Abstract
OBJECTIVE: HIV protease inhibitors have been successfully used in highly active antiretroviral therapy of HIV-1 infection, but their benefits are compromised by a number of clinically important adverse side-effects. Several studies showed that protease inhibitors induce sarco/endoplasmic reticulum stress and overproduction of reactive oxygen species (ROS), but the hierarchy of these events was never established in protease inhibitor-treated cells. Our objective was to determine whether ROS production and sarco/endoplasmic reticulum stress were co-induced by protease inhibitors in human primary skeletal myotubes and whether antioxidant treatment with resveratrol could protect against protease inhibitor-induced cellular damages. DESIGN AND METHODS: We analyzed in cultures of primary human skeletal myotubes the effects of four protease inhibitors (atazanavir, lopinavir, ritonavir and saquinavir) on ROS production (by measuring the reduction of nitro blue tetrazolium and by fluorescence microscopy using CM-H2DCFDA), on sarco/endoplasmic reticulum stress induction (by immunofluorescence and electron microscopy) and on the expression and localization at lipid rafts of Caveolin 3 and Flotillin 1, two major components of lipid rafts (by immunoblotting and biochemical experiments). Cells were co-incubated with resveratrol to assess its protective effects. RESULTS: In myotubes, protease inhibitors increased ROS production, altered sarco/endoplasmic reticulum morphology, increased expression of C/EBP homologous protein, a sarco/endoplasmic reticulum stress marker, and decreased expression and localization at lipid rafts of Caveolin 3 and Flotillin 1. Importantly, these deleterious protease inhibitor effects were inhibited by the antioxidant resveratrol. CONCLUSION: Our findings demonstrate a protective effect of resveratrol against protease inhibitor-induced sarco/endoplasmic reticulum stress in human myotubes.