Glucuronidation of 7-hydroxycoumarin in periportal and pericentral regions of the lobule in livers from untreated and 3-methylcholanthrene- treated rats

JG Conway, FC Kauffman, T Tsukuda and RG Thurman

Department of Pharmacology, University of North Carolina, Chapel Hill 27514.

Rates of production of 7-hydroxycoumarin glucuronide were measured in specific zones of the liver lobule using micro-light guides placed on periportal and pericentral regions on the surface of livers from untreated and 3-methylcholanthrene-treated rats. Livers were perfused with sulfate-free buffer under normoxic conditions and fluorescence of free 7-hydroxycoumarin was monitored. The formation of nonfluorescent 7- hydroxycoumarin glucuronide was then inhibited completely by perfusion with N2-saturated perfusate containing 20 mM ethanol. The difference between fluorescence readings under normoxic and hypoxic conditions was used to calculate rates of glucuronidation. Maximal rates of glucuronidation (11.9-13.5 mumol/g/hr) did not differ significantly in periportal and pericentral regions in livers from either 3- methylcholanthrene-treated or untreated rats. In all regions of the liver lobule, glucuronidation was half-maximal with about 20 microM 7- hydroxycoumarin. Glucuronosyltransferase assayed in lyophilized tissue sections with saturating concentrations of UDPGA (9 mM) was 2.3-fold greater in pericentral than in periportal areas in livers from untreated rats. In livers from 3-methylcholanthrene-treated rats, activities were similar in periportal and pericentral regions but were 4- to 7-fold higher than values from untreated rats. In addition, glucuronosyltransferase activity assayed in native microsomes with physiological concentrations of UDP-glucuronic acid (UDPGA) (0.4 mM) with UDP-N-acetylglucosamine (0.3 mM) was 2-fold higher in preparations from 3-methylcholanthrene-treated than untreated rats. Thus, 3- methylcholanthrene treatment increased glucuronosyltransferase activity in vitro but did not alter rates of glucuronide formation in periportal and pericentral regions of the liver lobule of intact liver. Infusion of epinephrine (50 nM) into perfused livers from untreated and 3- methylcholanthrene-treated rats increased rates of glucuronidation by about 35%. Since epinephrine probably acts by increasing the supply of the cofactor UDPGA due to increased breakdown of glycogen, it follows that UDPGA supply limits rates of glucuronidation in perfused livers from both untreated and 3-methylcholanthrene-treated rats.

Volume 33, Issue 1, pp. 111-119, 01/01/1988
Copyright © 1988 by American Society for Pharmacology and Experimental Therapeutics

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