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  • Title: Recovery potential of hepatocytes from inhibition of albumin secretion by cadmium.
    Author: Wan X, Lachapelle M, Marion M, Fournier M, Denizeau F.
    Journal: J Toxicol Environ Health; 1993 Apr; 38(4):381-92. PubMed ID: 8478980.
    Abstract:
    The aim of this study was to examine albumin production, a typical liver-specific function, in hepatocytes treated with Cd and to examine the reversibility of the perturbations induced by the toxic metal. Cultures of freshly isolated rat hepatocytes were exposed to increasing amounts of Cd in modified Leibowitz L-15 medium for 20 h; the cells were then allowed to recover by further incubation in Cd-free medium for an additional period of 20 h. The levels of albumin secreted into the extracellular medium were determined by enzyme-linked immunosorbent assay and were found to be reduced by Cd in a concentration-dependent fashion over the first 20 h. Inhibition was seen at Cd concentrations that did not cause any loss of cellular viability (up to 0.5 microM Cd), as judged from the release of lactate dehydrogenase by the cells. After replacement of the exposure medium by Cd-free medium, the same pattern of diminished albumin secretion was obtained, revealing the persistence of the cytotoxic effects when recovery conditions were applied. Moreover, hepatocytes exposed to 0.5 microM Cd for 20 h and processed for visualization of albumin immunoreactive sites using protein A-gold and electron microscopy exhibited very low albumin-specific labeling as compared to the controls (0.6 +/- 0.05 vs. 20.0 +/- 2.6 gold particles/micron2). Intracellular glutathione levels were not significantly changed by Cd either after the initial exposure or after the incubation that followed in control medium. The accumulation of Cd by the cells, as measured by graphite furnace atomic absorption spectrophotometry, was concentration dependent. It remained stable after medium change, indicating that Cd efflux was negligible upon reestablishment of normal conditions. The present data show that the perturbations in albumin metabolism caused by Cd are not readily alleviated after the cells are returned to Cd-free medium, suggesting a limited short-term recovery potential against cytotoxic damage. The data also demonstrate that hepatocyte-specific functions can be used as sensitive indicators for the detection of cellular disturbances by hepatotoxins.
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