A Study of Bile Canalicular Contractions in Isolated Hepatocytes

Phillips, M. James; Oshio, Chikara; Miyairi, Mamoru; Katz, Hart; Smith, Charles R.

doi:10.1002/hep.1840020603

Cited by 70 publications

(35 citation statements)

References 12 publications

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“…1; Table 4). Physiological mechanisms may explain this flux; actin filamentmediated contractility of bile canaliculi facilitates bile flow in vivo in rat liver (Watanabe et al, 1991), and the regular, ordered contraction of bile canaliculi has been reported previously in vitro in isolated hepatocyte couplets/hepatocyte groups (Oshio and Phillips, 1981;Phillips et al, 1982).…”

Section: Discussionmentioning

confidence: 66%

Hepatobiliary Disposition of Troglitazone and Metabolites in Rat and Human Sandwich-Cultured Hepatocytes: Use of Monte Carlo Simulations to Assess the Impact of Changes in Biliary Excretion on Troglitazone Sulfate Accumulation

Lee¹,

Marion²,

Abe³

et al. 2009

J Pharmacol Exp Ther

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This study examined the hepatobiliary disposition of troglitazone (TGZ) and metabolites [TGZ sulfate (TS), TGZ glucuronide (TG), and TGZ quinone (TQ)] over time in rat and human sandwich-cultured hepatocytes (SCH). Cells were incubated with TGZ; samples were analyzed for TGZ and metabolites by liquid chromatography-tandem mass spectrometry. SCH mimicked the disposition of TGZ/metabolites in vivo in rats and humans; TGZ was metabolized primarily to TS and to a lesser extent to TG and TQ. In human SCH, the biliary excretion index (BEI) was negligible for TGZ and TQ, ϳ16% for TS, and ϳ43% for TG over the incubation period; in rat SCH, the BEI for TS and TG was ϳ13 and ϳ41%, respectively. Hepatocyte accumulation of TS was extensive, with intracellular concentrations ranging from 132 to 222 M in rat SCH; intracellular TGZ concentrations ranged from 7.22 to 47.7 M. In human SCH, intracellular TS and TGZ concentrations ranged from 136 to 160 M and from 49.4 to 84.7 M, respectively. Pharmacokinetic modeling and Monte Carlo simulations were used to evaluate the impact of modulating the biliary excretion rate constant (K bile ) for TS on TS accumulation in hepatocytes and medium. Simulations demonstrated that intracellular concentrations of TS may increase up to 3.1-and 5.7-fold when biliary excretion of TS was decreased 2-and 10-fold, respectively. It is important to note that altered hepatobiliary transport and the extent of hepatocyte exposure may not always be evident based on medium concentrations (analogous to systemic exposure in vivo). Pharmacokinetic modeling/simulation with data from SCH is a useful approach to examine the impact of altered hepatobiliary transport on hepatocyte accumulation of drug/metabolites.

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Section: Discussionmentioning

confidence: 66%

Hepatobiliary Disposition of Troglitazone and Metabolites in Rat and Human Sandwich-Cultured Hepatocytes: Use of Monte Carlo Simulations to Assess the Impact of Changes in Biliary Excretion on Troglitazone Sulfate Accumulation

Lee¹,

Marion²,

Abe³

et al. 2009

J Pharmacol Exp Ther

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show abstract

“…1) (Pfeifer et al, 2014). Regular, ordered contraction of bile canaliculi has been reported previously in isolated couplets and cultured hepatocytes (Oshio and Phillips, 1981;Phillips et al, 1982), and has been shown to facilitate bile flow in vivo in rat liver (Watanabe et al, 1991). In the study by Jemnitz et al (2010), basolateral efflux was evaluated by measuring TCA in standard buffer during the efflux phase.…”

Section: Discussionmentioning

confidence: 94%

Species Differences in Hepatobiliary Disposition of Taurocholic Acid in Human and Rat Sandwich-Cultured Hepatocytes: Implications for Drug-Induced Liver Injury

