Inhibition of TNF-alpha, IL-1 alpha, and IL-1 beta by Pretreatment of Human Monocyte-Derived Macrophages with Menaquinone-7 and Cell Activation with TLR Agonists In Vitro

摘要

Circulatory markers of low-grade inflammation such as tumor necrosis factor-alpha (TNF-alpha), interleukin-1 alpha (IL-1 alpha), and interleukin-1 beta (IL-1 beta) positively correlate with endothelial damage, atheroma formation, cardiovascular disease, and aging. The natural vitamin K2-menaquinone-7 (MK-7) added to the cell culture of human monocyte-derived macrophages (hMDMs) at the same time as toll-like receptor (TLR) agonists did not influence the production of TNF-alpha. When the cells were pretreated up to 6 h with MK-7 before treatment with TLR agonists, MK-7 did not inhibit significantly the production of TNF-alpha after the TLR activation. However, 30 h pretreatment of hMDMs with at least 10 mu M of MK-7 effectively and dose dependently inhibited the proinflammatory function of hMDMs. Pretreatment of hMDMs with 10 lM of MK-7 for 30 h resulted in 20 inhibition of TNF-alpha production after lipopolysaccharide (LPS) activation (P < .05) and 43 inhibition after macrophage-activating lipopeptide (MALP) activation (P < .001). Pathogen-associated molecular pattern (PMPP) activation was inhibited by 20 with MK-7 pretreatment; however, this inhibition was not statistically significant. The 30 h pretreatment of a THP-1-differentiated monocyte cell line with MK-7 resulted in a dose-dependent downregulation of TNF alpha, IL-1 alpha, and IL-1 beta gene expression as evaluated by RNA semiquantitative reverse transcription polymerase chain reaction (RT-PCR). MK-7 is able to modulate immune and inflammatory reactions in the dose-response inhibition of TNF-alpha, IL-1 alpha, and IL-1 beta gene expression and protein production by the healthy hMDMs in vitro.