Pharmaceutical Research

, Volume 19, Issue 2, pp 154–161 | Cite as

Proton Gradient-Dependent Transport of Valproic Acid in Human Placental Brush-Border Membrane Vesicles

  • Hiroaki Nakamura
  • Fumihiko Ushigome
  • Noriko Koyabu
  • Shoji Satoh
  • Kiyomi Tsukimori
  • Hitoo Nakano
  • Hisakazu Ohtani
  • Yasufumi SawadaEmail author


Purpose. To investigate the transport mechanism of valproic acid across the human placenta, we used human placental brush-border membrane vesicles and compared them with that of lactic acid.

Methods. Transport of [3H]valproic acid and [14C]lactic acid was measured by using human placental brush-border membrane vesicles.

Results. The uptakes of [3H]valproic acid and [14C]lactic acid into brush-border membrane vesicles were greatly stimulated at acidic extravesicular pH. The uptakes of [3H]valproic acid and [14C]lactic acid were inhibited by various fatty acids, p-chloromercuribenzene sulfonate, α-cyano-4-hydroxycinnamate, and FCCP. A kinetic analysis showed that it was saturable, with Michaelis constants (Kt) of 1.04 ± 0.41 mM and 1.71 ± 0.33 mM for [3H]valproic acid and [14C]lactic acid, respectively. Furthermore, lactic acid competitively inhibited [3H]valproic acid uptake and vice versa.

Conclusion. These results suggest that the transport of valproic acid across the microvillous membrane of human placenta is mediated by a proton-linked transport system that also transports lactic acid. However, some inhibitors differentially inhibited the uptakes of [3H]valproic acid and [14C]lactic acid, suggesting that other transport systems may also contribute to the elevated fetal blood concentration of valproic acid in gravida.

