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Rifampicin Induces Bicarbonate-Rich Choleresis in Rats: Involvement of Anion Exchanger 2

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Abstract

Background and Aim

Previous studies have shown that rifampicin induced choleresis, the mechanisms of which have not been described. The aim of this study was to investigate the mechanisms underlying in vivo rifampicin-induced choleresis.

Methods

In one experimental set, rats were treated chronically with rifampicin on days 1, 3 and 7. Serum and biliary parameters were assayed, and mRNA and protein levels, as well as the locations of the hepatic export transporters were analyzed by real-time PCR, western blot and immunofluorescence. Ductular mass was evaluated immunohistochemically. In another experimental set, rats received an acute infusion of rifampicin. The amount of rifampicin in bile was detected using HPLC. Biliary parameters were monitored following intrabiliary retrograde fluxes of the Cl−/HCO3 − exchange inhibitor 4,4′-diisothiocyanatostilbene-2,2′-disulfonic acid (DIDS) or 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB) in the infused rats.

Results

Biliary bicarbonate output increased in parallel to the augmented bile flow in response to rifampicin, and this effect was abolished with intrabiliary administration of DIDS, but not NPPB. The biliary secretion of rifampicin with increases in bile flow and biliary rifampicin in response to different infused doses of the antibiotic show no significant correlations. After rifampicin treatment, the expression level of anion exchanger 2 (AE2) increased, while the location of hepatic transporters did not change. However, RIF treatment did not increase ductular mass significantly.

Conclusions

These results indicate that the increase in bile flow induced by rifampicin is mainly due to increased HCO3 − excretion mediated by increased AE2 protein expression and activity.

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Abbreviations

RIF:

Rifampicin

ALT:

Alanine aminotransferase

TBA:

Total bile acids

GSH:

Reduced glutathione

Bsep:

Bile salt export pump

Mrp2:

Multidrug resistance-associated protein 2

AE2:

Anion exchanger 2

Mrp4:

Multidrug resistance-associated protein 4

BSDF:

Bile salt-dependent component of bile flow

BSIF:

Bile salt-independent fraction of bile flow

DMSO:

Dimethylsulfoxide

RT-PCR:

Real-time polymerase chain reaction

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

HPLC:

High-performance liquid chromatography

PBC:

Primary biliary cirrhosis

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Acknowledgments

This study was supported by grants from the Natural Science Foundation of China (Grant No. 81370529).

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Authors and Affiliations

  1. Department of Gastroenterology, The First Affiliated Hospital, Anhui Medical University, Hefei, 230022, Anhui Province, China

    Wei Wang, Xiaofei Ren, Yi Cai, Lihong Chen, Weiping Zhang & Jianming Xu

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  1. Wei Wang
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Correspondence to Jianming Xu.

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Research involving human participants and/or animals

All animals received humane care, and all testing methods were performed in accordance with the Institutional Guidelines of the Anhui Medical University for the Care and Use of Laboratory Animals; the study methods were carried out under the protocol approved by the Institutional Animal Care and Use Committee of the Anhui Medical University. This article does not contain any studies with human participants performed by any of the authors.

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Wang, W., Ren, X., Cai, Y. et al. Rifampicin Induces Bicarbonate-Rich Choleresis in Rats: Involvement of Anion Exchanger 2. Dig Dis Sci 61, 126–136 (2016). https://doi.org/10.1007/s10620-015-3850-2

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  • DOI: https://doi.org/10.1007/s10620-015-3850-2

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