Hericium erinaceus Inhibits TNF-α-Induced Angiogenesis and ROS Generation through Suppression of MMP-9/NF-κB Signaling and Activation of Nrf2-Mediated Antioxidant Genes in Human EA.hy926 Endothelial Cells
HE inhibits TNF-α-induced migration, invasion, and capillary-like tube formation in EA.hy926 cells. (a) Cells were pretreated with HE (50–200 μg/mL) for 2 h. Subsequently, cells were scratched and then stimulated with or without TNF-α (10 ng/mL) for 24 h. Migration was observed using an optic microscope, at a 200x magnification, and the closure of area was calculated using commercially available software. (b) Cells were pretreated with HE (50–200 μg/mL) for 2 h followed by incubation with or without TNF-α (10 ng/mL) for 12 h. Photomicrographs of cells invading under the membrane. The inhibitory percentage of invading cells was quantified and fold change was presented by considering untreated cells (control) as 1-fold. Invasiveness was determined by counting cells in three microscopic fields per sample. (c) Cells were pretreated with HE (50–200 μg/mL) for 2 h and then collected and replaced on Matrigel-coated plates at a density of 1 × 105 cells/well and incubated in the absence or presence of TNF-α (10 ng/mL). After 4 h, the tube formation was determined using a phase-contrast microscope at 200x magnification. The capillary networks were photographed, and the number of tubes was quantified from three random fields. Results are presented as mean ± SD of three independent assays; indicates significant difference compared to control, and indicates significant difference compared to TNF-α alone treatment.