Trx-1 over-expression attenuates oxidative stress injury in MPP+ -induced PC12 cells by regulating NF-κB signaling pathway

AIM:To investigate the inhibitory effect of thioredoxin 1 (Trx-1) over-expression on oxidative stress injury in 1-methyl-4-phenylpyridinium (MPP⁺)-induced rat pheochromocytoma PC12 cells by regulating NF-κB signaling pathway.METHODS:The PC12 cells were damaged by treatment with MPP⁺ at 1, 3 and 5 mmol/L, and the optimal concentration of 3 mmol/L was selected. The cell viability was measured by MTT assay. The oxidative stress indexes lactate dehydrogenase (LDH) activity, superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in the cell culture supernatant were detected, and the protein expression of Trx-1 was determined by Western blot. Lentiviral infection with Ad-Trx-1-GFP sequence was used to establish a model of MPP⁺-treated PC12 cells with Trx-1 over-expression. The effects of Trx-1 over-expression on the cell viability, oxidative stress responses and NF-κB signaling pathway were determined by MTT assay, commercial kits and Western blot. The effects of phorbol 12-myristate 13-acetate (PMA), an activator of NF-κB signaling pathway, on the viability and oxidative stress of PC12 cells were observed. The NF-κB signaling pathway inhibitor pyrrolidine dithiocarbamate (PDTC) was used in MPP⁺-treated PC12 cells with Trx-1 over-expression, and the cell viability and oxidative stress responses were measured. RESULTS:The viability of PC12 cells, SOD activity in the supernatant and protein expression of Trx-1 were decreased, while LDH activity and MDA content in the supernatant were increased significantly by treatment with MPP⁺ at 1, 3 and 5 mmol/L. The effect of MPP⁺ at 3 mmol/L and 5 mmol/L was significantly greater than that at 1 mmol/L (P<0.05), and no significant difference between 3 mmol/L and 5 mmol/L was observed (P>0.05). The inhibitory effect of MPP⁺ on the viability of PC12 cells, and the oxidative stress injury and activation of NF-κB signaling pathway induced by MPP⁺ were significantly attenuated by over-expression of Trx-1. The inhibitory effect of MPP⁺ on the viability of PC12 cells and the oxidative stress injury induced by MPP⁺ were promoted by the activation of NF-κB signaling pathway, while the protective effects of Trx-1 over-expression on the MPP⁺-treated PC12 cells were enhanced by the inhibition of NF-κB signaling pathway. CONCLUSION:Over-expression of Trx-1 protects MPP⁺-treated PC12 cells from oxidative stress injury by regulating NF-κB signaling pathway.