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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

Comparative cytopathology of primary rat hepatocyte cultures exposed to aflatoxin B1, acetaminophen, and other hepatotoxins.

The cytomorphological alterations in primary monolayer cultures of Fischer 344 rat hepatocytes exposed continuously to lethal concentrations of various hepatotoxins were compared. Aflatoxin B1 (AFB1) at concentrations of 1 and 10 microM killed 67 and 97% hepatocytes, respectively, at 48 hr, as determined by release of lactate dehydrogenase (LDH) into culture medium, or by trypan blue uptake. AFB1 produced numerous radial finger-like projections (blebs) at attached margins of cytoplasm in almost all hepatocytes between 6 and 24 hr. Formation of blebs was dose dependent and preceded release of LDH and trypan blue uptake. Similar marginal blebs were produced by cycloheximide, 2-acetylaminofluorene, N-hydroxy-2-acetylaminofluorene, and senecionine. By comparison, acetaminophen, at equivalently lethal concentrations (4 or 16 mM) did not cause marginal blebs, but resulted in a similar time course of LDH release and trypan blue uptake. Carbon tetrachloride and bromobenzene also failed to produce blebs at lethal concentrations. Phalloidin and cytochalasin B rapidly produced smaller spherical blebs over the entire surface of all hepatocytes, distinct from the finger-like blebs produced only at the free attached margins of cells exposed to AFB1. Blebs and lethal injury by AFB1 were reduced by phenobarbitone or 3-methylcholanthrene pretreatments and by SKF-525-A in culture. The demonstration of morphological prelethal changes in hepatocytes injured by different classes of hepatotoxins in culture provides a means of differentiating the early biochemical mechanisms by which hepatotoxins and hepatocarcinogens lethally injure hepatocytes before membrane permeability to LDH and trypan blue is detectable.[1]


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