Fig. 3.

Effect of H₂O₂ and PKC inhibitor treatment on GSH/GSSG levels and H₂O₂ consumption in primary hepatocytes. A: H₂O₂ (300 μM) consumption by hepatocytes in the absence and presence of Ro (10 μM, 1-h pretreatment). ◊, H₂O₂ in medium without hepatocytes; ⧫, H₂O₂ in medium containing hepatocytes; ▪, H₂O₂ in medium containing hepatocytes pretreated with Ro. At the indicated times, media were removed, and H₂O₂ consumption was determined using an oxygen electrode following catalase treatment. B: effect of H₂O₂ (300 μM) on the GSH-to-GSSG ratio (GSH/GSSG ratio). ⧫, H₂O₂ treatment alone; ▪, Ro (10 μM) + H₂O₂. Primary cultured hepatocytes were treated with various doses of H₂O₂ with or without Ro. GSH and GSSG levels were analyzed in the supernatant using HPLC with electrochemical detection as described in materials and methods.