MRP (ABCC) transporters-mediated efflux of anti-HIV drugs, saquinavir and zidovudine, from human endothelial cells

Exp Biol Med (Maywood). 2008 Sep;233(9):1149-60. doi: 10.3181/0802-RM-59. Epub 2008 Jun 5.


The constituents of highly active anti-retroviral therapy (HAART) include HIV-1 protease inhibitors (HPIs) and nucleoside reverse transcriptase inhibitors (NRTIs). Endothelial cell (EC) barriers, especially the blood-brain-barrier (BBB) suppresses the entry of HAART drugs to subendothelial HIV-1 reservoirs. The ATP binding cassette (ABC) transporter family members, multidrug resistant-1 (MDR-1) and multidrug resistance-associated proteins (MRPs) can efflux both HPIs and NRTIs from intracellular compartments. Using brain derived ECs from non-human sources, previous studies suggested a dominant role for MDR-1 in HAART efflux from the BBB. However, due to species variations in ABC-transporter expression, drug-efflux functions using human brain ECs need to be investigated. Furthermore, roles of ABC-transporters in drug-efflux from systemic EC barriers need to be studied. We monitored the expression of ABC-transporters in primary human ECs obtained from brain (HBMVECs), aorta (HAECs), pulmonary-artery (HPAECs), dermal-microvessel (HDMVECs) and umbilical vein (HUVECs). Gene expression for MDR-1 and MRPs (MRP-1 to MRP-5) were analyzed by reverse transcriptase polymerase chain reaction (RT-PCR). Drug efflux functions were determined by calcein retention assays. Intracellular accumulation of both 3H-saquinavir (an HPI) and 3H-zidovudine (an NRTI) were also monitored in HAECs and HBMVECs. Both assays were carried out in presence of verapamil (20-60 microM) or MK-571 (12.5-50 microM) inhibitors of MDR-1 and MRPs, respectively in presence of verapamil or MK-571. The HBMVECs expressed higher levels of MRPs than MDR-1 and only MK-571 significantly (P<0.01) suppressed calcein efflux from these cells. However, both HAECs and HPAECs showed MDR-1 and MRP expression and calcein efflux was inhibited by both verapamil and MK-571. Both inhibitors suppressed 3H-saqubinavir efflux from HAECs, but only MK-571 suppressed saquinavir efflux from HBMVECs. In both ECs, 3H-zidovudine efflux was only suppressed by MK-571. Thus, primary human ECs, especially brain derived ECs, predominantly express MRPs and their specific inhibition may enhance HAART efficacy in subendothelial HIV-1 reservoirs.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Anti-HIV Agents / metabolism*
  • Aorta / metabolism
  • Brain / metabolism
  • Cells, Cultured
  • Endothelial Cells / metabolism*
  • Fluoresceins / metabolism
  • Gene Expression Regulation
  • Humans
  • Multidrug Resistance-Associated Proteins / genetics
  • Multidrug Resistance-Associated Proteins / metabolism*
  • Saquinavir / metabolism*
  • Zidovudine / metabolism*


  • Anti-HIV Agents
  • Fluoresceins
  • Multidrug Resistance-Associated Proteins
  • Zidovudine
  • Saquinavir
  • fluorexon