after burn injury. In parallel, plasma GPx activity rapidly decreased and remained significantly lower than in control rats. These alterations were enhanced by the burn injury severity. Plasma TBARs followed the same pattern as that of plasma cholesterol, with an initial decrease and an increase at day 3 in 40% TBSA burned rats. Plasma thiol groups decreased in the two experiments indicating plasma protein oxidation. These results confirm an early oxidative stress in burn injury, and suggest an early selenium mobilization, which might counteract this oxidative stress. These data underline the crucial need of a restored selenium status in burned patients immediately after the burn injury.