Antrodia salmonea inhibits TNF-α-induced angiogenesis and atherogenesis in human endothelial cells through the down-regulation of NF-κB and up-regulation of Nrf2 signaling pathways

Abstract

Ethnopharmacological relevance: Antrodia salmonea (AS) is known as a traditional Chinese medicine, but very few biological activities have been reported.

Materials and methods: The present study was aimed to investigate the anti-angiogenic and anti-atherosclerotic potential of the fermented culture broth of AS against tumor necrosis factor-α (TNF-α)-stimulated human endothelial (EA.hy 926) cells.

Results: The non-cytotoxic concentrations of AS significantly inhibited TNF-α-induced migration/invasion and capillary-like tube formation in EA.hy 926 cells. Furthermore, AS suppressed TNF-α-induced activity and expression of matrix metalloproteinase-9 (MMP-9), and cell-surface expression of intercellular adhesion molecule-1 (ICAM-1), which was associated with abridged adhesion of U937 leukocytes to endothelial cells. Moreover, AS significantly down-regulated TNF-α-induced nuclear translocation and transcriptional activation of nuclear factor κB (NF-κB) followed by suppression of I-κB degradation and phosphorylation of I-κB kinase-α (IKKα). Notably, the protective effect of AS was directly correlated with the increased expression of hemeoxygenase-1 (HO-1) and γ-glutamylcysteine synthetase (γ-GCLC), which was reasoned by nuclear translocation and transactivation of NF-E2 related factor-2 (Nrf2)/antioxidant response element (ARE). Furthermore, HO-1 knockdown by HO-1-specific shRNA diminished the protective effects of AS on TNF-α-stimulated invasion, tube formation, and U937 adhesion in EA.hy 926 cells.

Conclusions: Taken together, these results suggest that Antrodia salmonea may be useful for the prevention of angiogenesis and atherosclerosis.

Keywords: ARE; AS; Antrodia salmonea; DMEM; Dulbecco's Modified Eagle's medium; EA.hy 926 cells; FBS; FITC; GSH; HO-1; Hemeoxygenase-1; I-κB; I-κB kinase-α; ICAM-1; IKKα; MMP-9; NF-κB; Nrf2; PBS; TNF-α; antioxidant response element; fetal bovine serum; fluorescein isothiocyanate; glutathione; inhibitor-κB; intercellular adhesion molecule-1; matrix metalloproteinase-9; nuclear factor erythroid 2-related factor 2; nuclear factor-κB; phosphate buffer saline; tumor necrosis factor-α; γ-GCLC; γ-glutamylcysteine synthetase.