Primary culture of rabbit proximal tubules as a cellular model to study nephrotoxicity of xenobiotics.
The effects of gentamicin treatment on functions of the plasma membrane-bound proteins in situ were investigated in primary culture of rabbit proximal tubular cells (PTC), a recognized model of renal epithelial cells. Activities of apical and basolateral enzymes, activities of phosphate, glucose and alanine sodium-coupled transport systems and leakage of the cytosolic enzyme lactate dehydrogenase (LDH) were determined in PTC grown in glucose-free culture medium as confluent monolayers and incubated with the aminoglycoside. Gentamicin altered in a concentration- and time-dependent manner the activity of dipeptidyl peptidase IV (DPP IV), neutral aminopeptidase (NAP), Na+K(+)-ATPase and the Vmax of sodium-dependent glucose and phosphate uptake, whereas gamma-glutamyl-transpeptidase (GGT) and sodium-dependent alanine uptake were unaffected. Identical concentration of gentamicin was required to induce LDH leakage and cell functions impairment. In contrast, under short time exposure, a condition where the enzyme activities were untouched, mercuric chloride inhibited to a similar extent the activity of the three sodium-coupled transport systems. These data suggest that whereas alterations in membrane fluidity might mediate the effects of gentamicin on membrane functions, the inhibition of transports by mercuric chloride rather reflects an effect on sodium permeability of the apical membrane. They also suggest that study of Na(+)-coupled transports in proximal tubular cells grown in primary culture is a simple and sensitive in vitro model to assess drug-induced nephrotoxicity.
Blais
A
A
Institut National de la Santé et de la Recherche Médicale U 251, Département de Physiologie, Faculté de Médecine Xavier Bichat, Paris, France.
Morvan-Baleynaud
J
J
Friedlander
G
G
Le Grimellec
C
C
D000626:Aminopeptidases / Q000378:metabolism; D000818:Animals; D001693:Biological Transport, Active / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D018819:Dipeptidyl Peptidase 4; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000378:metabolism; D005839:Gentamicins / Q000494:pharmacology; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D007700:Kinetics; D008757:Methylglucosides / Q000493:pharmacokinetics; D008954:Models, Biological; D011817:Rabbits; D012964:Sodium / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D015262:Xenobiotics / Q000633:toxicity*
D005839:Gentamicins; D008757:Methylglucosides; D015262:Xenobiotics; C027020:methylglucoside; D012964:Sodium; D000626:Aminopeptidases; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases; D018819:Dipeptidyl Peptidase 4; D000254:Sodium-Potassium-Exchanging ATPase