scholarly journals Schisandra Chinensis Lignans Suppresses the Production of Inflammatory Mediators Regulated by NF-κB, AP-1, and IRF3 in Lipopolysaccharide-Stimulated RAW264.7 Cells

Molecules ◽  
2018 ◽  
Vol 23 (12) ◽  
pp. 3319 ◽  
Author(s):  
Gan Luo ◽  
Brian Cheng ◽  
Hui Zhao ◽  
Xiu-Qiong Fu ◽  
Ran Xie ◽  
...  
Keyword(s):  
Inflammatory Mediators ◽  
Nuclear Translocation ◽  
Chinese Herb ◽  
Underlying Mechanism ◽  
Raw264.7 Cells ◽  
Schisandra Chinensis ◽  
Protein Levels ◽  
Cytokines And Chemokines ◽  
Raw264.7 Macrophages ◽  
Anti Inflammatory

Schisandra Fructus (SF) is a traditional Chinese herb used in the treatment of inflammatory disorders like hepatitis. One of the main anti-inflammatory components of SF is the lignans. However, the underlying anti-inflammatory mechanism of Schisandra Chinensis lignans (SCL) remains unclear. This study aims to investigate the effects of SCL on inflammatory mediators in lipopolysaccharide-stimulated RAW264.7 cells and explore the underlying mechanism. The production of nitric oxide (NO) was determined by Griess reaction. ELISA was used to determine cytokine levels and chemokines secretion. To estimate protein levels and enzyme activities, we employed Western blotting. Nuclear localization of NF-κB, AP-1, and IRF3 was detected using immunofluorescence analyses. The results showed that SCL significantly reduced the release of inflammatory mediators, including NO and PGE2, which may be related to down-regulation of iNOS and COX-2 expression. The production of cytokines and chemokines was suppressed by SCL treatment. SCL also decreased the phosphorylation of IKKα/β, IκB-α, Akt, TBK1, ERK, p38, JNK, NF-κB (p65), AP-1 (c-Jun), and IRF3 in RAW264.7 macrophages activated with LPS. The nuclear protein levels and nuclear translocation of AP-1, NF-κB and IRF3 were suppressed by SCL. These results indicated that SCL suppressed the IKKα/β/NF-κB, MAPKs/AP-1 and TBK1/IRF3 signaling pathways in LPS-stimulated RAW264.7 macrophages.

The effect of total flavonoids from Saussurea involucrata on the secretion of pro-inflammatory mediators in lipopolysaccharide stimulated RAW264.7 macrophages

2020 ◽  
Author(s):  
Li-Shan Yan ◽  
Li Wang ◽  
Brian Chi-Yan Cheng ◽  
Yu Ding ◽  
Jing Kong ◽  
...  
Keyword(s):  
Inflammatory Mediators ◽  
Nuclear Translocation ◽  
Cell Model ◽  
Inflammatory Diseases ◽  
Underlying Mechanism ◽  
Total Flavonoids ◽  
Saussurea Involucrata ◽  
Cytokines And Chemokines ◽  
Raw264.7 Macrophages ◽  
Anti Inflammatory

Abstract Background Saussurea involucrate (SI) has long been used to treat inflammatory diseases, such as rheumatoid arthritis. The main active constituents of SI are flavonoids, which are a class of polyphenolic compounds. However, few studies have investigated the anti-inflammatory activity of the total flavonoids of SI (FSI). The mechanism underlying this action is still not fully understood. In the present study, we employed RAW264.7 cell line as an inflammatory cell model to investigate the anti-inflammatory effects of FSI and explore the corresponding molecular mechanisms.Methods We extracted FSI using chromatographic column method. The cell viability was determined by MTT assay. The production of nitric oxide (NO) was detected by Griess assay. The release of cytokines and chemokines were determined by ELISA assays. The nuclear translocation of p65, c-Jun, and IRF3 was detected by immunofluorescence microscopy. Western blotting analysis was performed to determine the related protein expression.Results The results showed that the amount of FSI extracted from SI was 751.5 mg/g. The production of inflammatory mediators was effectively inhibited by FSI. Meanwhile, FSI also suppressed the nuclear translocation of p65, c-Jun, and IRF3. The elevated expression of iNOS, COX-2, p-IKKα/β, p-TBK1, p-IκBα, p-ERK, p-p38, p-JNK, p-p65, p-c-Jun, p-IRF3 induced by LPS was remarkably reduced by FSI treatment.Conclusion These findings indicated that FSI has a potential ability to inhibit the secretion of pro-inflammatory mediators and the underlying mechanism may be related to block the p65, c-Jun, and IRF3 signaling pathways. This study provided evidence for the anti-inflammatory mode and the underlying mechanism of FSI.

