Figure 6

Effects of the scavenger NAC on both oxidative stress and apoptosis of H9c2cells. A) FACS analysis after double labelling with PI and FITC-Annexin V of H9c2 cells treated with 5-FU combined with LF or 200 μμ H₂O₂ or NAC alone or in combination for 48 h. The experiments were performed at least three times and the results were always similar. The results show the % of apoptotic cells derived from the sum of the events calculated as late and early apoptotic cells. Bars, SEs. CTR, untreated cells; H₂O₂, cells treated with 200μM H₂O₂ alone; NAC, cells treated with 5 mM NAC alone; NAC + H₂O_2, cells treated with 5 mM NAC +200μM H₂O₂; 5-FU + LF, cells treated with 5-FU in combination with LF; 5-FU + LF + NAC, cells treated with 5-FU in combination with LF and 5 mM NAC. B) H9c2 were incubated with dihydroethidine and analyzed by flow cytometry as described in “Materials and Methods”. Flow cytometric analysis of H9c2 cells treated with 5-FU combined with LF or 200 μM H₂O₂ or NAC alone or in combination exposed to dihydroethidine used as a probe for measurement of O₂⁻. Representation of the ROS levels expressed as the percentage of mean fluorescence intensity (MFI) derived by dihydroethidine oxidation of H9c2 cells. The experiments were repeated at least three times and gave always similar results. Bars, SDs. CTR, untreated cells; H₂O₂, cells treated with 200μM H₂O₂ alone; NAC, cells treated with 5 mM NAC alone; NAC + H₂O_2, cells treated with 5 mM NAC +200μM H₂O₂; 5-FU + LF, cells treated with 5-FU in combination with LF; 5-FU + LF + NAC, cells treated with 5-FU in combination with LF and 5 mM NAC.