NAC Attenuates LPS-Induced Toxicity in Aspirin-Sensitized Mouse Macrophages via Suppression of Oxidative Stress and Mitochondrial Dysfunction

Abstract

Bacterial endotoxin lipopolysaccharide (LPS) induces the production of inflammatory cytokines and reactive oxygen species (ROS) under in vivo and in vitro conditions. Acetylsalicylic acid (ASA, aspirin) is a commonly used anti-inflammatory drug. Our aim was to study the effects of N-acetyl cysteine (NAC), an antioxidant precursor of GSH synthesis, on aspirin-sensitized macrophages treated with LPS. We investigated the effects of LPS alone and in conjunction with a sub-toxic concentration of ASA, on metabolic and oxidative stress, apoptosis, and mitochondrial function using J774.2 mouse macrophage cell line. Protection from LPS-induced toxicity by NAC was also studied. LPS alone markedly induced ROS production and oxidative stress in macrophage cells. When ASA was added to LPS-treated macrophages, the increase in oxidative stress was significantly higher than that with LPS alone. Similarly, alteration in glutathione-dependent redox metabolism was also observed in macrophages after treatment with LPS and ASA. The combination of LPS and ASA selectively altered the CYP 3A4, CYP 2E1 and CYP 1A1 catalytic activities. Mitochondrial respiratory complexes and ATP production were also inhibited by LPS-ASA treatment. Furthermore a higher apoptotic cell death was also observed in LPS-ASA treated macrophages. NAC pre-treatment showed protection against oxidative stress induced apoptosis and mitochondrial dysfunction. These effects are presumed, at least in part, to be associated with alterations in NF-κB/Nrf-2 mediated cell signaling. These results suggest that macrophages are more sensitive to LPS when challenged with ASA and that NAC pre-treatment protects the macrophages from these deleterious effects.

Oxidative status in treatment-naïve essential thrombocythemia: a pilot study in a single center

“…Finally, considering antiplatelet treatment, previous in vitro model studies demonstrated the implications of altered glutathione metabolism, ROS levels, and DNA damages in aspirin‐induced OS . In contrast with Raza et al , we did not find any difference in ROS, GSH, GSSG, and 8‐OHdG levels between patients treated with antiplatelet drugs and not treated patients.…”

Histomorphological evaluations on the frontal cortex extrapyramidal cell layer following administration of N-Acetyl cysteine in aluminum induced neurodegeneration rat model

“…Aluminum caused extensive expression of astrocytes (gliosis) a condition seen in neurodegenerative conditions such as AD and PD (Raza et al 2014;Sharhripour et al, 2014) in Aluminum treated (Fig.5B) This is attributed to a sustained inflammatory response a crucial fact for the progression of neuron loss in AD (Wang et al 2015), this is term neuro-inflammation characterized by specialized response by activated glial cells (reactive gliosis) an oxidative stress mechanism (McGeer and McGeer 2008;Miller et al 2009). According to Deepmala et al (2015) and Batlle et al (2015) astrocytes are active during neuroinflammation to help neuron repair, support it during injury or toxic assault and restore cerebral homeostasis.…”

Cytoprotective Effects of N-Acetylcysteine on Streptozotocin- Induced Oxidative Stress and Apoptosis in RIN-5F Pancreatic β-Cells

“…For this, cells were incubated with 5μM DCFDA for 30min at 37°C. Cells were washed twice with PBS, trypsinized, resuspended in 3ml PBS and fluorescence immediately analyzed by flow cytometry as described before [31].…”

“…The total and oxidized glutathione are then measured from the standard curve and the GSH/GSSG ratios calculated. GSH-Px activity using cumene hydroperoxide [34] and GSH-reductase using oxidized glutathione (GSSG) [35] as substrates were measured by standard protocols as described before [31,33,36,37].…”