1. Academic Validation
  2. Nelfinavir suppresses insulin signaling and nitric oxide production by human aortic endothelial cells: protective effects of thiazolidinediones

Nelfinavir suppresses insulin signaling and nitric oxide production by human aortic endothelial cells: protective effects of thiazolidinediones

  • Ochsner J. 2013 Spring;13(1):76-90.
Debasis Mondal 1 Kai Liu Milton Hamblin Joseph A Lasky Krishna C Agrawal
Affiliations

Affiliation

  • 1 Department of Pharmacology and.
PMID: 23533049
Abstract

Background: In human immunodeficiency virus 1 (HIV-1)-infected individuals, exposure to a protease inhibitor (PI)-based highly active antiretroviral therapy (HAART) regimen increases Cardiovascular Disease and endothelial dysfunction. However, the mechanisms of PI-induced effects on endothelial cells (ECs) are not known. Furthermore, strategies to suppress these deleterious outcomes of PIs need to be developed. Insulin-induced PI3K/Akt signaling and endothelial nitric oxide (NO)-synthase (eNOS) phosphorylation regulates NO production by ECs that maintain vascular homeostasis. We evaluated whether nelfinavir (NEL), a potent HIV-1 PI that suppresses Akt phosphorylation, can alter insulin-induced NO production in human aortic endothelial cells (HAECs) and whether Insulin sensitization of HAECs via the peroxisome proliferator-activated receptor-gamma agonists, thiazolidinediones, can ameliorate these side effects.

Methods: Real-time NO production in HAECs was monitored by fluorimetric dyes DAF-FM DA and DAF-2 DA. Immunodetection studies were used to determine the phosphorylation of Akt, eNOS, Insulin receptor-β (IR-β), Insulin Receptor substrate-1 (IRS-1), and PI3K/p85α. Expression of eNOS messenger RNA was measured by reverse transcription polymerase chain reaction.

Results: In vitro exposure (72 hours) of HAECs to NEL (0.25-2 μg/mL) decreased both basal (2.5-fold) and insulin-induced NO production (4- to 5-fold). NEL suppressed insulin-induced phosphorylation of both Akt and eNOS at serine residues 473 and 1177, respectively. NEL decreased tyrosine phosphorylation of IR-β, IRS-1, and PI3K. Coexposure to troglitazone (TRO; 250 nM) ameliorated the suppressive effects of NEL on Insulin signaling and NO production. Coexposure to TRO also increased eNOS expression in NEL-treated HAECs.

Conclusion: Our findings indicate that treatment with potent Insulin sensitizers may protect against PI-mediated endothelial dysfunction during long-term HAART.

Keywords

Endothelial cells; HIV-1; nelfinavir; nitric oxide; thiazolidinediones.

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