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  • Title: Trypan blue uptake as a new method to investigate hepatotoxicity in periportal and pericentral regions of the liver lobule: studies with allyl alcohol in the perfused liver.
    Author: Belinsky SA, Popp JA, Kauffman FC, Thurman RG.
    Journal: J Pharmacol Exp Ther; 1984 Sep; 230(3):755-60. PubMed ID: 6470978.
    Abstract:
    A new method based on trypan blue uptake was developed to study zonal hepatotoxicity in specific regions of the liver lobule in perfused livers. Allyl alcohol (350 microM), a hepatotoxin that causes necrosis of periportal regions, was infused into livers from phenobarbital-treated rats for 60 min. The subsequent infusion of trypan blue (0.2 mM) stained nuclei in periportal but not midzonal or pericentral regions of the liver lobule. With this new method of dye uptake, comparison of the temporal relationship between metabolic changes and the onset of cellular damage was possible. During the first 10 min of allyl alcohol infusion, allyl alcohol was taken up at rates of 64 mumol/g/h concomitant with an increase in oxygen uptake of 18 mumol/g/h. Allyl alcohol infusion also increased bile flow from 69 to 167 microliters/min and release of oxidized glutathione from 127 to 217 nmol/g/h, respectively. However, during the next 20-min phase of exposure to allyl alcohol, oxygen uptake was inhibited by 60% and allyl alcohol uptake declined by 80%. Bile flow and release of oxidized glutathione also declined continuously between 10 and 30 min and by 30 min stopped completely. Concomitant with the inhibition of oxygen uptake, rates of glycogenolysis and glycolysis increased by 130 and 85%, respectively. Within minutes after maximal inhibition of oxygen uptake, between 30 and 40 min after initiation of allyl alcohol infusion, trypan blue was taken up by hepatocytes in periportal zones of the liver lobule. Subsequently, lactate dehydrogenase began to appear in the effluent perfusate.(ABSTRACT TRUNCATED AT 250 WORDS)
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