The n-butanol fraction of Naematoloma sublateritium suppresses the inflammatory response through downregulation of NF-κB in human endothelial cells

Abstract

Abstract. Naematoloma sublateritium (Fr.) P. Karst is a chestnut mushroom that is currently a popular edible fungus in the USA, Japan, China and Korea. Although its therapeutic potential in the treatment of diseases has been demonstrated, the pharmacological effect of N. sublateritium (NS) has been poorly studied. In the present study, we demonstrate for the first time that NS suppresses TNF-α-induced inflammatory response in human umbilical vein endothelial cells. The n-butanol fraction of NS (BFNS) inhibited TNF-α-induced monocyte adhesion to endothelial cells in a dose-dependent manner. The anti-adhesive activity of BFNS correlated with suppressed expression of vascular cell adhesion molecule-1, intercellular adhesion molecule-1, monocyte chemoattractant protein-1 and interleukin-8 at both the mRNA and protein levels. In addition, BFNS dose-dependently decreased the expression of inducible nitrogen oxygen synthase (iNOS) and cyclooxygenase-2 (COX-2). Notably, BFNS significantly regulated the nuclear factor (NF)-κB transcriptional activity that was activated by TNF-α stimulation. When considered together, these results suggest that BFNS inhibits the expression of TNF-α-induced adhesion molecules in addition to regulating the iNOS/COX-2 pathways through the modulation of NF-κB in endothelial cells. In conclusion, we propose that BFNS may be a potential therapeutic agent against vascular inflammation, such as atherosclerosis. IntroductionAtherosclerosis, a cardiovascular disease, is a leading cause of death worldwide and is the result of a complicated inflammatory processes on the inner wall of large blood vessels. Immune responses can cause vascular endothelial dysfunctions that eventually disturb blood flow (1). In the early pathogenesis of atherosclerosis, the endothelium is activated by a variety of relevant stimuli, including oxidized lipids, cytokines, or signaling molecules such as endotoxins, and viruses, subsequently expressing specific adhesion receptors that enhance mononuclear cell recruitment into the vessel wall (2,3).The immune reactions in atherosclerotic lesions are mediated by nitric oxide (NO) and cyclooxygenase-2 (COX-2) (4). In the physiological function of the endothelium, NO production is required to inhibit monocyte adhesion and smooth muscle cell chemotaxis and proliferation (5). However, high levels of NO, which are produced by inducible NO synthase (iNOS) in response to stimuli, induce subsequent inflammatory reactions, thus contributing to the progression of atherosclerosis (6). COX-2 has also been associated with pro-atherogenic reactions, generating lipid mediators of inflammation, such as prostaglandin E 2 (PGE 2 ) (7).Epidemiological data continue to support the premise that dietary intake of natural products may improve vascular endothelial dysfunction in both in vivo and in vitro atherosclerotic models (8,9). For example, popular and edible mushrooms have been traditionally used to maintain health, and their components have been used in the prevention and treatment of diseases i...

Anthocyanin-Rich Extract from Red Chinese Cabbage Alleviates Vascular Inflammation in Endothelial Cells and Apo E−/− Mice

“…The effect of ArCC treatment on VCAM-1 or NF-κB transcription was determined using luciferase reporter assay as described previously [ 37 , 38 ]. The HEK 293T/VCAM-1-luc cells were incubated with various concentrations of ArCC in an opaque-walled 96-well plate (4 × 10 4 cells/well) for 1 h, followed by TNF-α stimulation for 6 h. The cells were treated with luciferin for 30 min at 37 °C, and the luminescence was measured by using a luminometer (Glomax, Promega, Madison, WI, USA).…”

“…Cells were treated with the specified concentrations of ArCC and lysed, as previously described [ 38 ]. The cell lysate was cleared by centrifugation at 12,000× g for 20 min at 4 °C, and the supernatant was used for immunoblotting.…”

“…HUVECs were cultured on coverslips, followed by treatment with 100 and 300 μg/mL of ArCC for 6 h and stimulation with TNF-α. After permeabilization with 0.1% Triton X-100, the cells were treated with anti-p65 NF-κB antibody (1:500 dilution; Santa Cruz Biotechnology, Dallas, TX, USA) for 2 h. Immunofluorescence staining was performed as described previously [ 38 ]. The cells were washed and incubated with Alexa Fluor 488-conjugated secondary antibody (1:1000 dilution; Molecular Probes, Eugene, OR, USA) for 1 h. After washing, the cells were counterstained with propidium iodide (PI) for 1 min.…”

Triterpenes from the Mushroom Hypholoma lateritium: Isolation, Structure Determination and Investigation in Bdelloid Rotifer Assays

“…Naematolin, a caryophyllane derivative with cytotoxic activity, was isolated from cultures of H. lateritium [6]. Attempts have been made to characterize the possible mechanism of action behind the antitumor and anti-inflammatory effects of H. lateritium extracts [7,8], as well as to explore the antimicrobial, antioxidant, and xanthine oxidase inhibitory properties of this species [9,10,11]. The current study was performed to identify the major secondary metabolites of H. lateritium and characterize their toxicity in bdelloid rotifer assays.…”