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Mitochondrial Mechanisms of Nelfinavir Toxicity in Human Brain Microvascular Endothelial cells
Author(s) -
Unis Graham,
Baker Tyler P,
Rajaprabhakaran Gowthamram,
Sure Venkata N,
Jain Neelesh P,
Abraham Vihas M,
Peterson Nicholas R,
Gordon Angellica O,
Chen Allen L,
Cama Zeinab,
Mondal Debasis,
Katakam Prasad VG
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.953.4
Subject(s) - nelfinavir , viability assay , oxidative stress , pharmacology , reactive oxygen species , chemistry , mitochondrion , superoxide , medicine , biochemistry , apoptosis , immunology , viral load , human immunodeficiency virus (hiv) , antiretroviral therapy , enzyme
Antiretroviral therapies with combinations of mechanistically different classes of drugs resulted in reduced HIV‐associated neurocognitive disorders, however, milder forms of cognitive disorders continue to be disabling in AIDS patients. Toxicity resulting from long‐term use of antiretroviral drugs have been implicated in the cognitive disorders but the exact underlying mechanisms are poorly understood. We hypothesized that nelfinavir, a protease inhibitor, causes the disruption of blood brain barrier (BBB) by promoting mitochondrial oxidative stress and impairing mitochondrial respiration. Our studies examined the effects of nelfinavir in human brain microvascular endothelial cells (BMECs). Cell viability following 48 h Nelfinavir treatment of hBMECs was determined using CCK‐8 assay. Measurements of superoxide levels were made by electron spin resonance spectrometery using spin probes for total (1‐Hydroxy‐3‐methoxycarbonyl‐2,2,5,5‐tetramethyl‐pyrrolidine) and mitochondrial superoxide (mito‐Tempo‐H). Oxygen consumption rates (OCR) were measured by Seahorse Analyzer. Transcellular endocytosis measurements were performed by using 40 kDa‐FITC‐dextran transfer in BMECs grown in transwells. Nelfinavir treatment caused an increase in cell proliferation of BMECs at low doses (0.5 μM‐1.5 μM) but decreased viability at high doses (5 μM‐10 μM). Nelfinavir at therapeutic doses promoted the generation of total and mitochondrial superoxide in BMECs. In addition, nelfinavir dose‐dependently diminished the basal OCR and reserve mitochondrial respiratory capacity in BMECs. Moreover, nelfinavir dose‐dependently promoted increased transcellular permeability in BMECs. Thus, nelfinavir induced mitochondrial oxidative stress promotes diminished mitochondrial respiratory capacity and decreased cell proliferation leading to the disruption of BBB. Support or Funding Information Support: PVK: Louisiana BoRSF‐RCS (LEQSF(2014‐17)‐RD‐A‐11) and AHA Scientist Development Grant (14SDG20490359); IR: AHA Postdoctoral Fellowship Grant (15POST23040005); and DWB: NIH grants (HL‐077731and HL093554).