scholarly journals Paeonol attenuates inflammation by targeting HMGB1 through upregulating miR-339-5p

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Liyan Mei ◽  
Meihong He ◽  
Chaoying Zhang ◽  
Jifei Miao ◽  
Quan Wen ◽  
...  
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Chinese Herb ◽  
Cecal Ligation ◽  
Underlying Mechanism ◽  
Raw264.7 Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Life Threatening ◽  
Moutan Cortex ◽  
Anti Inflammatory

AbstractSepsis is a life-threatening disease caused by infection. Inflammation is a key pathogenic process in sepsis. Paeonol, an active ingredient in moutan cortex (a Chinese herb), has many pharmacological activities, such as anti-inflammatory and antitumour actions. Previous studies have indicated that paeonol inhibits the expression of HMGB1 and the transcriptional activity of NF-κB. However, its underlying mechanism is still unknown. In this study, microarray assay and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) results confirmed that paeonol could significantly up-regulate the expression of miR-339-5p in RAW264.7 cells stimulated by LPS. Dual-luciferase assays indicated that miR-339-5p interacted with the 3′ untranslated region (3′-UTR) of HMGB1. Western blot, immunofluorescence and enzyme-linked immunosorbent assay (ELISA) analyses indicated that miR-339-5p mimic and siHMGB1 both negatively regulated the expression and secretion of inflammatory cytokines (e.g., HMGB1, IL-1β and TNF-α) in LPS-induced RAW264.7 cells. Studies have confirmed that IKK-β is targeted by miR-339-5p, and we further found that paeonol could inhibit IKK-β expression. Positive mutual feedback between HMGB1 and IKK-β was observed when we silenced HMGB1 or IKK-β. These results indicated that paeonol could attenuate the inflammation mediated by HMGB1 and IKK-β by upregulating miR-339-5p expression. In addition, we constructed CLP model mice by cecal ligation and puncture. Paeonol was used to intervene to investigate its anti-inflammatory effect in vivo. The results showed that paeonol could improve the survival rate of sepsis mice and protect the kidney of sepsis mice.

scholarly journals Methyl 3,4,5-trimethoxycinnamate suppresses inflammation in RAW264.7 macrophages and blocks macrophage–adipocyte interaction

2020 ◽  
Vol 28 (5) ◽  
pp. 1315-1326 ◽  
Author(s):  
Olumayokun A. Olajide ◽  
Idowu S. Akande ◽  
Carlos da Silva Maia Bezerra Filho ◽  
Izabela Lepiarz-Raba ◽  
Damião Pergentino de Sousa
Keyword(s):  
Dna Binding ◽  
Glucose Uptake ◽  
Luciferase Activity ◽  
Antioxidant Activities ◽  
Raw264.7 Cells ◽  
Cytokines And Chemokines ◽  
Raw264.7 Macrophages ◽  
Cox 2 ◽  
Anti Inflammatory ◽  
Griess Assay

