ABSTRACT
OBJECTIVE: This study assessed anti-inflammatory and antioxidant activities of E. foetidum leaf extract on LPS-activated murine macrophages. METHODS: RAW264.7 cells were pretreated with or without E. foetidum extract for 1 h prior to incubation with LPS for 24 h. Anti-inflammatory activity was evaluated with reference to iNOS, COX-2, TNF-α and IL-6 gene expression. In addition, NO and intracellular ROS generation were determined by Griess method and fluorescence intensity and activation of MAPKs and IκB by Western blotting. RESULTS: Prior treatment with E. foetidum leaf extract inhibited elevation of IL-6, TNF-α, iNOS and COX-2, together with their cognate mRNAs in a dose-dependent manner. NO and intracellular ROS contents were similarly reduced. These effects were due to inhibition of LPS-induced phosphorylation of JNK and p38 as well as IκB. E. foetidum ethanol extract was shown to contain lutein, ß-carotenes, chlorogenic acid, kaempferol and caffeic acid, compounds known to exert these bioactive properties. CONCLUSIONS: E. foetidum leaf extract possesses suppressive effects against pro-inflammatory mediators. Thus, E. foetidum has a high potential to be used as a food supplement to reduce risk of cancer associated with inflammation.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Eryngium/chemistry , Inflammation Mediators/antagonists & inhibitors , Inflammation/drug therapy , Lipopolysaccharides/toxicity , Macrophages/drug effects , Plant Extracts/pharmacology , Animals , Antioxidants/pharmacology , Apoptosis , Blotting, Western , Cell Proliferation , Cells, Cultured , Cyclooxygenase 2/genetics , Cyclooxygenase 2/metabolism , Enzyme-Linked Immunosorbent Assay , I-kappa B Proteins/genetics , I-kappa B Proteins/metabolism , Inflammation/chemically induced , Interleukin-6/genetics , Interleukin-6/immunology , Interleukin-6/metabolism , Macrophages/cytology , Macrophages/immunology , Mice , Mitogen-Activated Protein Kinases/genetics , Mitogen-Activated Protein Kinases/metabolism , NF-KappaB Inhibitor alpha , NF-kappa B/antagonists & inhibitors , NF-kappa B/immunology , NF-kappa B/metabolism , Nitric Oxide/metabolism , Nitric Oxide Synthase Type II/antagonists & inhibitors , Nitric Oxide Synthase Type II/genetics , Nitric Oxide Synthase Type II/metabolism , Phosphorylation/drug effects , Plant Leaves/chemistry , RNA, Messenger/genetics , Reactive Oxygen Species/metabolism , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Signal Transduction/drug effects , Tumor Necrosis Factor-alpha/antagonists & inhibitors , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Pro-inflammatory mediators produced during inflammatory response have been demonstrated to initiate and aggravate pathological development of several chronic diseases. Plant bioactive constituents have been reported to exert anti-inflammatory activities. Various parts of Moringa oleifera have long been used as habitual diets and traditional remedy along the tropical region. Anti-inflammatory activity of boiled M. oleifera pod extract was assessed by measuring pro-inflammatory mediator expression in the lipopolysaccharide-induced murine RAW264.7 macrophage cells. Prior treatment with 31-250 µg/mL M. oleifera extract for 1 h inhibited elevation of mRNA and protein level of interleukine-6, tumor necrosis factor-alpha, inducible nitric oxide synthase, and cyclooxygenease-2, induced by lipopolysaccharide for 24 h in a dose-dependent manner. The suppressive effect was mediated partly by inhibiting phosphorylation of inhibitor kappa B protein and mitogen-activated protein kinases. These results indicate that the anti-inflammatory activity from bioactive compounds present in the M. oleifera pod constituents may contribute to ameliorate the pathogenesis of inflammatory-associated chronic diseases.
