The characterisation and uptake of paraquat in cultured baboon kidney proximal tubule cells (bPTC).
A primary culture of baboon proximal tubule cells (bPTC) was prepared and characterised using LLC-PK1 cells of proximal tubule origin and MDCK cells of distal tubule origin, as positive and negative references, respectively. The proximal tubular origin of the bPTC was determined by morphological studies, immunoperoxidase staining and the expression of proximal tubule markers alkaline phosphatase and gammaglutamyltransferase. The hypothesis that paraquat (PQ) is transported by the bPTC was investigated. The cytotoxic threshold for PQ in these cells was determined and compared to the LLC-PK1 and MDCK cells. Furthermore, this study investigated the transport of the monovalent cation tetraethyl ammonium (TEA) and the polyvalent cation cimetidine in the bPTC and demonstrated their effect on the cellular uptake of PQ. The cytotoxic threshold of PQ in the bPTC, determined by cellular viability studies using the method of Trypan blue exclusion, is 0.05 mM at 2 h incubation. The LC50 after 24 h is 76, 61 and 455 microM for the bPTC, LLC-PK1 and MDCK cells, respectively. This indicates that proximal tubule cells are more susceptible to PQ toxicity compared to distal tubule cells, which is consistent with clinical PQ toxicity where renal damage is found predominantly in the proximal renal tubules. The cations PQ and cimetidine were actively transported by the bPTC. The uptake of PQ (0.05 mM) commenced after 15 min whereas cimetidine (0.5 mM) uptake was evident after 2 min. Furthermore, cimetidine was shown to compete with PQ for uptake in the bPTC. Coincubating PQ (0.05 mM) and cimetidine (0.5 mM) for 60 min resulted in an approximate 50% decrease in PQ uptake. The cation TEA was not transported by the bPTC suggesting either a genetic mutation or complete absence of the transporter for TEA in the cells. The results suggest that PQ may be transported by the same cation transporter as cimetidine and not TEA, indicating PQ uptake in the bPTC to be via a polyvalent organic cation transporter.
Machaalani
R
R
Department of Renal Medicine and Toxicology Unit, Royal Prince Alfred Hospital, Camperdown, Australia.
Lazzaro
V
V
Duggin
G G
GG
D000276:Adjuvants, Immunologic / Q000493:pharmacokinetics; D000818:Animals; D001692:Biological Transport / Q000187:drug effects; D002412:Cations / Q000493:pharmacokinetics; D002460:Cell Line; D002478:Cells, Cultured; D002927:Cimetidine / Q000493:pharmacokinetics; D004285:Dogs; D006540:Herbicides / Q000493:pharmacokinetics* / Q000633:toxicity; D007686:Kidney Tubules, Distal / Q000187:drug effects / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010215:Papio; D010269:Paraquat / Q000493:pharmacokinetics* / Q000633:toxicity; D013552:Swine; D019789:Tetraethylammonium / Q000493:pharmacokinetics
D000276:Adjuvants, Immunologic; D002412:Cations; D006540:Herbicides; D019789:Tetraethylammonium; D002927:Cimetidine; D010269:Paraquat