Abstract Methyl 3,4,5-trimethoxycinnamate (MTC) is a bioactive natural phenylpropanoid. We evaluated anti-inflammatory effects of synthetic MTC in RAW264.7 macrophages and RAW264.7–3T3-L1 adipocytes co-culture. Levels of cytokines and chemokines, as well as NO and PGE2 in cell supernatants were analysed using ELISAs, Griess assay and enzyme immunoassays, respectively. In-cell cytoblot was used to assess levels of proteins; while DNA binding and reporter gene assays were used to measure transcription factor DNA binding and transcriptional activities, respectively. Glucose uptake in adipocytes was evaluated with 2‐deoxy‐2‐[(7‐nitro‐2, 1, 3‐benzoxadiazol‐4‐yl) amino]‐d‐glucose uptake. MTC (5–20 µM) suppressed LPS + IFNγ-induced release of TNFα, IL-6 and IL-1β, as well as NO/iNOS and PGE2/COX-2 levels in RAW264.7 cells. Furthermore, there was a reduction in phospho-IκB and phospho-p65 proteins, accompanied by a reduction in total IκB in RAW264.7 cells. Further studies showed that MTC also produced a reduction in NF-κB DNA binding and luciferase activity. Treatment of RAW264.7 cells with MTC (5–20 µM) resulted in enhanced DNA binding of Nrf2 and an increase in ARE-luciferase activity. In a macrophage–adipocyte co-culture, the compound reduced the release of TNFα, IL-6, IL-1β, MCP-1 and RANTES, while enhancing glucose uptake and activation of AMPKα. Our results suggest that MTC produced anti-inflammatory and antioxidant activities in macrophages. MTC also prevented inflammation in macrophage–adipocyte co-culture. The effect of MTC on glucose uptake in adipocytes is proposed to be linked to activation of AMPK.

Moringa Fruit Inhibits LPS-Induced NO/iNOS Expression through Suppressing the NF-κB Activation in RAW264.7 Cells

2013 ◽  
Vol 41 (05) ◽  
pp. 1109-1123 ◽  
Author(s):  
Hyo-Jin Lee ◽  
Yun-Jeong Jeong ◽  
Tae-Sung Lee ◽  
Yoon-Yub Park ◽  
Whi-Gun Chae ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Inflammatory Mediators ◽  
Nuclear Translocation ◽  
Biologically Active ◽  
Raw264.7 Cells ◽  
No Production ◽  
Dependent Manner ◽  
Fruit Extract ◽  
Tnf Α ◽  
Anti Inflammatory

In this study, we evaluated the anti-inflammatory effects of moringa (Moringa oleifera Lam.), a natural biologically active substance, by determining its inhibitory effects on pro-inflammatory mediators in lipopolysaccharide (LPS)-stimulated macrophage RAW264.7 cells. Extracts from different parts of moringa (root, leaf, and fruit) reduced LPS-induced nitric oxide (NO) release in a dose-dependent manner. The moringa fruit extract most effectively inhibited LPS-induced NO production and levels of inducible nitric oxide synthase (iNOS). The moringa fruit extract also was shown to suppress the production of inflammatory cytokines including IL-1β, TNF-α, and IL-6. Furthermore, moringa fruit extract inhibited the cytoplasmic degradation of I κ B -α and the nuclear translocation of p65 proteins, resulting in lower levels of NF -κ B transactivation. Collectively, the results of this study demonstrate that moringa fruit extract reduces the levels of pro-inflammatory mediators including NO , IL-1β, TNF-α, and IL-6 via the inhibition of NF -κ B activation in RAW264.7 cells. These findings reveal, in part, the molecular basis underlying the anti-inflammatory properties of moringa fruit extract.

scholarly journals Gymnema inodorum (Lour.) Decne. Extract Alleviates Oxidative Stress and Inflammatory Mediators Produced by RAW264.7 Macrophages

2021 ◽  
Vol 2021 ◽  
pp. 1-20
Author(s):  
Benjawan Dunkhunthod ◽  
Chutima Talabnin ◽  
Mark Murphy ◽  
Kanjana Thumanu ◽  
Patcharawan Sittisart ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Nitric Oxide ◽  
Nuclear Translocation ◽  
Antioxidant Properties ◽  
Raw264.7 Cells ◽  
Mrna Levels ◽  
Ros Production ◽  
Raw264.7 Macrophages ◽  
Ifn Γ ◽  
Anti Inflammatory

