Transport of monocarboxylic acids at the blood-brain barrier: studies with monolayers of primary cultured bovine brain capillary endothelial cells.
The kinetics and mechanism of the transport of monocarboxylic acids (MCAs) were studied by using primary cultured bovine brain capillary endothelial cells. Concentration-dependent uptake of acetic acid was observed, and the kinetic parameters were estimated as follows: the Michaelis constant, Kt, was 3.41 +/- 1.87 mM, the maximum uptake rate, Jmax, was 144.7 +/- 55.7 nmol/mg of protein/min and the nonsaturable first-order rate constant, Kd, was 6.66 +/- 1.98 microliters/mg of protein/min. At medium pH below 7.0, the uptake rate of [3H]acetic acid increased markedly with decreasing medium pH, whereas pH-independent uptake was observed in the presence of 10 mM acetic acid. An energy requirement for [3H]acetic acid uptake was also demonstrated, because metabolic inhibitors (2,4-dinitrophenol and rotenone) reduced significantly the uptake rate (P less than .05). Carbonylcyanide-p-trifluoro-methoxyphenylhydrazone, a protonophore, inhibited significantly the uptake of [3H]acetic acid at medium pH of 5.0 and 6.0, whereas 4,4'-diisothiocyanostilben-2,2'-disulfonic acid did not. Several MCAs inhibited significantly the uptake rate of [3H]acetic acid, whereas di- and tricarboxylic acids did not. The uptake of [3H]acetic acid was competitively inhibited by salicylic acid, with an inhibition constant, Ki, of 3.60 mM, suggesting a common transport system between acetic acid and salicylic acid. Moreover, at the medium pH of 7.4, salicylic acid and valproic acid inhibited significantly the uptake of [3H]acetic acid, demonstrating that the transport of MCA drugs could also be ascribed to the MCA transport system at the physiologic pH.(ABSTRACT TRUNCATED AT 250 WORDS)
Terasaki
T
T
Department of Pharmaceutics, Faculty of Pharmaceutical Sciences, Kanazawa University, Japan.
Takakuwa
S
S
Moritani
S
S
Tsuji
A
A
D019325:3-O-Methylglucose; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000031:analogs & derivatives*; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D000085:Acetates / Q000493:pharmacokinetics; D019342:Acetic Acid; D000596:Amino Acids / Q000494:pharmacology; D000818:Animals; D001812:Blood-Brain Barrier / Q000502:physiology*; D001921:Brain / Q000098:blood supply / Q000166:cytology / Q000378:metabolism; D002196:Capillaries / Q000166:cytology / Q000378:metabolism; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000494:pharmacology; D002264:Carboxylic Acids / Q000493:pharmacokinetics*; D002417:Cattle; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007930:Leucine / Q000493:pharmacokinetics; D008757:Methylglucosides / Q000493:pharmacokinetics; D012459:Salicylates / Q000494:pharmacology; D020156:Salicylic Acid; D014316:Tritium; D014635:Valproic Acid / Q000494:pharmacology
D000085:Acetates; D000596:Amino Acids; D002264:Carboxylic Acids; D008757:Methylglucosides; D012459:Salicylates; D014316:Tritium; D019325:3-O-Methylglucose; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; C018564:dihydro-DIDS; D014635:Valproic Acid; D007930:Leucine; D020156:Salicylic Acid; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid; D019342:Acetic Acid