Effect of Danggui Shaoyaosan Ameliorates in Alleviating H2O2-induced SH-SY5Y Cells Oxidative Stress Through Modulation of NF-κB Pathway / 中国实验方剂学杂志

ABSTRACT

Objective:To investigate the regulatory effect of Danggui Shaoyaosan (DSS)-containing serum on oxidative stress and inflammation in H2O2-induced SH-SY5Y cells. Method:Methyl thiazolyl tetrazolium(MTT) assay was used to determine the cell activity and construct the H2O2-induced cell damage model with the optimal time and dose. Normal group, model group and high, medium and low-dose DSS groups(2.5%, 5%, 10%) were set up. MTT method was used to detect cell activity, spectrophotometry anti-oxidation indexes of malonaldehyde (MDA), catalase (CAT) and superoxide dismutase (SOD) and glutathione (GSH). Real-time quantitative PCR (Real-time PCR) was used to detect tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) mRNA expressions. And immunofluorescence test was adopted to detect nuclear transcription factor-κB(NF-κB) p65 nuclear translocation of the DSS after the intervention. Result:After 24 h intervention with 250 μmol·L-1 H2O2, SH-SY5Y cell viability was about 55%, which was the best modeling condition. After high, medium and low-dose DSS intervention on H2O2-damaged cell model, compared with the model group, the cell activity showed a dose-dependent increase (P<0.05), MDA was significantly reduced (P<0.05), and antioxidant indexes CAT, SOD and GSH were significantly increased (P<0.05). H2O2 could significantly increase SH-SY5Y cell inflammatory factor TNF-α, IL-6 and IL-1β mRNA expressions, and promote activation of cytoplasmic NF-κB and nuclear translocation. DSS-containing serum showed a dose-dependent inhibition of NF-κB p65 from nuclear, and reduced inflammatory factor levels, such as TNF-α, IL-6 and IL-1β. Conclusion:DSS-containing serum can significantly reduce the oxidative damage in H2O2-induced SH-SY5Y cells by improving their antioxidant status, and reduce the inflammatory response by inhibiting NF-κB signaling pathway.