human placenta transport mechanism valproic acid lactic acid 


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  1. 1.
    J. Stulc. Placental transfer of inorganic ions and water. Physiol. Rev. 77:805–836 (1997).Google Scholar
  2. 2.
    W. W. Hay Jr. Placental transport of nutrients to the fetus. Hormone Res. 42:215–222 (1994).Google Scholar
  3. 3.
    A. J. Moe. Placental amino acid transport. Am. J. Physiol. 268:C1321–C1331 (1995).Google Scholar
  4. 4.
    N. A. Reid and C. A. R. Boyd. Further evidence for the presence of two facilitative glucose transporter isoforms in the brush border membrane of the syncytiotrophoblast of the human full term placenta. Biochem. Soc. Trans. 22:267S (1994).Google Scholar
  5. 5.
    S. M. Grassl. Thiamine transport in placental brush border membrane vesicles. Biochim. Biophys. Acta 1371:213–222 (1998).Google Scholar
  6. 6.
    P. D. Prasad, H. Wang, R. Kekuda, T. Fujita, Y. Fei, L. D. Devoe, F. H. Leibach, and V. Ganapathy. Cloning and functional expression of a cDNA encoding a mammalian sodium-dependent vitamin transporter mediating the uptake of pantothenate, biotin, and lipoate. J. Biol. Chem. 273:7501–7506 (1998).Google Scholar
  7. 7.
    J. J. G. Marine, P. Bravo, M. Y. A. El-Mir, and M. A. Serrano. ATP-dependent bile acid transport across microvillous membrane of human term trophoblast. Am. J. Physiol. 268:G685–G694 (1995).Google Scholar
  8. 8.
    F. Ushigome, H. Takanaga, H. Matsuo, S. Yanai, K. Tsukimori, H. Nakano, T. Uchiumi, T. Nakamura, M. Kuwano, H. Ohtani, and Y. Sawada. Human placental transport of vinblastine, vincristine, digoxin and progesterone: contribution of Pglycoprotein. Eur. J. Pharm. 408:1–10 (2000).Google Scholar
  9. 9.
    G. M. Pacifici and R. Nottoli. Placental transfer of drugs administered to the mother. Clin. Pharmacokinetic Concept 28:235–269 (1995).Google Scholar
  10. 10.
    E. M. van der Aa, J. H. J. C. Peereboom-Stegeman, and G. M. Russel. Isolation of syncytial microvillous membrane vesicles from human term placenta and their application in drug-nutrient interaction studies. J. Pharmacol. Toxicol. Metods 34:47–56 (1995).Google Scholar
  11. 11.
    T. Ishizaki, K. Yokochi, K. Chiba, T. Tabuchi, and T. Wagatsuma. Placental transfer of anticonvulsants (phenobarbital, phenytoin, valproic acid) and the elimination from neonates. Pediatric Pharmacol. 1:291–303 (1981).Google Scholar
  12. 12.
    F. D. Malone and M. E. D'Alton. Drugs in pregnancy: anticonvulsants. Semin. Perinatol. 21:114–123 (1997).Google Scholar
  13. 13.
    M. S. Yerby. Teratogenic effects of antiepileptic drugs: what do we advise patients? Epilepsia 38:957–958 (1997).Google Scholar
  14. 14.
    S. R. Alonso de la Torre, M. A. Serrano, F. Alvarado, and J. M. Medina. Carrier-mediated L-lactate transport in brush-border membrane vesicles from rat placenta during late gestation. Biochem. J. 278:535–541 (1991).Google Scholar
  15. 15.
    D. F. Balkovetz, F. H. Leibach, V. B. Mahesh, and V. Ganapathy. A proton gradient is the driving force for uphill transport of lactate in human placental brush-border membrane vesicles. J. Biol. Chem. 263:13823–13830 (1988).Google Scholar
  16. 16.
    N. T. Price, V. N. Jackson, and A. P. Halestrap. Cloning and sequencing of four new mammalian monocarboxylate transporter (MCT) homologues confirms the existence of a transporter family with an ancient past. Biochem. J. 329:321–328 (1998).Google Scholar
  17. 17.
    S. Broer, H. P. Schneider, A. Broer, B. Rahman, B. Hamprecht, and J. W. Deitmer. Characterization of the monocarboxylate transporter 1 expressed in Xenopus laevis oocytes by changes in cytosolic pH. Biochem. J. 333:167–174 (1998).Google Scholar
  18. 18.
    I. Tamai, H. Takanaga, H. Maeda, Y. Sai, T. Ogihara, H. Higashida, and A. Tsuji. Participation of a proton-cotransporter, MCT1 in the intestinal transport of monocarboxylic acids. Biochem. Biophys. Res. Commun. 214:482–489 (1995).Google Scholar
  19. 19.
    C. H. Smith, D. M. Nelson, D. F. King, T. M. Donohue, S. M. Ruzycki, and L. K. Kelley. Characterization of a microvillous membrane preparation from human placental syncytiotrophoblast: a morphologic, biochemical and physiologic study. Am. J. Obstet. Gynecol. 128:190–195 (1977).Google Scholar
  20. 20.
    C. E. Hulstaert, J. L. Torringa, J. Koudstaal, M. J. Hardonk, and I. Molennaar. The characteristic distribution of alkaline phosphatase in full-term human placenta. Gynecol. Invest. 4:23–30 (1973).Google Scholar
  21. 21.