Subject(s)
Inflammation Mediators/metabolism , Macrophages/metabolism , Moringa oleifera , Plant Extracts/pharmacology , Animals , Cell Line , Cyclooxygenase 2/biosynthesis , Interleukin-6/biosynthesis , Lipopolysaccharides/immunology , Macrophages/drug effects , Macrophages/immunology , Mice , Mitogen-Activated Protein Kinases/metabolism , Nitric Oxide Synthase Type II/biosynthesis , Nitric Oxide Synthase Type II/metabolism , Phosphorylation/drug effects , RNA, Messenger/genetics , RNA, Messenger/metabolism , Tumor Necrosis Factor-alpha/biosynthesis , Tumor Necrosis Factor-alpha/metabolismABSTRACT
OBJECTIVE: This study investigated anti-inflammatory and antioxidant activities of an ethanol extract from Thai red curry paste. METHODS: The RAW264.7 murine macrophage cell line was incubated with the extract (65-260 µg/mL) with or without lipopolysaccharide. The anti-inflammatory activities of the extract were examined by measuring inducible nitric oxide synthase, cyclo-oxygenase-2, tumor necrosis factor-α, and interleukin-6 mRNA and protein level by reverse transcription-polymerase chain reaction, western blotting, and enzyme-linked immunosorbent assay, respectively. Nitric oxide production and intracellular reactive oxygen species generation were determined by the Griess method and fluorescence intensity. The activation of mitogen-activated protein kinases and inhibitor κB were determined by western blot. RESULTS: Exposure of cells with the extract significantly suppressed lipopolysaccharide-induced nitric oxide production and inducible nitric oxide synthase, cyclo-oxygenase-2, tumor necrosis factor-α, and interleukin-6 expressions (P < 0.05) by dose-dependently without cytotoxic effect. Intracellular reactive oxygen species significantly decreased (P < 0.05) in lipopolysaccharide-induced RAW264.7 cells. The inhibitory effect was mediated partly by inhibiting activation of inhibitor κB-α and mitogen-activated protein kinases. CONCLUSION: These results suggest that the anti-inflammatory and antioxidant properties of Thai red curry paste stem from bioactive compounds present in the spice and herb constituents. The health benefits of Thai red curry paste warrant further investigations in vivo.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Antioxidants/pharmacology , Cytokines/metabolism , Inflammation Mediators/metabolism , Macrophages/drug effects , Plant Extracts/pharmacology , Animals , Cell Line , Cyclooxygenase 2 Inhibitors/pharmacology , Dose-Response Relationship, Drug , Interleukin-6/metabolism , Lipopolysaccharides , Macrophages/metabolism , Mice , Mitogen-Activated Protein Kinases/metabolism , Nitric Oxide/biosynthesis , Nitric Oxide Synthase Type II/metabolism , Phytotherapy , Reactive Oxygen Species/metabolism , Tumor Necrosis Factor-alpha/metabolismABSTRACT
Nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha) play important roles in inflammatory processes. This study examined whether 13 spices/herbs commonly used in Thai dishes modulate the production of NO and TNF-alpha by the RAW 264.7 mouse macrophage cell line pretreated with plant extracts (1-100 microg/mL) prior to activation by bacterial lipopolysaccharide (LPS). Tested plant tissues were extracted with ethanol with the exception of roselle, which was extracted with 70% acetone. Eight of the 13 plant extracts inhibited NO and TNF-alpha production in a dose-dependent manner without exerting cytotoxicity. Extract from Limnophila aromatica (Kyeng) was the most robust suppressor of NO production, followed by dill, kaffer lime, chili, Teaw, mint, sweet basil, and pea eggplant, respectively (range of 50% inhibitory concentration [IC(50)] = 11.4-74.6 microg/mL). Kyeng also exhibited the greatest inhibition of TNF-alpha production (IC(50) = 10.5 microg/mL). IC(50) values for NO and TNF-alpha production in LPS-activated RAW 264.7 cells for these extracts were highly correlated (r = 0.772, P = .025). These results suggest that extracts from some spices/herbs in the habitual Thai diet possess anti-inflammatory activity. Moreover, the results support the use of NO production in LPS-activated RAW 264.7 cells as a rapid and cost-effective tool for screening the anti-inflammatory activity of extracts of spices/herbs.
