Investigation of Acetaminophen Induction Effects on GSH Concentration by N-Acetylcysteine and 2-Oxothrazolidin-4-Carboxylate (OTC) and Its Protective Effect on sulfure Mustard Injures in Hf2ff Cells
Trauma Monthly: 10 (3); 175-182 Article Type: Research Article
L. Investigation of Acetaminophen Induction Effects on GSH Concentration by N-Acetylcysteine and 2-Oxothrazolidin-4-Carboxylate (OTC) and Its Protective Effect on sulfure Mustard Injures in Hf2ff Cells,
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Objective: The NAC and OTC effects in the presence of acetaminophen as an oxidative stress on GSH concentration in cultured HF2FF cells were studied . The protective effect of GSH on sulfur mustard injuries in cultured cells was also investigated. Materials and Methods: In this research the effects of N-acetlcysteine (NAC) and OTC with presence of an inducer such as acetaminophen on intracellular Glutathione (GSH) and g -GCS and GSSGR activities in induced hemogenate were investigated. After induction of GSH concentration in cells, induced, normal and control cells were expressed with 180 m M HD for 1h and after washing, fresh media were added and incubated 24h, the cell viability was investigated by the GV method. Results: The addition of 0.1mM NAC to HF2FF cells incubated with 25mM acetaminophen increased the GSH, g -GCS and GSSGR activities respectively from 0.331±0.12 μM/mg protein to 0.784±0.11 μM /mg protein, 11.7 ±3.25 nmol NADH/min/mg protein to 39.8±4 nmol NADH/min/mg protein and 0.046 ±0.01 nmol NADPH/min/mg protein. Incubation of both NAC and OTC (0.1mM and 1mM) augmented the GSH level to 0.962 ±0.7 μM/mg protein and g -GCS activity to 112.28 ± 11.85 nmol NADH/min/mg protein , but, GSSGR activity was not significantly increased. The viability of induced cell compared to normal in the presence of 180 m M of HD was significant (P< 0.001), ( 47% to 73%), which was 1.55 times greater in viability. Discussion : These observations indicate that the NAC not only act as a precursor for GSH but also increased GSSGR activity which converted GSSG to GSH . On the other hand ,OTC increased the rate-limiting enzyme( g -GCS) in GSH biosynthesis pathway which increased the GSH levels without inhibition of g -GCS . It is probabely that OTC induces g -GCS expression.
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