Journal: Oxidative Medicine and Cellular Longevity

Article Title: Ethanol Extract of Centipeda minima Exerts Antioxidant and Neuroprotective Effects via Activation of the Nrf2 Signaling Pathway

doi: 10.1155/2019/9421037

Figure Lengend Snippet: ECM inhibits oxidative stress-induced cell death. (a) Relative viability of SH-SY5Y and PC12 cells treated with 0.5-2 μ g/ml ECM at 37°C for 24 h. (b) Relative viability of SH-SY5Y and PC12 cells pretreated with 0.5-2 μ g/ml ECM for 2 h and then exposed to 300 μ M tBHP for an additional 6 h. (c) Relative viability of SH-SY5Y and PC12 cells pretreated with 0.5-2 μ g/ml ECM for 2 h and then exposed to 10 mM glutamate for an additional 24 h. Cells were pretreated with ECM and then treated with 300 μ M tBHP for 4 h (d) or 10 mM glutamate for 18 h (e). The morphological changes were observed in the bright field of microscope. Scale bar, 50 μ m. All data are normalized to control cells and presented as the mean ± SEM of three independent experiments. # p < 0.05 and ## p < 0.01 in comparison with control cells. ∗ p < 0.05 and ∗∗ p < 0.01 in comparison with the cells exposed to glutamate (b) or tBHP (c) alone.

Article Snippet: The human neuroblastoma cell line SH-SY5Y was purchased from the cell bank Interlab Cell Line Collection (Genova, Italy).

Techniques: Microscopy, Control, Comparison

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Ethanol Extract of Centipeda minima Exerts Antioxidant and Neuroprotective Effects via Activation of the Nrf2 Signaling Pathway

doi: 10.1155/2019/9421037

Figure Lengend Snippet: ECM attenuates oxidative stress-induced mitochondrial dysfunction. The cells were pretreated with 0.5-2 μ g/ml ECM and 50 μ M vitamin E for 2 h and then exposed to 300 μ M tBHP for an additional 6 h. (a) SH-SY5Y (upper panel) or PC12 cells (lower panel) were treated with DCFH-DA for 30 min; Hoechst 33342 was used to counterstain cell nuclei. Scale bar, 50 μ m. (b) The percentage of ROS-positive cells among cultured SH-SY5Y or PC12 cells was quantified and was shown as histogram. Intracellular MDA content (c), SOD activity (d), and GSH levels (e) were detected using a kit assay and are presented as a histogram. (f) SH-SY5Y cells were pretreated with 0.5-2 μ g/ml ECM for 2 h and then exposed to 300 μ M tBHP for an additional 6 h. The mitochondrial membrane potential was determined using the JC-1 fluorescence probe, and representative pictures have been shown for comparison. (g) Western blot analysis was performed using antibodies against Bax and Bcl-2, and β -actin was used as a loading control. (h) The ratio of Bcl-2 to Bax was quantified by densitometry and is shown as a histogram. The results are shown as the mean ± SEM of three independent experiments. ## p < 0.01 in comparison with control cells. ∗ p < 0.05 and ∗∗ p < 0.01 in comparison with the cells exposed to tBHP alone.

Article Snippet: The human neuroblastoma cell line SH-SY5Y was purchased from the cell bank Interlab Cell Line Collection (Genova, Italy).

Techniques: Cell Culture, Activity Assay, Membrane, Fluorescence, Comparison, Western Blot, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Ethanol Extract of Centipeda minima Exerts Antioxidant and Neuroprotective Effects via Activation of the Nrf2 Signaling Pathway

doi: 10.1155/2019/9421037

Figure Lengend Snippet: ECM inhibits oxidative stress via the regulation of the MAPK and Nrf2 signaling pathways. The cells were pretreated with 0.5-2 μ g/ml ECM and 50 μ M vitamin E for 2 h and then exposed to 300 μ M tBHP for an additional 1 h. Western blot analysis was performed using antibodies against p-p38, p38, p-ERK, ERK, p-JNK, and GAPDH; β -actin was used as a loading control in SH-SY5Y (a) and PC12 cells (b). Nuclear and cytoplasmic proteins were extracted after treatment, and Nrf2 was detected using Western blot analysis in SH-SY5Y (c) and PC12 cells (d); GAPDH and Lamin B1 were used as loading controls. Western blot analysis was performed using antibodies against HO-1, NQO1, and SOD2 in SH-SY5Y (e) and PC12 cells (f); GAPDH was used as a loading control.

Article Snippet: The human neuroblastoma cell line SH-SY5Y was purchased from the cell bank Interlab Cell Line Collection (Genova, Italy).

Techniques: Protein-Protein interactions, Western Blot, Control

Journal: Oxidative Medicine and Cellular Longevity

Article Title: Ethanol Extract of Centipeda minima Exerts Antioxidant and Neuroprotective Effects via Activation of the Nrf2 Signaling Pathway

doi: 10.1155/2019/9421037

Figure Lengend Snippet: The active compounds EBSC-26A−EBSC-26D ameliorate oxidative stress. (a) SH-SY5Y and PC12 cells were treated with 0.5-2 μ M EBSC-26A−EBSC-26D at 37°C for 24 h (upper panel) or pretreated with 0.5-2 μ M EBSC-26A−EBSC-26D for 2 h and then exposed to 300 μ M tBHP for an additional 6 h (lower panel); the relative viability of cells was measured by MTT assay. (b, c) SH-SY5Y cells were probed with DCFH-DA (b) and the percentage of ROS-positive cells among culture cells was quantified (c). Scale bar, 50 μ m. (d) After pretreatment with 1-2 μ M EBSC-26A, EBSC-26B, and vitamin E for 2 h, SH-SY5Y cells were exposed to 300 μ M tBHP for an additional 6 h, and then nuclear Nrf2, HO-1, NQO-1, and SOD2 levels were detected by Western blot; β -actin was used as a loading control. (e) Relative protein levels were quantified by densitometry and normalized to Lamin B or β -actin. The results are shown as the mean ± SEM of three independent experiments. # p < 0.05 in comparison with control cells. ∗ p < 0.05 and ∗∗ p < 0.01 in comparison with the cells exposed to tBHP alone.

Article Snippet: The human neuroblastoma cell line SH-SY5Y was purchased from the cell bank Interlab Cell Line Collection (Genova, Italy).

Techniques: MTT Assay, Western Blot, Control, Comparison