ABSTRACT
Resveratrol was irradiated at various doses of 15, 30, 50, and 70kGy for the development of physiological functionalities through modification of the structural properties. Gamma irradiation induced a decrease in the resveratrol peak, and the appearance of several new peaks by gamma irradiation was gradually increased up to 70kGy. Gamma-irradiated resveratrol did not exert cytotoxicity to macrophages in dose ranges from 15 to 70kGy; therefore, 70kGy gamma-irradiated resveratrol was used as the maximum dose throughout subsequent experiments. Treatment of LPS-stimulated macrophages with 70kGy gamma-irradiated resveratrol resulted in a dose-dependent decrease in iNOS-mediated NO, PGE2, and pro-inflammatory cytokine level, such as TNF-α, IL-6 and IL-1ß. 70kGy gamma-irradiated resveratrol significantly inhibited cyclooxygenase-2 levels, as well as the expression of cell surface molecules, such as CD80 and CD86, in LPS-induced macrophages. Furthermore, the inhibitory action of these pro-inflammatory mediators occurred through an inhibition of MAPKs (ERK1/2, p38 and JNK) and NF-κB signaling pathways based on a toll-like receptor 4 in macrophages, which may be closely mediated with the radiolysis products of resveratrol transformed by gamma-irradiation. From these findings, it seems likely that gamma irradiation can be an effective tool for a reduction of the toxicity and play a potent role in the treatment of inflammatory disease.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents/radiation effects , Gamma Rays , Macrophages/drug effects , Stilbenes/pharmacology , Stilbenes/radiation effects , Animals , Anti-Inflammatory Agents/chemistry , B7-1 Antigen/metabolism , B7-2 Antigen/metabolism , Cell Line , Cell Proliferation/drug effects , Cells, Cultured , Dinoprostone/metabolism , Female , Interleukin-1beta/metabolism , Interleukin-6/metabolism , Lipopolysaccharides , Macrophages/metabolism , Mice, Inbred C57BL , Mitogen-Activated Protein Kinases/metabolism , Molecular Structure , NF-kappa B/metabolism , Nitric Oxide/metabolism , Resveratrol , Signal Transduction/drug effects , Stilbenes/chemistry , Toll-Like Receptor 4/metabolism , Tumor Necrosis Factor-alpha/metabolismABSTRACT
BACKGROUND: Green tea polyphenol epigallocatechin-3-gallate (EGCG) has the potential to impact a variety of inflammation-related diseases; however, the anti-inflammatory action of EGCG in endothelial cells has not been understood. Recently, we demonstrated that the 67-kDa laminin receptor (67LR) acts as a cell-surface EGCG receptor. AIM: This research was carried out to clarify the molecular basis for the down-regulation of toll-like receptor 4 (TLR4) signal transduction by EGCG in lipopolysaccharide (LPS)-stimulated endothelial cells. RESULTS: RNAi-mediated silencing of 67LR resulted in an abrogation of the inhibitory action of EGCG on the LPS-induced activation of downstream signaling pathways. Also, we found that EGCG induced a rapid upregulation of Toll-interacting protein (Tollip), a negative regulator of TLR signaling, through 67LR in endothelial cells. RNAi-mediated silencing of Tollip impaired the TLR4 signaling inhibitory activity of EGCG. Additionally, silencing of Tollip resulted in an abrogation of the inhibitory action of EGCG on the LPS-induced expressions of cell-associated adhesion molecules, such as ICAM-1 and VCAM-1. CONCLUSION: Taken together, these novel findings provide new insights into an understanding of negative regulatory mechanisms of the TLR4 signaling pathway and effective therapeutic intervention for the treatment of inflammatory disease.
Subject(s)
Catechin/analogs & derivatives , Endothelial Cells/metabolism , Intracellular Signaling Peptides and Proteins/metabolism , Receptors, Laminin/metabolism , Signal Transduction/drug effects , Toll-Like Receptor 4/metabolism , Animals , Catechin/pharmacology , Cell Adhesion Molecules/genetics , Cell Adhesion Molecules/metabolism , Gene Expression Regulation/drug effects , Humans , Intracellular Signaling Peptides and Proteins/genetics , Lipopolysaccharides/pharmacology , Mice , Mitogen-Activated Protein Kinases/metabolism , NF-kappa B/metabolism , RNA Interference , RNA, Small Interfering/genetics , Receptors, Laminin/genetics , Toll-Like Receptor 4/geneticsABSTRACT
Policosanols, long-chain alcohols, have many beneficial physiological activities. Contents and compositions in perilla seeds (Perilla frutescens) produced in Korea and China were determined. Waxy materials were extracted from perilla seeds using hot hexane. Yield of the waxy materials from perilla seeds was 72.1 mg/100 g of dry weight. Contents and compositions of the waxy materials and policosanols were identified and quantified by TLC, HPLC, and GC. Major components of the waxy materials from Korean and Chinese perilla seeds were policosanols (25.5 and 34.8%, respectively), hydrocarbons (18.8 and 10.5%), wax esters, steryl esters and aldehydes (53.0 and 49.8%), acids (1.7 and 2.1%), and triacylglycerols (1.0 and 2.9%), determined by HPLC. For comparison, waxy materials of sesame seeds were also analyzed. Yield of the waxy materials from sesame seeds were 8.6 mg/100 g. Less than 5% policosanols were detected in the waxy materials extracted from sesame seeds produced in Korea and China. Wax esters or steryl esters accounted for 93-95% of the sesame waxy materials. Policosanols in the perilla seeds were composed of 67-68% octacosanol, 16-17% hexacosanol, 6-9% triacontanol, and others.
