Research Article

Andrographolide Activates Keap1/Nrf2/ARE/HO-1 Pathway in HT22 Cells and Suppresses Microglial Activation by Aβ42 through Nrf2-Related Inflammatory Response

Figure 2

Effect of andrographolide on the expressions and locations of Nrf2 and Keap1. The abbreviation of andrographolide, ANDRO, was used. (a) Total Nrf2 expression was determined by Western blot analysis. The cells were treated with 1, 5, and 10 μM andrographolide for 24 h and the cell lysates were subjected to Western blot analysis. The data are presented as the means SE of triplicates. The significance presented as compared with control group. (b) Nuclear Nrf2 expression was determined by Western blot analysis. Lamin B was used as a nuclear loading control. The data are presented as the means SE of triplicates. The significance presented as compared with control group. (c) The intracellular location and deposition of Nrf2 and Keap1 were examined using immunocytochemistry. The cells were plated and incubated with 10 μM andrographolide for 24 h. Cells were immediately fixed with 3.7% formalin solution and subjected to the next steps described in the “Materials and Methods” section. (d) 3D molecular docking simulation of andrographolide to BTB domain of Keap1 was monitored in silico. The Keap1 protein was visualized by interpolated charge. (e) 2D diagram for non-covalent interactions between andrographolide and wild type or C151W mutated Keap1 was presented.
(a)
(b)
(c)
(d)
(e)