Yang

Pfeifer

Köck

et al. 2015

J Pharmacol Exp Ther

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The bile salt export pump (BSEP) plays an important role in bile acid excretion. Impaired BSEP function may result in liver injury. Bile acids also undergo basolateral efflux, but the relative contributions of biliary (CL Bile ) versus basolateral efflux (CL BL ) clearance to hepatocellular bile acid excretion have not been determined. In the present study, taurocholic acid (TCA; a model bile acid) disposition was characterized in human and rat sandwich-cultured hepatocytes (SCH) combined with pharmacokinetic modeling. In human SCH, biliary excretion of TCA predominated (CL Bile 5 0.14 6 0.04 ml/min per g liver; CL BL 5 0.042 6 0.019 ml/min per g liver), whereas CL Bile and CL BL contributed approximately equally to TCA hepatocellular excretion in rat SCH (CL Bile 5 0.34 6 0.07 ml/min per g liver; CL BL 5 0.26 6 0.07 ml/min per g liver).Troglitazone decreased TCA uptake, CL Bile , and CL BL ; membrane vesicle assays revealed for the first time that the major metabolite, troglitazone sulfate, was a noncompetitive inhibitor of multidrug resistance-associated protein 4, a basolateral bile acid efflux transporter. Simulations revealed that decreased CL Bile led to a greater increase in hepatic TCA exposure in human than in rat SCH. A decrease in both excretory pathways (CL Bile and CL BL ) exponentially increased hepatic TCA in both species, suggesting that 1) drugs that inhibit both pathways may have a greater risk for hepatotoxicity, and 2) impaired function of an alternate excretory pathway may predispose patients to hepatotoxicity when drugs that inhibit one pathway are administered. Simulations confirmed the protective role of uptake inhibition, suggesting that a drug's inhibitory effects on bile acid uptake also should be considered when evaluating hepatotoxic potential. Overall, the current study precisely characterized basolateral efflux of TCA, revealed species differences in hepatocellular TCA efflux pathways, and provided insights about altered hepatic bile acid exposure when multiple transport pathways are impaired.

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“…From this morphological analogy in the filament distribution, it is interesting to speculate that bile canaliculi are capable of active contraction like an intestinal epithelium. Recently, Oshio and Phillips (1981), and Phillips et al (1982Phillips et al ( , 1983 showed that cultured rat hepatocytes really had a forceful contraction ability. As yet, there has been no clear evidence that bile canaliculi in human do contract in vivo, and further study must be performed to substantiate this speculation.…”

Section: Discussionmentioning

confidence: 99%

Three-dimensional structure of cytoskeletal system in human hepatocytes viewed by polyethylene-glycol-embedding method.

Ishii

Miyazaki

Ohta

et al. 1985

Tohoku J. Exp. Med.

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In an attempt to demonstrate the three-dimensional cytoskeletal system of human hepaocytes, we used two kinds of tissue processing. In one, specimens were immediately fixed with 1% glutaraldehyde, saponin 0.5 mg/ml, and tannic acid 2 mg/ml (procedure A) and in the other, cytoplasmic matrix and organelles were washed out with a solution containing saponin 0.5 mg/ml and 0.15% Triton X-100, followed by fixation with glutaraldehyde (procedure B). The specimens were embedded in polyethylene glycol, sectioned at 500 nm, critical point-dried and observed in a TEM. With procedure A, a fine lattice was shown to spread throughout the cytoplasm so as to connect plasma membranes with cytoplasmic organelles, condensed around the bile canaliculi. With procedure B, microfilaments, microtubules and intermediate filaments could be clearly visualized three-dimensionally instead of the lattice structure. At a higher magnification, microvilli of canaliculus were found to contain a core microfilament which were enmeshed in a pericanalicular filament plexus. This plexus looking like the terminal web of intestinal epithelia, encircled the canaliculus and was attached to the zonula adherens. Outside the plexus, intermediate filament network was shown attached to the desmosome. These morphological findings indicated that the cytoskeleton of human hepatocytes is composed of microfilaments, microtubules, intermediate filaments and the fine lattice structure, suggesting that the bile canaliculi could contract in vivo.cytoskeleton ; human hepatocyte ; polyethylene-glycol-embedding method ; three-dimensional electron microscopy Many papers have dealt with the filamentous structures of human hepatocytes and their relation to bile canaliculi (Bieva 1964;Oda et al. 1974;French and Davis 1975;Yokota and Fahimi 1979). The distribution of the filaments can be observed by using immunofluorescence staining (Horoboroco et al. 1975;Yokota and Fahimi 1979), but can hardly be seen by conventional electron microscopy because in a two dimensional ultrathin section filamentous structures appear only

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A Study of Bile Canalicular Contractions in Isolated Hepatocytes

Cited by 70 publications

References 12 publications

Hepatobiliary Disposition of Troglitazone and Metabolites in Rat and Human Sandwich-Cultured Hepatocytes: Use of Monte Carlo Simulations to Assess the Impact of Changes in Biliary Excretion on Troglitazone Sulfate Accumulation

Hepatobiliary Disposition of Troglitazone and Metabolites in Rat and Human Sandwich-Cultured Hepatocytes: Use of Monte Carlo Simulations to Assess the Impact of Changes in Biliary Excretion on Troglitazone Sulfate Accumulation

Species Differences in Hepatobiliary Disposition of Taurocholic Acid in Human and Rat Sandwich-Cultured Hepatocytes: Implications for Drug-Induced Liver Injury

Three-dimensional structure of cytoskeletal system in human hepatocytes viewed by polyethylene-glycol-embedding method.

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