ABSTRACT
Objective To evaluate the role of nicotinamide adenine dinucleotide phosphate hydrogen (NADPH) oxidase in bupivacaine-induced injury to neurons.Methods SH-SY5Y cells were seeded in 96-well culture plates at a density of 5× 104 cells/well and randomly divided into 3 groups (n=24each) using a random number table:control group (group C),bupivacaine group (group B),and apocynin (NADPH oxidase inhibitor) + bupivacaine group (group A+B).The cells were cultured in a serum-free medium in group C.The cells were cultured in a serum-free medium containing 1 mmol/L bupivacaine in group B.In group A + B,the cells were cultured for 30 min in a serum-free medium containing apocynin 100 μmol/L,and then cultured in a serum-free medium containing 1 mmol/L bupivacaine.At 2,4 and 6 h of incubation,the cells in 6 wells of each group were selected to evaluate the cell viability by MTS assay.At 4 h of incubation,the cells in 6 wells of each group were selected to detect reactive oxygen species (ROS) level by flow cytometry.Results Compared with group C,the cell viability was significantly decreased at 4 and 6 h of incubation,and the production of ROS was increased at 4 h of incubation in group B (P＜ 0.05).Compared with group B,the cell viability was significantly increased at 4 and 6 h of incubation,and the production of ROS was decreased at 4 h of incubation in group B (P＜0.05).Conclusion NADPH oxidase is involved in bupivacaine-induced injury to neurons.