Research Article
Novel Synthetic Coumarin-Chalcone Derivative (E)-3-(3-(4-(Dimethylamino)Phenyl)Acryloyl)-4-Hydroxy-2H-Chromen-2-One Activates CREB-Mediated Neuroprotection in Aβ and Tau Cell Models of Alzheimer’s Disease
Figure 4
Neuroprotective effects of coumarin-chalcone derivatives on ΔK280 tauRD-DsRed-expressing SH-SY5Y cells. (a) Experimental flow chart. On day 1, cells were plated, with retinoic acid (RA, 10 μM) being added to the culture medium. On day 2, Congo red, LM-016, LM-021, or LM-022 (2.5–10 μM) were added to the cells for 8 h, followed by treatment with doxycycline (Dox, 2 μg/ml) to induce ΔK280 tauRD-DsRed expression for 6 days. On day 8, ΔK280 tauRD-DsRed fluorescence, ROS (DCF staining), neurite outgrowth (TUBB3 staining), and caspase 1/AChE activities were measured. (b) Analysis of DsRed fluorescence with Congo red, LM-016, LM-021, or LM-022 (2.5–10 μM) treatment (; two-tailed Student’s -test; and ). The cell numbers counted in each treatment are displayed in the following. The DsRed fluorescence/cell number of untreated cells (Untr) was set as 100% for normalization. (c) ROS assay with Congo red, LM-016, LM-021, or LM-022 (10 μM) treatment (). The cell numbers counted in each treatment are displayed in the following. The relative ROS/cell number of uninduced cells (Dox-) was normalized (100%). (d) TauRD-DsRed RNA, (e) caspase 1, and (f) AChE activity assays with Congo red, LM-016, LM-021, or LM-022 (10 μM) treatment (). For normalization, the caspase 1/AChE activities of uninduced cells (Dox-) were set as 100%. (g) Neurite outgrowth (length, process, and branch) assay with Congo red, LM-016, LM-021, or LM-022 (10 μM) treatment (). (c–g) values: induced (Dox+) vs. uninduced (Dox-) cells (# and ##), or compound-treated vs. untreated (Dox+) cells (, , and ) (one-way ANOVA with a post hoc Tukey test).
| (a) |
| (b) |
| (c) |
| (d) |
| (e) |
| (f) |
| (g) |