Antioxidants decrease toxin-induced expression of TNF
RAW 264.7 cultures were prepared as described in Materials and Methods and challenged with sublytic amount of LeTx known to induce a large cytokine response (PA = 0.1 µg/ml, LF = 10−6 µg/ml) in the presence of various antioxidants: DTT and BME (10 mM), ethanol and DMSO (10%, v/v), vitamins C and E and mepacrine (100 mM). N-acetyl-L-cysteine (50 mM) was preincubated with Mϕ cells for 16 hr prior to LeTx challenge to allow for incorporation into glutathione. After 6 hr in toxin-containing media, cells were lysed with 0.1% Triton X-100 and total (cellular plus extracellular) TNFα (a) and IL-1β (b) concentrations were determined by ELISA. Experiments were run in duplicate with two experimental series. SEM values for each point were <18% of the indicated value.