Gymnema inodorum (Lour.) Decne. (G. inodorum) is widely used in Northern Thai cuisine as local vegetables and commercial herb tea products. In the present study, G. inodorum extract (GIE) was evaluated for its antioxidant and anti-inflammatory effects in LPS plus IFN-γ-induced RAW264.7 cells. Major compounds in GIE were evaluated using GC-MS and found 16 volatile compounds presenting in the extract. GIE exhibited antioxidant activity by scavenging the intracellular reactive oxygen species (ROS) production and increasing superoxide dismutase 2 (SOD2) mRNA expression in LPS plus IFN-γ-induced RAW264.7 cells. GIE showed anti-inflammatory activity through suppressing nitric oxide (NO), proinflammatory cytokine production interleukin 6 (IL-6) and also downregulation of the expression of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), and IL-6 mRNA levels in LPS plus IFN-γ-induced RAW264.7 cells. Mechanism studies showed that GIE suppressed the NF-κB p65 nuclear translocation and slightly decreased the phosphorylation of NF-κB p65 (p-NF-κB p65) protein. Our studies applied the synchrotron radiation-based FTIR microspectroscopy (SR-FTIR), supported by multivariate analysis, to identify the FTIR spectral changes based on macromolecule alterations occurring in RAW264.7 cells. SR-FTIR results demonstrated that the presence of LPS plus IFN-γ in RAW264.7 cells associated with the increase of amide I/amide II ratio (contributing to the alteration of secondary protein structure) and lipid content, whereas glycogen and other carbohydrate content were decreased. These findings lead us to believe that GIE may prevent oxidative damage by scavenging intracellular ROS production and activating the antioxidant gene, SOD2, expression. Therefore, it is possible that the antioxidant properties of GIE could modulate the inflammation process by regulating the ROS levels, which lead to the suppression of proinflammatory cytokines and genes. Therefore, GIE could be developed into a novel antioxidant and anti-inflammatory agent to treat and prevent diseases related to oxidative stress and inflammation.

Spiranthes sinensis Suppresses Production of Pro-Inflammatory Mediators by Down-Regulating the NF-κB Signaling Pathway and Up-Regulating HO-1/Nrf2 Anti-Oxidant Protein

2015 ◽  
Vol 43 (05) ◽  
pp. 969-989 ◽  
Author(s):  
Pei-Hsin Shie ◽  
Shyh-Shyun Huang ◽  
Jeng-Shyan Deng ◽  
Guan-Jhong Huang
Keyword(s):  
Inflammatory Mediators ◽  
Nuclear Translocation ◽  
Ethyl Acetate Fraction ◽  
Raw264.7 Cells ◽  
Prostaglandin E ◽  
Protein Levels ◽  
Tnf Α ◽  
Anti Oxidant ◽  
Spiranthes Sinensis

Spiranthes sinensis is an east Asian wild orchid used in Chinese folk medicine. In this study, an ethyl acetate fraction from S. sinensis(SSE) was found to suppress the production of LPS-stimulated inflammatory mediators in RAW264.7 cells and BALB/c mice. SSE inhibited the production of pro-inflammatory mediators such as nitric oxide (NO), prostaglandin E2 (PGE2), tumo necrosis factor-α (TNF-α), IL-1β, and IL-6 in LPS-stimulated RAW264.7 cells. SSE also significantly suppressed LPS-stimulated protein levels of iNOS and mPGES-1 by blocking IκB phosphorylation, NF-κB nuclear translocation, and MAPKs phosphorylation. In addition, SSE treatment also enhanced protein levels of HO-1 and anti-oxidant enzymes (SOD-1, CAT, and GPx-1) through the nuclear translocation of Nrf2 in LPS-stimulated RAW264.7 cells. In vivo, we demonstrated that SSE attenuated the levels of pro-inflammatory mediators (NO, TNF-α, IL-1β, and IL-6), ALT, and AST in the serum of LPS-stimulated BALB/c mice. Western blotting revealed that SSE enhanced HO-1 expression in lung and liver tissue after LPS injection in mice. These results suggest that the anti-inflammatory properties of SSE involve the suppression of iNOS, mPGES-1, and inflammatory mediators by inducing the HO-1 pathway in LPS-stimulated RAW264.7 cells and BALB/c mice.

