Research Paper Volume 12, Issue 13 pp 13463—13476

Mechanistic study of mtROS-JNK-SOD2 signaling in bupivacaine-induced neuron oxidative stress

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Figure 6. SOD2 gene knock-down enhanced BPV-induced neurotoxic injury and mito-TEMPO attenuated above injury. Con: cells transfected with silencer negative control siRNA; siSOD2: cells transfected with SOD2 siRNA. BPV: cells transfected with silencer negative control siRNA and cultured with 2.0 mM BPV for 60 min; siSOD2+BPV: cells transfected with SOD2 siRNA and cultured with 2.0 mM BPV for 60 min. (A, B) MMP was monitored by determining the relative amount of dual emissions from mitochondrial JC-1 monomers or aggregates using flow cytometry. Mitochondrial depolarization is indicated by a decrease in the polymer/monomer fluorescence. scale bar 200 μm. (C, D) MDA and 8-OHdG were detected by ELISA; (EH) cells apoptosis was detected with flow cytometry and cleaved caspase-9 expression; (I) LDH production in cells treated with 2.0 mM BPV for 60 min and/or pretreated with 25 μM mito-TEMPO for 60 min. Values are the mean± SEM of n = 3, *: P< 0.05.