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Macrophages are essential for regulating the physiology of pregnancy; however, excessive inflammatory responses to macrophages, induced by infection and/or endogenous danger signals, may potentially result in complications during pregnancy. Advanced glycation end-products (AGE) and lipopolysaccharides (LPS) are known to induce inflammation and are associated with adverse developmental outcomes. The aim of the present study was to examine the effect of AGE and LPS on cytokines in the J774 murine macrophage cell line and the potential effect of resveratrol on AGE- and LPS-induced inflammation in macrophages. AGE and LPS significantly increased IL-6 mRNA expression and secretion in J774 macrophages (P&lt;0.05). Although AGE and LPS significantly stimulated IL-1β mRNA expression (P&lt;0.05), they had no significant effect on IL-1β secretion. To assess the receptors for AGE and LPS, including receptor for AGE (RAGE) and Toll-like receptor (TLR4), blocking reagents (RAGE antagonist or TLR4 inhibitor) were added to the J774 macrophages. IL-6 secretion induced by AGE or LPS was significantly inhibited by pretreatment with RAGE antagonist (P&lt;0.05) or TLR4 inhibitor (P&lt;0.05). IL-6 secretion was dependent on nuclear factor (NF)-κB activation and the production of reactive oxygen species (ROS; P&lt;0.05). Resveratrol suppressed mRNA expression and intracellular IL-6 production, resulting in significantly decreased IL-6 secretion after treatment with LPS or AGE (P&lt;0.01). Furthermore, treatment with Ex527, which is a sirtuin-1 (SIRT1) inhibitor, significantly attenuated the anti-inflammatory effect of resveratrol (P&lt;0.05), and treatment with 5-aminoimidazole-4-carboxamide ribonucleotide, which is a 5' adenosine monophosphate-activated protein kinase (AMPK) activator, resulted in a significant decrease in IL-6 secretion in J774 macrophages (P&lt;0.05). The results of the present study indicated that AGE and LPS increase IL-6 secretion depending on NF-κB activation and ROS production through RAGE and/or TLR4 in the J774 murine macrophage cell line. Based on the present study, resveratrol appears to be an effective regulator of the inflammatory responses associated with SIRT1 and AMPK activation in macrophages. These results suggest that resveratrol may have therapeutic applications for controlling immune responses during pregnancy.

experimental and therapeutic medicine

Advanced glycation end products and lipopolysaccharides stimulate interleukin‑6 secretion via the RAGE/TLR4‑NF‑κB‑ROS pathways and resveratrol attenuates these inflammatory responses in mouse macrophages