scholarly journals Anti-inflammatory effects of DhHP-6 in LPS-induced RAW264.7 macrophages and carrageenan-induced air pouch model in rat

2021 ◽  
Author(s):  
Fanwei Meng ◽  
Junfeng Ke ◽  
Jia Xu ◽  
Jinze Li ◽  
Liping Wang
Keyword(s):  
Nuclear Translocation ◽  
Raw264.7 Cells ◽  
Total Proteins ◽  
Air Pouch ◽  
Inflammatory Agent ◽  
Raw264.7 Macrophages ◽  
Peptide Mimic ◽  
Anti Inflammatory ◽  
Air Pouch Model ◽  
Pro Inflammatory Cytokines

Abstract DhHP-6 (Deuterohemin-Ala-His-Thr-Val-Glu-Lys) is a novel peptide mimic of peroxidases that was previously designed in our laboratory. Here, we explored the anti-inflammatory potential of DhHP-6 against lipopolysaccharide(LPS)stimulated inflammatory response in RAW264.7 cells and carrageenan-induced air pouch model rats. DhHP-6 treatment dramatically attenuated the production of nitric oxide (NO), IL-6, andTNF-α in LPS induced RAW264.7 cells. Also, it blocked phosphorylation and degradation of IκBα and suppressed the nuclear translocation of p65. DhHP-6 (0.2, 0.6, and 2.0 mg/kg) significantly reduced the levels of total proteins and WBC counts in the exudates of the air pouch model rats. Moreover, MDA contents in the plasma of rats were reduced and SOD activities were enhanced in the DhHP-6-treatment group. Our results strongly show the effectiveness of DhHP-6 as an anti-inflammatory agent. The mechanism could be related to the reduction of Reactive oxygen species(ROS), inhibition of NF-κB nuclear translocation, and reduction of pro-inflammatory cytokines.

scholarly journals Benzoylaconine Modulates LPS-Induced Responses through Inhibition of Toll-Like Receptor-Mediated NF-κB and MAPK Signaling in RAW264.7 Cells

2021 ◽  
Author(s):  
Changkai Zhou ◽  
Jing Gao ◽  
Hongyan Ji ◽  
Wenjing Li ◽  
Xiaomin Xing ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Nuclear Translocation ◽  
Mapk Signaling ◽  
Dependent Manner ◽  
Toll Like Receptor ◽  
Protein Levels ◽  
Raw264.7 Macrophages ◽  
Elevated Protein ◽  
Anti Inflammatory ◽  
Inflammatory Effect

Abstract Previous studies have shown that benzoylaconine (BAC), a representative monoester alkaloid, has a potential anti-inflammatory effect. This study investigated the underlying molecular mechanisms using the mode of LPS-activated RAW264.7 macrophage cells. Our findings showed that BAC significantly suppressed the release of pro-inflammatory cytokines and mediators, including IL-6, TNF-α, IL-1β, ROS, NO, and PGE2. BAC treatment also effectively downregulated the elevated protein levels of iNOS and COX-2 induced by LPS in a dose-dependent manner. In this study, we found that BAC inhibited LPS-induced NF-κB activation by reducing the phosphorylation and degradation of IκBα by Western blotting and blocking the nuclear translocation of p65 using an immunofluorescence assay. The elevated protein levels of JNK, p38, and ERK phosphorylation after LPS stimulation were restored effectively by BAC treatment. Moreover, LPS-induced phosphorylation of TAK1, which is a crucial upstream regulatory factor of Toll-like receptor-induced MAPK and NF-κB signaling, was inhibited by BAC in activated RAW264.7 macrophages. These findings demonstrated that BAC exhibited an anti-inflammatory effect by inhibition of Toll-like receptor-induced MAPK and NF-κB pathways, indicating that it could potentially be used for treating inflammatory diseases.

scholarly journals Dehydrostephanine Isolated from Stephania venosa Possesses Anti-Inflammatory Activity in Lipopolysaccharide-Activated RAW264.7 Macrophages