    N. Sawabu, M. Nakagen, T. Wakabayashi, K. Ozaki, D. Toya, N. Hattori, and M. Ishii. gamma-glutamyltranspeptidase as a tumor marker. Nippon Rinsho. 38:4606–4613 (1980).Google Scholar
  22. 22.
    O. H. Lowry, N. J. Rosebrough, A. L. Farr, and R. J. Randall. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193:265–275 (1951).Google Scholar
  23. 23.
    H. Iioka, I. Moriyama, M. Saito, K. Hino, Y. Okamura, and M. Ichijo. Study on changes in placental L-alanine transport activity during gestation. Nippon Sanka Fujinka Gakkai Zasshi 38:529–534 (1986).Google Scholar
  24. 24.
    F. G. M. Russel, P. E. M. van der Linden, W. G. Vermeulen, M. Heijn, C. H. van Os, and C. A. M. van Ginneken. Na+ and H+ gradient-dependent transport of p-aminohippurate in membrane vesicles from dog kidney cortex. Biochem. Phrmacol. 37:2639–2649 (1988).Google Scholar
  25. 25.
    B. Deuticke, E. Beyer, and B. Forst. Discrimination of three parallel pathways of lactate transport in the human erythrocyte membrane by inhibitors and kinetic properties. Biochim. Biophys. Acta 684:96–110 (1982).Google Scholar
  26. 26.
    B. Deuticke, E. Rickert, and E. Beyer. Stereoselective, pHdependent transfer of lactate in mammalian erythrocytes. Biochim. Biophys. Acta 507:137–155 (1978).Google Scholar
  27. 27.
    A. P. Halestrap and R. M. Denton. Specific inhibition of pyruvate transport in rat liver mitochondria and human erythrocytes by a-cyano-4-hydroxycinnamate. Biochem. J. 138:313–316 (1974).Google Scholar
  28. 28.
    R. C. Poole, A. P. Halestrap, S. J. Price, and A. J. Levi. The kinetics of transport of lactate and pyruvate into isolated cardiac myocytes from guinea pig. Biochem. J. 264:409–418 (1989).Google Scholar
  29. 29.
    R. C. Poole, S. L. Cranmer, A. P. Halestrap, and A. J. Levi. Substrate and inhibitor specificity of monocarboxylate transport into heart cells and erythrocytes. Biochem. J. 269:827–829 (1990).Google Scholar
  30. 30.
    R. C. Poole, S. L. Cranmer, D. W. Holdup, and A. P. Halestrap. Inhibition of L-lactate transport and band 3-mediated anion transport in erythrocytes by the novel stilbenedisulphonate N,N,N',N'-tetrabenzyl-4,4-diaminostilbene-2,2'-disulphonate (TBenzDS). Biochim. Biophys. Acta 1070:69–76 (1991).Google Scholar
  31. 31.
    R. C. Poole and A. P. Halestrap. Reversible and irreversible inhibition, by stilbenedisulphonates, of lactate transport into rat erythrocytes. Biochem. J. 275:307–312 (1991).Google Scholar
  32. 32.
    K. Yamaoka, Y. Tanigawara, T. Nakagawa, and T. Uno. A pharmacokinetic analysis program (MULTI) for microcomputer. J. Pharm-Dyn. 4:879–885 (1981).Google Scholar
  33. 33.
    N. L. C. L. Silva, H. Wang, C. V. Harris, D. Singh, and L. Fliegel. Characterization of the Na+/H+ exchanger in human choriocarcinoma (BeWo) cells. Pflugers Archiv Eur. J. Physiol. 433:792–802 (1997).Google Scholar
  34. 34.
    K. Nabuchi, I. Moriyama, M. Akasaki, Y. Katakami, H. Hisanaga, Y. Kato, and M. Ichijo. The study on the human placental L-lactate transport mechanism (using placental microvillous membrane vesicles). Nippon Sanka Fujinka Gakkai Zasshi 41:137–142 (1989).Google Scholar
  35. 35.
    K. D. K. Adkison and D. D. Shen. Uptake of valproic acid into rat brain is mediated by a medium-chain fatty acid transporter. J. Pharmacol. Exp. Ther. 276:1189–1200 (1995).Google Scholar
  36. 36.
    G. L. Edlund and A. P. Halestrap. The kinetics of lactate and pyruvate into rat hepatocytes. Biochem. J. 249:117–126 (1988).Google Scholar
  37. 37.
    A. Ritzhaupt, S. Wood, A. Ellis, K. B. Hosie, and S. P. Shirazi-Beechey. Identification and characterization of a monocarboxylate transporter (MCT1) in pig and human colon: its potential to transport L-lactate as well as butyrate. J. Physiol. 513:719–732 (1998).Google Scholar
  38. 38.
    S. Broer, A. Broer, H. P. Schneider, C. Stegen, and A. P. Halestrap. Characterization of the high-affinity monocarboxylate transporter MCT2 in Xenopus laevis oocytes. Biochem. J. 341: 529–535 (1999).Google Scholar

Copyright information

© Plenum Publishing Corporation 2002

Authors and Affiliations

  • Hiroaki Nakamura
    • 1
  • Fumihiko Ushigome
    • 1
  • Noriko Koyabu
    • 1
  • Shoji Satoh
    • 2
  • Kiyomi Tsukimori
    • 1
  • Hitoo Nakano
    • 1
  • Hisakazu Ohtani
    • 1
  • Yasufumi Sawada
    • 1
    Email author
  1. 1.Department of Medico-Pharmaceutical Sciences, Graduate School of Pharmaceutical SciencesKyushu UniversityHigashi-ku, FukuokaJapan
  2. 2.Department of Reproduction and Gynecology, Graduate School of Medical SciencesKyushu UniversityHigashi-ku, FukuokaJapan

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