آلوده به خردل در محيط كشت THF2FF و نقش حفاظتي آن در سلول هاي )OTC( آموتياز وليدين -4- كربوكسيلات 2-L )NAC( استيل سييستيين -N بوسيله )GSH( بررسي اثر تحريكي استامينوفن روي ميزان القاء كلوتاتيون

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

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 Gama-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 tM 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 25miuM acetaminophen increased the GSH,y-GCS and GSSGR activities respectively from 0.331+0.12miuM/mg protein to 0.784+0.11 miuM/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/minlmg protein. Incubation of both NAC and OTC (0.1mM and 1mM) augmented the GSH level to 0.962 +0.7 p.Mlmg protein and Gama-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 180µM 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(Gama-GCS) in GSH biosynthesis pathway which increased the GSH levels without inhibition of y-GCS . It is probabely that OTC induces Gama-GCS expression.