Cationic uremic toxins affect human renal proximal tubule cell functioning through interaction with the organic cation transporter
(2013) In Pflügers Archiv 465(12). p.1701-1714- Abstract
- Several organic cations, such as guanidino compounds and polyamines, have been found to accumulate in plasma of patients with kidney failure due to inadequate renal clearance. Here, we studied the interaction of cationic uremic toxins with renal organic cation transport in a conditionally immortalized human proximal tubule epithelial cell line (ciPTEC). Transporter activity was measured and validated in cell suspensions by studying uptake of the fluorescent substrate 4-(4-(dimethylamino)styryl)-N-methylpyridinium-iodide (ASP(+)). Subsequently, the inhibitory potencies of the cationic uremic toxins, cadaverine, putrescine, spermine and spermidine (polyamines), acrolein (polyamine breakdown product), guanidine, and methylguanidine (guanidino... (More)
- Several organic cations, such as guanidino compounds and polyamines, have been found to accumulate in plasma of patients with kidney failure due to inadequate renal clearance. Here, we studied the interaction of cationic uremic toxins with renal organic cation transport in a conditionally immortalized human proximal tubule epithelial cell line (ciPTEC). Transporter activity was measured and validated in cell suspensions by studying uptake of the fluorescent substrate 4-(4-(dimethylamino)styryl)-N-methylpyridinium-iodide (ASP(+)). Subsequently, the inhibitory potencies of the cationic uremic toxins, cadaverine, putrescine, spermine and spermidine (polyamines), acrolein (polyamine breakdown product), guanidine, and methylguanidine (guanidino compounds) were determined. Concentration-dependent inhibition of ASP(+) uptake by TPA, cimetidine, quinidine, and metformin confirmed functional endogenous organic cation transporter 2 (OCT2) expression in ciPTEC. All uremic toxins tested inhibited ASP(+) uptake, of which acrolein required the lowest concentration to provoke a half-maximal inhibition (IC50 = 44 +/- 2 mu M). A Dixon plot was constructed for acrolein using three independent inhibition curves with 10, 20, or 30 mu M ASP(+), which demonstrated competitive or mixed type of interaction (K (i) = 93 +/- 16 mu M). Exposing the cells to a mixture of cationic uremic toxins resulted in a more potent and biphasic inhibitory response curve, indicating complex interactions between the toxins and ASP(+) uptake. In conclusion, ciPTEC proves a suitable model to study cationic xenobiotic interactions. Inhibition of cellular uptake transport was demonstrated for several uremic toxins, which might indicate a possible role in kidney disease progression during uremia. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/4273397
- author
- Schophuizen, Carolien M. S. ; Wilmer, Martijn J. ; Jansen, Jitske ; Gustavsson, Lena LU ; Hilgendorf, Constanze ; Hoenderop, Joost G. J. ; van den Heuvel, Lambert P. and Masereeuw, Rosalinde
- organization
- publishing date
- 2013
- type
- Contribution to journal
- publication status
- published
- subject
- keywords
- Human proximal tubule cell, OCT, Uremic toxin, Polyamines, Guanidine, Acrolein
- in
- Pflügers Archiv
- volume
- 465
- issue
- 12
- pages
- 1701 - 1714
- publisher
- Springer
- external identifiers
-
- wos:000327409000004
- scopus:84890433586
- pmid:23812163
- ISSN
- 0031-6768
- DOI
- 10.1007/s00424-013-1307-z
- language
- English
- LU publication?
- yes
- additional info
- The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Molecular Medicine (013031200)
- id
- 784fc9da-ac06-42fa-ba5c-1e9377947939 (old id 4273397)
- date added to LUP
- 2016-04-01 14:38:17
- date last changed
- 2022-04-22 04:24:14
@article{784fc9da-ac06-42fa-ba5c-1e9377947939,
abstract = {{Several organic cations, such as guanidino compounds and polyamines, have been found to accumulate in plasma of patients with kidney failure due to inadequate renal clearance. Here, we studied the interaction of cationic uremic toxins with renal organic cation transport in a conditionally immortalized human proximal tubule epithelial cell line (ciPTEC). Transporter activity was measured and validated in cell suspensions by studying uptake of the fluorescent substrate 4-(4-(dimethylamino)styryl)-N-methylpyridinium-iodide (ASP(+)). Subsequently, the inhibitory potencies of the cationic uremic toxins, cadaverine, putrescine, spermine and spermidine (polyamines), acrolein (polyamine breakdown product), guanidine, and methylguanidine (guanidino compounds) were determined. Concentration-dependent inhibition of ASP(+) uptake by TPA, cimetidine, quinidine, and metformin confirmed functional endogenous organic cation transporter 2 (OCT2) expression in ciPTEC. All uremic toxins tested inhibited ASP(+) uptake, of which acrolein required the lowest concentration to provoke a half-maximal inhibition (IC50 = 44 +/- 2 mu M). A Dixon plot was constructed for acrolein using three independent inhibition curves with 10, 20, or 30 mu M ASP(+), which demonstrated competitive or mixed type of interaction (K (i) = 93 +/- 16 mu M). Exposing the cells to a mixture of cationic uremic toxins resulted in a more potent and biphasic inhibitory response curve, indicating complex interactions between the toxins and ASP(+) uptake. In conclusion, ciPTEC proves a suitable model to study cationic xenobiotic interactions. Inhibition of cellular uptake transport was demonstrated for several uremic toxins, which might indicate a possible role in kidney disease progression during uremia.}},
author = {{Schophuizen, Carolien M. S. and Wilmer, Martijn J. and Jansen, Jitske and Gustavsson, Lena and Hilgendorf, Constanze and Hoenderop, Joost G. J. and van den Heuvel, Lambert P. and Masereeuw, Rosalinde}},
issn = {{0031-6768}},
keywords = {{Human proximal tubule cell; OCT; Uremic toxin; Polyamines; Guanidine; Acrolein}},
language = {{eng}},
number = {{12}},
pages = {{1701--1714}},
publisher = {{Springer}},
series = {{Pflügers Archiv}},
title = {{Cationic uremic toxins affect human renal proximal tubule cell functioning through interaction with the organic cation transporter}},
url = {{http://dx.doi.org/10.1007/s00424-013-1307-z}},
doi = {{10.1007/s00424-013-1307-z}},
volume = {{465}},
year = {{2013}},
}