Differential contributions of rOat1 (Slc22a6) and rOat3 (Slc22a8) to the in vivo renal uptake of uremic toxins in rats.
OBJECTIVE
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, K(m) = 408 microM) and benzylpenicillin (PCG, K(m) = 346 microM) was observed, respectively. The uptake of PAH and PCG was inhibited in a dose-dependent manner by unlabeled PCG (IC(50) = 47.3 mM) and PAH (IC(50) = 512 microM), 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.
Deguchi
Tsuneo
T
Department of Biopharmaceutics, Graduate School of Pharmaceutical Sciences, Kumamoto University, Kumamoto 862-0973, Japan.
Kouno
Yousuke
Y
Terasaki
Tetsuya
T
Takadate
Akira
A
Otagiri
Masaki
M
D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002250:Carbon Radioisotopes; D004970:Estrone / Q000031:analogs & derivatives / Q000378:metabolism; D005663:Furans / Q000493:pharmacokinetics / Q000494:pharmacology / Q000633:toxicity; D006626:Hippurates / Q000493:pharmacokinetics / Q000494:pharmacology / Q000633:toxicity; D007200:Indican / Q000493:pharmacokinetics / Q000494:pharmacology / Q000633:toxicity; D007210:Indoleacetic Acids / Q000493:pharmacokinetics / Q000494:pharmacology / Q000633:toxicity; D007668:Kidney / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008657:Metabolic Clearance Rate; D027362:Organic Anion Transport Protein 1 / Q000037:antagonists & inhibitors / Q000378:metabolism*; D027701:Organic Cation Transport Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010400:Penicillin G / Q000493:pharmacokinetics / Q000494:pharmacology*; D011422:Propionates / Q000493:pharmacokinetics / Q000494:pharmacology / Q000633:toxicity; D051381:Rats; D017208:Rats, Wistar; D044967:Serum; D014316:Tritium; D010130:p-Aminohippuric Acid / Q000493:pharmacokinetics / Q000494:pharmacology*; D000099254:Organic Cation Transporter 3
D002250:Carbon Radioisotopes; D004970:Estrone; D005663:Furans; D006626:Hippurates; D007200:Indican; D007210:Indoleacetic Acids; D027362:Organic Anion Transport Protein 1; D027701:Organic Cation Transport Proteins; D010400:Penicillin G; D011422:Propionates; D014316:Tritium; D010130:p-Aminohippuric Acid; D000099254:Organic Cation Transporter 3; C468134:3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid; C421043:Slc22a6 protein, rat; C030737:indoleacetic acid; C017296:estrone sulfate; C030514:hippuric acid