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Differential Contributions of rOat1 (Slc22a6) and rOat3 (Slc22a8) to the in Vivo Renal Uptake of Uremic Toxins in Rats

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Purpose.

Evidence suggests that uremic toxins such as hippurate (HA), indoleacetate (IA), indoxyl sulfate (IS), and 3-carboxy-4-methyl-5-propyl-2-furanpropionate (CMPF) promote the progression of renal failure by damaging tubular cells via rat organic anion transporter 1 (rOat1) and rOat3 on the basolateral membrane of the proximal tubules. The purpose of the current study is to evaluate the in vivo transport mechanism responsible for their renal uptake.

Methods.

We investigated the uremic toxins transport mechanism using the abdominal aorta injection technique [i.e., kidney uptake index (KUI) method], assuming minimal mixing of the bolus with serum protein from circulating serum.

Results.

Maximum mixing was estimated to be 5.8% of rat serum by measuring estrone sulfate extraction after addition of 0–90% rat serum to the arterial injection solution. Saturable renal uptake of p-aminohippurate (PAH, Km = 408 μM) and benzylpenicillin (PCG, Km = 346 μM) was observed, respectively. The uptake of PAH and PCG was inhibited in a dose-dependent manner by unlabeled PCG (IC50 = 47.3 mM) and PAH (IC50 = 512 μM), respectively, suggesting that different transporters are responsible for their uptake. A number of uremic toxins inhibited the renal uptake of PAH and PCG. Excess PAH, which could inhibit rOat1 and rOat3, completely inhibited the saturable uptake of IA, IS, and CMPF by the kidney, and by 85% for HA uptake. PCG inhibited the total saturable uptake of HA, IA, IS, and CMPF by 10%, 10%, 45%, and 65%, respectively, at the concentration selective for rOat3.

Conclusions.

rOat1 could be the primary mediator of the renal uptake of HA and IA, accounting for approximately 75% and 90% of their transport, respectively. rOat1 and rOat3 contributed equally to the renal uptake of IS. rOat3 could account for about 65% of the uptake of CMPF under in vivo physiologic conditions. These results suggest that rOat1 and rOat3 play an important role in the renal uptake of uremic toxins and the induction of their nephrotoxicity.

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Abbreviations

CMPF:

3-carboxy-4-methyl-5-propyl-2-furanpropionate

ES:

estrone sulfate

HA:

hippurate

IA:

indoleacetate

IS:

indoxyl sulfate

KUI:

kidney uptake index

PAH:

p-aminohippurate

PCG:

benzylpenicillin; rOat, rat organic anion transporter

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Authors and Affiliations

  1. Department of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto, 862-0973, Japan

    Tsuneo Deguchi, Yousuke Kouno & Masaki Otagiri

  2. Department of Molecular Biopharmacy and Genetics, Graduate School of Pharmaceutical Sciences, Tohoku University, Aoba, Aramaki, Aoba-ku, Sendai, 980-8578, Japan

    Tetsuya Terasaki

  3. New Industry Creation Hatchery Center, Tohoku University, Aoba, Aramaki, Aoba-ku, Sendai, 980-8578, Japan

    Tetsuya Terasaki

  4. Solution-Oriented Research for Science and Technology of Japan Science and Technology Agency, Saitama, 332-0012, Japan

    Tetsuya Terasaki

  5. Daiichi College of Pharmaceutical Sciences, Minami-ku, Fukuoka, 815-0037, Japan

    Akira Takadate

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  1. Tsuneo Deguchi
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  2. Yousuke Kouno
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  3. Tetsuya Terasaki
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  4. Akira Takadate
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  5. Masaki Otagiri
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Correspondence to Masaki Otagiri.

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Deguchi, T., Kouno, Y., Terasaki, T. et al. Differential Contributions of rOat1 (Slc22a6) and rOat3 (Slc22a8) to the in Vivo Renal Uptake of Uremic Toxins in Rats. Pharm Res 22, 619–627 (2005). https://doi.org/10.1007/s11095-005-2486-x

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  • DOI: https://doi.org/10.1007/s11095-005-2486-x

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