title: Highly active antiretroviral therapy drug combination induces oxidative stress and mitochondrial dysfunction in immortalized human blood-brain barrier endothelial cells.
abstract: The era of highly active antiretroviral therapy (HAART) has controlled AIDS and its related disorders considerably; however, the prevalence of HIV-1-associated neurocognitive disorders has been on the rise in the post-HAART era. In view of these developments, we investigated whether a HAART drug combination of 3'-azido-2',3'-deoxythymidine (AZT) and indinavir (IDV) can alter the functionality of the blood-brain barrier (BBB) endothelial cells, thereby exacerbating this condition. The viability of hCMEC/D3 cells (in vitro model of BBB) that were exposed to these drugs was significantly reduced after 72h treatment, in a dose-dependent manner. Reactive oxygen species were highly elevated after the exposure, indicating that mechanisms that induce oxidative stress were involved. Measures of oxidative stress parameters, such as glutathione and malondialdehyde, were altered in the treated groups. Loss of mitochondrial membrane potential, as assessed by fluorescence microscopy and decreased levels of ATP, indicated that cytotoxicity was mediated through mitochondrial dysfunction. Furthermore, AZT+IDV treatment caused apoptosis in endothelial cells, as assessed by the expression of cytochrome c and procaspase-3 proteins. Pretreatment with the thiol antioxidant N-acetylcysteine amide reversed some of the pro-oxidant effects of AZT+IDV. Results from our in vitro studies indicate that the AZT+IDV combination may affect the BBB in HIV-infected individuals treated with HAART drugs.
authors.affiliation: Department of Chemistry, Missouri University of Science and Technology, Rolla, MO 65409, USA.
authors.forename: Kalyan Reddy
authors.initials: KR
authors.lastname: Manda
authors.forename: Atrayee
authors.initials: A
authors.lastname: Banerjee
authors.forename: William A
authors.initials: WA
authors.lastname: Banks
authors.forename: Nuran
authors.initials: N
authors.lastname: Ercal
mesh_terms: D000111:Acetylcysteine / Q000031:analogs & derivatives / Q000494:pharmacology / Q000627:therapeutic use; D000255:Adenosine Triphosphate / Q000032:analysis; D023241:Antiretroviral Therapy, Highly Active / Q000009:adverse effects*; D017209:Apoptosis / Q000187:drug effects; D001812:Blood-Brain Barrier / Q000378:metabolism; D053148:Caspase 3 / Q000032:analysis / Q000096:biosynthesis; D002461:Cell Line, Transformed; D045304:Cytochromes c / Q000032:analysis / Q000096:biosynthesis; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D005978:Glutathione / Q000032:analysis; D015658:HIV Infections / Q000188:drug therapy / Q000378:metabolism; D017320:HIV Protease Inhibitors / Q000009:adverse effects / Q000494:pharmacology; D015497:HIV-1 / Q000378:metabolism; D006801:Humans; D019469:Indinavir / Q000009:adverse effects / Q000494:pharmacology; D008315:Malondialdehyde / Q000032:analysis; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D017382:Reactive Oxygen Species / Q000378:metabolism; D015215:Zidovudine / Q000009:adverse effects / Q000494:pharmacology
chemical_list: D017320:HIV Protease Inhibitors; D017382:Reactive Oxygen Species; D015215:Zidovudine; C487056:N-Acetylcysteinamide; D008315:Malondialdehyde; D019469:Indinavir; D000255:Adenosine Triphosphate; D045304:Cytochromes c; D053148:Caspase 3; D005978:Glutathione; D000111:Acetylcysteine