Cytotoxicity of ethanol in primary cultures of rat midbrain neurons.

Primary cultures of midbrain neurons were obtained from 15-day-old rat fetuses. Neuron cultures were exposed to ethanol (27 mM, 43 mM and 120 mM) for 24 h and evaluated by light microscopy, a viability measure, and protein content. Ethanol concentrations of 43 and 120 mM appeared to affect the cultures both in terms of cell viability and protein. This effect was independent of any osmotic effect, when sucrose was run as a control. We conclude that primary cultures of midbrain neurons are sensitive to relatively low concentrations of ethanol, compared to cell culture preparations used by other investigators.

Acetaldehyde-induced lipid peroxidation in isolated hepatocytes.

In an effort to evaluate further the concept of ethanol-induced lipid peroxidation, isolated rat hepatocytes obtained via collagenase perfusion were utilized. Hepatocytes were judged to be functionally intact based on measurements of adenosine-5-triphosphate, gluconeogenesis, bromosulphthalein uptake, and trypan blue exclusion. When hepatocytes were incubated with acetaldehyde, a metabolite of ethanol, at 100 mg% and 10 mg%, significant increases in lipid peroxidation resulted as measured by levels of malonaldehyde. Acetaldehyde-induced increases in malonaldehyde were reduced by pre-incubation with antioxidants such as Vitamin E (200 mg%) or glutathione (100 mg%).

Levels of lipid peroxidation in hepatocytes isolated from aging rats fed an antioxidant-free diet.

Enzymatically isolated hepatocytes were utilized to evaluate levels of lipid peroxidation in young (3 months), adult (12 months) and aged (25 months) Fisher-344 female rats. Lipid peroxidation was measured by assaying levels of malonaldehyde, a by-product of the peroxidation reaction. Young, adult and aged animals were fed a liquid antioxidant-free diet for 21 days prior to the hepatocyte isolation. Cells isolated from young rats demonstrated the highest levels of lipid peroxidation (microgram of malonaldehyde/10(6) cells). This increase may be a result of the isolation procedure or of metabolic differences found in younger animals. When hepatocytes from aged animals on the antioxidant-free diet were incubated in the presence of lipid peroxidation inducers, such as cumene hydroperoxide or NADPH, significant elevations in lipid peroxidation over comparable adult values were observed. Aged hepatocytes appear to be more susceptible to the peroxidation process than adult hepatocytes, possibly due to defective peroxidation defenses.

Influence of chronic ethanol vapor inhalation on hepatic parenchymal and Kupffer cell function.

This report evaluates the influence of chronic ethanol vapor inhalation on various facets of liver function as well as induction of hepatic and pulmonary pathology in rats. Chronic ethanol inhalation is associated with minimal hepatic dysfunction, but pronounced induction of metabolic tolerance.

Comparative uptake of sulfobromophthalein by isolated Kupffer and parenchymal cells.

The relative role of specific liver cells in the uptake of sulfobromophthalein (BSP) was ascertained by utilizing enzymatically isolated rat hepatic Kupffer and parenchymal cells. Kupffer cells demonstrated the ability neither to remove BSP from the incubation medium nor to form a BSP-glutathione conjugate. In contrast, parenchymal cells removed BSP from the medium and formed a BSP-glutathione conjugate. The rate and maximum uptake of BSP by the parenchymal cells were inversely related to the concentration of serum or albumin in the incubation medium. In an effort to evaluate the influence of ethanol on BSP uptake, parenchymal cells were incubated in the presence of varying concentrations of ethanol. No alteration in BSP uptake was induced by the prior addition of ethanol to the incubation medium. The uptake and conjugation of BSP are exclusive functional expressions of the hepatic parenchymal cell population.