Modulation of sodium-coupled uptake and membrane fluidity by cisplatin in renal proximal tubular cells in primary culture and brush-border membrane vesicles.
The proximal tubule appears to be the main target for the adverse effects of cis-diamminedichloroplatinum (II) (cDDP). We evaluated the early effects of cDDP at concentrations (3 to 67 microM) lower that those which alter cell viability, on three apical transport systems and on the physical state of the brush border membrane (BBM) in rabbit proximal tubule (RPT) cells in primary culture. The maximal effect, corresponding to a 30% decrease in Na(+)-coupled uptake of phosphate (Pi) and alpha-methylglucopyranoside (MGP) and a twofold increase in Na(+)-coupled alanine uptake, was obtained at 17 microM (5 micrograms/ml) cDDP and occurred through a modification of their affinity. At this concentration, cDDP increased BBM fluidity and decreased the BBM cholesterol content by 28%, without increasing the permeability of tight junctions. To clarify the role of cDDP-induced increase in BBM fluidity on alterations of Na(+)-coupled uptake, these parameters were also investigated in BBM vesicles isolated from rabbit renal cortex directly exposed to cDDP. cDDP induced a concentration-dependent inhibition of Na(+)-coupled uptake of MGP, Pi and alanine in BBM vesicles from the renal cortex, associated with a decrease in protein sulfhydryl content, without modifying BBM fluidity. Our findings strongly suggest that the cDDP-induced increase in BBM fluidity in RPT cells results from an indirect mechanism, possibly an alteration of cholesterol metabolism, and did not play a major role in the cDDP-induced inhibition of Na+/Pi and Na+/glucose cotransport systems that may be mainly mediated through a direct chemical interaction with essential sulfhydryl groups of the transporters.
Courjault-Gautier
F
F
Département Sécurité du Médicament, Rhône-Poulenc Rorer SA, Vitry sur Seine, France.
Le Grimellec
C
C
Giocondi
M C
MC
Toutain
H J
HJ
D000409:Alanine / Q000378:metabolism; D000818:Animals; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism; D002463:Cell Membrane Permeability / Q000187:drug effects; D002478:Cells, Cultured / Q000187:drug effects; D002945:Cisplatin / Q000494:pharmacology*; D017136:Ion Transport / Q000187:drug effects; D007668:Kidney / Q000187:drug effects / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D015227:Lipid Peroxidation / Q000187:drug effects; D008560:Membrane Fluidity / Q000187:drug effects*; D008757:Methylglucosides / Q000378:metabolism; D008871:Microvilli / Q000187:drug effects* / Q000378:metabolism; D010710:Phosphates / Q000378:metabolism; D011506:Proteins / Q000187:drug effects; D011817:Rabbits; D012964:Sodium / Q000378:metabolism*; D013438:Sulfhydryl Compounds / Q000378:metabolism
D008757:Methylglucosides; D010710:Phosphates; D011506:Proteins; D013438:Sulfhydryl Compounds; C027020:methylglucoside; D012964:Sodium; D000409:Alanine; D002945:Cisplatin