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

Figure 6

HE inhibits TNF-α-induced ROS production in EA.hy926 cells. Cells were preincubated with or without HE (50–200 μg/mL) for 2 h and then stimulated with TNF-α (10 ng/mL) for 15 min. (a) Intracellular ROS levels were measured using a DCFH₂-DA fluorescence microscope (200x magnification). The nonfluorescent cell membrane-permeable probe, DCFH₂-DA, was added to the culture medium at a final concentration of 10 μM. DCFH₂ reacts with cellular ROS and is metabolized into fluorescent DCF. (b) The fluorescence intensity of DCF-stained cells was quantified as a percentage, using Olympus soft image solution software. 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.

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