Fig. 5.
Intracellular ROS generation induced by 300 μM bupivacaine was measured by labeling with the DCFH-DA dye in pure cultures of astrocytes (A, blue circles; n = 15), and a 5-min incubation with 1 μM cyclosporin A reversed this effect (A, green circles; n = 22); datasets are shown in (B). The effects of 0.3-300 μM bupivacaine on pure cultures of astrocytes were measured by labeling with the DCFH-DA dye (C). The effect of cyclosporin A on reversing bupivacaine-induced ROS generation was reconfirmed using an intracellular ROS kit (D). Data are expressed as the means ± SD. **P < 0.01 compared with the bupivacaine group using an unpaired t test (B), **P < 0.01 compared with the value before treatment with bupivacaine using one-way ANOVA followed by Dunnett’s test (C, n = 23, 21, 18, 22, and 15 for 0, 0.3, 3, 30, and 300 μM bupivacaine, respectively), and **P < 0.01 compared with the bupivacaine group using a two-way ANOVA followed by Bonferroni post hoc test (D, n = 46, 52, 52, 52, and 51 for 0, 0.3, 3, 30, and 300 μM bupivacaine; n = 44, 21, 52, 23, and 24 for 0, 0.3, 3, 30, and 300 μM bupivacaine plus cyclosporin A, respectively). ROS = reactive oxygen species.