Research Paper Volume 11, Issue 18 pp 7961—7977

Kinsenoside ameliorates intervertebral disc degeneration through the activation of AKT-ERK1/2-Nrf2 signaling pathway

Figure 3. Kin activates the AKT-ERK1/2-Nrf2 signaling pathway in NPCs. (A) Real-time PCR assay of Nrf2, SOD2 and NQO1 in the NPCs treated with Kin (25 μg/ml) for indicated time points (0, 2, 4, 8, 16 or 24 h) or different concentrations of Kin (0, 5, 10 or 25 μg/ml) for 24 h. (B) The western blotting of Nrf2, SOD2 and NQO1 in the NPCs as treated above. (C) Nuclear translocation of Nrf2 in the NPCs treated with Kin (25 μg/ml) for 8 h was observed by immunofluorescence staining. (D, E) The western blotting and quantitative protein levels of p-AKT, AKT, p-ERK1/2, ERK1/2, p-p38, p38, p-JNK and JNK in the NPCs treated with Kin (25 μg/ml) for indicated time points (0, 15, 30, 60, 90 or 120 min) or different concentrations of Kin (0, 5, 10 or 25 μg/ml) for 120 min. (F, G) The western blotting and quantitative protein levels of p-AKT, AKT, p-ERK1/2, ERK1/2, nuclear Nrf2, SOD2 and NQO1 in the NPCs pretreated with U0126 (10 μM) or LY294002 (10 μM) for 1 h prior to incubation with Kin (25 μg/ml). All data are expressed as mean ± SD of at least three independent experiments.