2020 ◽  
Vol 17 (7) ◽  
pp. 655-664
Author(s):  
Wanatsanan CHULRIK ◽  
Chutima JANSAKUN ◽  
Waraluck CHAICHOMPOO ◽  
Janejira HATA ◽  
Poonsit HIRANSAI ◽  
...  
Keyword(s):  
Inflammatory Mediators ◽  
Raw264.7 Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Griess Reagent ◽  
Raw264.7 Macrophages ◽  
Ic50 Value ◽  
Anti Oxidant ◽  
Anti Inflammatory ◽  
Pre Treatment ◽  
Factor Α

Stephania venosa (Blume) Spreng. is a medicinal herb wildly used as a folklore medicine in Thailand. Many studies have reported that S. venosa tuber revealed a variety of pharmacological activities including anti-malarial, anti-microbial, anti-cancer, anti-oxidant, and anti-inflammatory activities. In this study, we investigated the effects of (–)-stephanine and dehydrostephanine isolated from S. venosa tuber on anti-inflammation in lipopolysaccharide (LPS)-activated RAW264.7 macrophages. RAW264.7 cells were treated with (–)-stephanine and dehydrostephanine in the presence of LPS and cell viability was determined by MTT assay. The levels of inflammatory mediators, nitric oxide (NO) and pro-inflammatory cytokines were determined by Griess reagent and enzyme-linked immunosorbent assay, respectively. Pre-treatment of dehydrostephanine significantly suppressed NO secretion in LPS-activated RAW264.7 cells with the half-maximal NO inhibitory concentration (IC50) value of 26.81±0.25 μM. However, (–)-stephanine had IC50 value on the inhibition of NO secretion of >40 μM. In addition, dehydrostephanine at concentrations of 20 - 80 μM significantly reduced LPS-induced tumor necrosis factor-α, interleukin-1b, and interleukin-6 production in RAW264.7 cells. The present study showed that dehydrostephanine possesses the anti-inflammatory effect on LPS-activated RAW264.7 macrophages by suppression of inflammatory mediators. Dehydrostephanine may be a promising candidate compound for further investigation of a novel class of anti-inflammatory drug.

scholarly journals Upregulation of miR-150-5p alleviates LPS-induced inflammatory response and apoptosis of RAW264.7 macrophages by targeting Notch1

2020 ◽  
Vol 15 (1) ◽  
pp. 544-552
Author(s):  
Xiaoyan Deng ◽  
Zhixing Lin ◽  
Chao Zuo ◽  
Yanjie Fu
Keyword(s):  
Inflammatory Response ◽  
Underlying Mechanism ◽  
Notch Receptor ◽  
Raw264.7 Cells ◽  
Luciferase Reporter ◽  
Raw264.7 Macrophages ◽  
Factor Α ◽  
Anti Inflammation ◽  
Necrosis Factor ◽  
Dual Luciferase Reporter Assay

AbstractCirculating miR-150-5p has been identified as a prognostic marker in patients with critical illness and sepsis. Herein, we aimed to further explore the role and underlying mechanism of miR-150-5p in sepsis. Quantitative real-time-PCR assay was performed to detect the expression of miR-150-5p upon stimulation with lipopolysaccharide (LPS) in RAW264.7 cells. The levels of tumor necrosis factor-α, interleukin (IL)-6 and IL-1β were measured by ELISA assay. Cell apoptosis was determined using flow cytometry. Western blot was used to assess notch receptor 1 (Notch1) expression in LPS-induced RAW264.7 cells. Dual-luciferase reporter assay was employed to validate the target of miR-150-5p. Our data showed that miR-150-5p was downregulated and Notch1 was upregulated in LPS-stimulated RAW264.7 cells. miR-150-5p overexpression or Notch1 silencing alleviated LPS-induced inflammatory response and apoptosis in RAW264.7 cells. Moreover, Notch1 was a direct target of miR-150-5p. Notch1 abated miR-150-5p-mediated anti-inflammation and anti-apoptosis in LPS-induced RAW264.7 cells. miR-150-5p alleviated LPS-induced inflammatory response and apoptosis at least partly by targeting Notch1 in RAW264.7 cells, highlighting miR-150-5p as a target in the development of anti-inflammation and anti-apoptosis drugs for sepsis treatment.

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