Elsevier

Toxicology

Volume 121, Issue 2, 15 August 1997, Pages 127-142
Toxicology

Role of bile salts in colchicine-induced hepatotoxicity. Implications for hepatocellular integrity and function1

https://doi.org/10.1016/S0300-483X(97)00064-4Get rights and content

Abstract

Colchicine, a microtubule-disrupting agent, induces hepatotoxicity in experimental animals at the doses commonly employed to explore vesicular transport in the liver. The effect of manipulations of the bile salt pool on colchicine-induced hepatotoxicity was studied in rats to determine the role of bile salts in this phenomenon. Leakage of enzyme markers of liver-cell damage into plasma and bile induced by colchicine pre-treatment displayed a sigmoidal log dose-effect curve, the half-maximal effect being reached at 0.12 μmol per 100 g body wt. Lumicolchicine, instead, showed no harmful effect. Maximal increment of biliary LDH discharge induced by colchicine was reduced from 950±124% to 216±29% by bile diversion leading to a marked reduction in bile salt output, and this parameter was further decreased to 100±13% and 157±39% by subsequent repletion of the bile salt pool with the hydrophilic bile salts taurodehydrocholate and tauroursodeoxycholate, respectively. Conversely, infusion of taurocholate into non-bile salt depleted, colchicine-treated rats led to cholestasis and massive discharge of enzymes into both blood and bile. Our data show conclusively that colchicine-induced hepatotoxicity depends on the magnitude and composition of the bile salt flux traversing the liver. They also support the view that functional integrity of vesicular mechanisms presumably involved in membrane repair are indispensable to protect the hepatocytes from the damaging effect of bile salts during normal bile formation.

Introduction

Colchicine, a microtubule-disrupting agent (Borisy and Taylor, 1967; Wilson, 1975), is used as a therapeutic agent for a number of diseases including gout, amyloidosis, cirrhosis, and familial Mediterranean Fever, among others (Levy et al., 1991). In addition, this drug is being extensively employed as a tool to obtain experimental evidence of vesicular, microtubule-dependent transport mechanisms in the liver. Indeed, colchicine-induced reduction of the release into blood and/or bile of several compounds including proteins (Barnwell and Coleman, 1983; Godfrey et al., 1982; Mullock et al., 1980; Reaven and Reaven, 1980; Redman et al., 1975; Stein and Stein, 1973; Stein et al., 1974), cholesterol and phospholipids (Barnwell, et al., 1984; Crawford et al., 1988; Gregory et al., 1978), bile salts (Crawford et al., 1988; Crawford et al., 1993; Dubin et al., 1980; Erlinger, 1996; Fukumoto et al., 1982), inert solutes (LeSage et al., 1990; Roma et al., 1989; Scharschmidt et al., 1986) and cholephilic dyes (Aoyama et al., 1991; Aoyama et al., 1991b; Mori et al., 1987) has been considered indicative of transcytotic pathways for the transport of these compounds in the liver.

In addition to causing vesicular arrest, colchicine has been shown to induce conspicuous hepatotoxic effects in experimental animals, including fatty liver and necrosis (Stein and Stein, 1973) as well as elevation of serum transaminase levels (Dubin et al., 1980; Fukumoto et al., 1982; Rao and Mehendale, 1991), when administered at the doses commonly used to produce a marked reduction of the number of microtubules. To elucidate the factors determining such harmful effect is of interest, however, not only to establish the relative contribution of this alteration to the impaired transport of compounds thought to reach bile via a vesicular mechanism but also to better understand the role of microtubules in the preservation of hepatocyte integrity.

Colchicine induces cholestasis and impairs bile salt excretion when micelle-forming, but not non-micelle forming bile salts are administered at doses well below those reported to induce hepatocyte injury in non-treated animals (Barnwell et al., 1984; Crawford et al., 1988; Dubin et al., 1980, Nakai et al., 1992). Based on this previous finding, it could be hypothesized that the hepatotoxic effects induced by colchicine under basal conditions of bile salt excretion is the consequence of the high susceptibility of these animals to the cholestatic effect induced by bile salts. To corroborate this assumption, we have studied in detail the role of bile salts in colchicine-induced hepatotoxicity. For this purpose, a formal dose-response study was carried out by monitoring changes in the release of enzyme markers of liver-cell damage following colchicine administration under basal conditions of bile secretion. Next, the influence of different manipulations of the bile salt pool was assessed, including bile salt administration, bile salt depletion, and bile salt depletion followed by repletion of the bile salt pool with hydrophilic, non-hepatotoxic bile salts. Our results indicate that colchicine-induced hepatotoxicity depends on the magnitude and composition of the pool of bile salts traversing the liver, thus pointing to a role of bile salts in colchicine-induced liver-cell damage.

Section snippets

Chemicals

Sodium taurocholate (TC), 3α-hydroxy-steroid dehydrogenase and colchicine were obtained from Sigma (St. Louis, MO). Sodium taurodehydrocholate (TDHC) was purchased from Calbiochem (La Jolla, CA). Sodium tauroursodeoxycholate (TUDC) was a generous gift from Prodotti Chimici e Alimentari S.p.A. (Genova, Italy). Bile salts were all more than 98% pure when examined by HPLC. Lumicolchicine was prepared by exposure of colchicine to ultraviolet light as indicated previously (Dubin et al., 1980), and

Dose-effect study

As shown in Fig. 2, a typical sigmoidal dose-effect curve was obtained when the magnitude of the changes in biliary LDH output induced by colchicine administration (expressed as the percent of the increase in this parameter with respect to the basal values) were plotted against the logarithm of the dose administered of colchicine. An apparent maximal effect was reached at the dose of 0.5 μmol per 100 g body wt, which amounted to an increment of 950%. The dose at which colchicine was

Discussion

The elucidation of the mechanisms responsible for the harmful effect of colchicine is of interest to better understand the role of the vesicular trafficking to the canalicular membrane in the preservation of the structural and functional integrity of the hepatocyte apical pole. In fact, a body of published data showed that microtubule disruption may interfere not only with the delivery of compounds destined to bile secretion but also with the normal supply of newly synthesized constitutive

Acknowledgements

This work was supported financially by a Research Grant from Consejo Nacional de Investigaciones Cientı́ficas y Técnicas (CONICET), Argentina. The authors wish to thank Proffesor Roger Coleman for helpful comments and suggestions in reviewing the manuscript.

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