Sinusoidal efflux of taurocholate correlates with the hepatic expression level of Mrp3

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Abstract

Multidrug resistance-associated protein 3 (Mrp3/ABCC3), which can mediate the cellular extrusion of bile acids, is induced on the hepatic sinusoidal membrane of Mrp2/ABCC2-deficient rats (Eisai hyperbilirubinemic rats; EHBRs) and phenobarbital-treated Sprague–Dawley rats. In the present study, the correlation between the sinusoidal efflux clearance (PSeff) of [3H]taurocholate (TC) and the hepatic expression of Mrp3 was investigated using perfused liver from these rats. A significant correlation was observed between the PSeff and the hepatic expression level of Mrp3, suggesting a contribution by Mrp3 to the sinusoidal efflux of TC. The results of the kinetic analysis also suggested that other transporter(s) on the sinusoidal plasma membrane may participate in the efflux of TC under physiological conditions. The contribution of Mrp3 to the sinusoidal efflux of TC in EHBRs and phenobarbital (80 and 40 mg/kg)-treated rats was revealed to be 58%, 48%, and 31%, respectively.

Section snippets

Materials and methods

Materials. Unlabeled and [3H]-labeled TCs (2.0 μCi/nmol) were purchased from Sigma Chemical (St. Louis, MO) and New England Nuclear (Boston, MA), respectively. [14C]-Labeled inulin (2.64 mCi/g) was purchased from New England Nuclear. PB was purchased from Sigma Chemical. Other chemicals used were commercially available and reagent grade products. Male SD rats and EHBRs (7–8 weeks old) were purchased from Nihon SLC (Shizuoka, Japan). Rats were kept in an SPF room until they were 11 weeks old and

Disposition of [3H]TC in the perfused liver of PB-treated SD rats and control SD rats

Previously, we compared the hepatic disposition of [3H]TC in SD rats and EHBRs [37]. In the present study, the isolated livers from control and PB-treated SD rats were additionally perfused with the Krebs–Ringer bicarbonate buffer containing 0.1 μM [3H]TC. The time-profiles for excretion into bile and outflow are shown in Figs. 1A and B, respectively. At 40 min, the value of Vbile/I in high dose PB-treated rats (0.77 ± 0.04) was significantly (P<.05) lower than that in low dose PB-treated (0.89 ± 

Discussion

We have previously demonstrated that the PSeff of [3H]TC from hepatocytes to blood is enhanced in EHBRs whose Mrp3 expression level is induced [37], and suggested that Mrp3 may participate in the sinusoidal efflux of [3H]TC. To confirm this hypothesis, in the present study, the PSeff values were also compared in control and PB-treated rats, whose expression level of Mrp3 was enhanced. Kinetic analysis of the washout experiments showed that the PSnet,eff in high dose PB-treated rats was three

Acknowledgements

This work was supported by Grants-in-Aid for Scientific Research on Priority Areas on ABC Proteins 10044243 and on Priority Areas on Epithelial Vectorial Transport 12144201 from the Ministry of Education, Science, and Culture of Japan.

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