Fisetin inhibits lipopolysaccharide-induced inflammatory response by activating β-catenin, leading to a decrease in endotoxic shock

AbstractFisetin is a naturally occurring flavonoid that possesses several pharmacological benefits including anti-inflammatory activity. However, its precise anti-inflammatory mechanism is not clear. In the present study, we found that fisetin significantly inhibited the expression of proinflammatory mediators, such as nitric oxide (NO) and prostaglandin E2 (PGE2), and cytokines, such as interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α), in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Additionally, fisetin attenuated LPS-induced mortality and abnormalities in zebrafish larvae and normalized the heart rate. Fisetin decreased the recruitment of macrophages and neutrophils to the LPS-microinjected inflammatory site in zebrafish larvae, concomitant with a significant downregulation of proinflammatory genes, such as inducible NO synthase (iNOS), cyclooxygenase-2a (COX-2a), IL-6, and TNF-α. Fisetin inhibited the nuclear localization of nuclear factor-kappa B (NF-κB), which reduced the expression of pro-inflammatory genes. Further, fisetin inactivated glycogen synthase kinase 3β (GSK-3β) via phosphorylation at Ser9, and inhibited the degradation of β-catenin, which consequently promoted the localization of β-catenin into the nucleus. The pharmacological inhibition of β-catenin with FH535 reversed the fisetin-induced anti-inflammatory activity and restored NF-κB activity, which indicated that fisetin-mediated activation of β-catenin results in the inhibition of LPS-induced NF-κB activity. In LPS-microinjected zebrafish larvae, FH535 promoted the migration of macrophages to the yolk sac and decreased resident neutrophil counts in the posterior blood island and induced high expression of iNOS and COX-2a, which was accompanied by the inhibition of fisetin-induced anti-inflammatory activity. Altogether, the current study confirmed that the dietary flavonoid, fisetin, inhibited LPS-induced inflammation and endotoxic shock through crosstalk between GSK-3β/β-catenin and the NF-κB signaling pathways.

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Thiadiazolidinone-8 Ameliorates Inflammation Associated with Experimental Colitis in Mice

Pharmacology ◽

10.1159/000471808 ◽

2017 ◽

Vol 101 (1-2) ◽

pp. 35-42 ◽

Cited By ~ 2

Author(s):

Vanessa Mateus ◽

João Rocha ◽

Paula Alves ◽

Hélder Mota-Filipe ◽

Bruno Sepodes ◽

...

Keyword(s):

Glycogen Synthase ◽

Experimental Colitis ◽

Trinitrobenzene Sulfonic Acid ◽

Protective Actions ◽

Tnf Α ◽

Inflammatory Bowel ◽

Anti Inflammatory ◽

Gsk 3Β ◽

Necrosis Factor

Thiadiazolidinone-8 (TDZD-8) is an effective thiadiazolidinone derivate that is able to suppress the expression of inflammatory cytokines; it also presents tissue protective actions by glycogen synthase kinase (GSK)-3β inhibition, promoting thus an anti-inflammatory effect. Since inflammatory bowel disease is a chronic disease with reduced quality of life, where currently available therapies are only able to induce or maintain the patient in remission, it is crucial to investigate new pharmacological approaches. The main objective of this study was to evaluate the effect of TDZD-8 in 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis. Male CD-1 mice with TNBS-induced colitis were treated with a daily dose of TDZD-8 5 mg/kg/day IP during 4 days. The anti-inflammatory properties of TDZD-8 in the TNBS-induced colitis were confirmed by suppression of pro-inflammatory mediators, such as tumor necrosis factor (TNF)-α, interleukin (IL)-1β and myeloperoxidase, as well as by the significant increase of the anti-inflammatory cytokine, IL-10. These treated mice also presented a reduction in fecal hemoglobin and alkaline phosphatase, suggesting a beneficial effect of TDZD-8. Furthermore, renal and hepatic biomarkers remained stabilized after treatment. In conclusion, TDZD-8 reduces the inflammatory response associated with TNBS-induced colitis in mice, and modulation of GSK-3β seems to be an interesting pharmacological target in colitis.

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ANTI-INFLAMMATORY AND ANTI-GRANULOMA ACTIVITY OF SULFORAPHANE, A NATURALLY OCCURRING ISOTHIOCYANATE FROM BROCCOLI (BRASSICA OLERACEA)

Asian Journal of Pharmaceutical and Clinical Research ◽

10.22159/ajpcr.2018.v11i6.25279 ◽

2018 ◽

Vol 11 (6) ◽

pp. 411 ◽

Cited By ~ 1

Author(s):

Sangeeta Mohanty ◽

Abhisek Pal ◽

V Badireenath Konkimalla ◽

Sudam Chandra Si

Keyword(s):

Mononuclear Cells ◽

Inflammatory Activity ◽

Enzyme Linked Immunosorbent Assay ◽

Dependent Manner ◽

Serum Glutamic Oxaloacetic Transaminase ◽

Peripheral Blood Mononuclear ◽

Necrosis Factor Alpha ◽

Mediators Of Inflammation ◽

Tnf Α ◽

Anti Inflammatory

Objective: The objective of this study was to establish the anti-inflammatory activity of sulforaphane (SFN) in different acute and subchronic models of inflammation. Methods: The anti-inflammatory activity of SFN was evaluated by the secretion of proinflammatory cytokines in rat peripheral blood mononuclear cells (interleukin-6 [IL-6] and tumor necrosis factor alpha [TNF-α]) which are important mediators of inflammation as determined by enzyme-linked immunosorbent assay. Furthermore, paw volume was determined in various acute models of inflammation, and percentage inhibition of granuloma tissue was assessed by cotton pellet-induced granuloma model. From serum, serum glutamic pyruvic transaminase, serum glutamic oxaloacetic transaminase, and alkaline phosphatase levels were determined which is followed by assay for estimation of antioxidants such as superoxide dismutase (SOD), catalase, and glutathione (GSH). Results: SFN showed significant anti-inflammatory activity against paw edema induced by carrageenin/histamine/egg-albumin. A remarkable control in inflammation was observed most notably at the highest test dose of 5 mg/kg in the subchronic granuloma model. In addition, the release of inflammatory cytokines such as IL-6 and TNF-α which is responsible for inflammatory activity gets attenuated by SFN (∗p<0.05; ∗∗p< 0.01). Moreover, toxic control rats showed significant decreased levels of GSH, catalase, and SOD and increased the level of serum hepatic enzymes which gets reversed by SFN in dose-dependent manner. Conclusions: The present findings demonstrated that SFN can recover inflammation by inhibiting TNF-α and IL-6 in inflammation process.

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Antibacterial and Anti-Inflammatory Activities ofPhysalis Alkekengivar.franchetiiand Its Main Constituents

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2016/4359394 ◽

2016 ◽

Vol 2016 ◽

pp. 1-10 ◽

Cited By ~ 7

Author(s):

Zunpeng Shu ◽

Na Xing ◽

Qiuhong Wang ◽

Xinli Li ◽

Bingqing Xu ◽

...

Keyword(s):

Nitric Oxide ◽

Nuclear Translocation ◽

Interleukin 1 ◽

Inflammatory Activity ◽

Prostaglandin E ◽

Active Components ◽

Necrosis Factor Alpha ◽

Anti Inflammatory

This study was designed to determine whether the 50% EtOH fraction from AB-8 macroporous resin fractionation of a 70% EtOH extract ofP. Alkekengi(50-EFP) has antibacterial and/or anti-inflammatory activity bothin vivoandin vitroand to investigate the mechanism of 50-EFP anti-inflammatory activity. Additionally, this study sought to define the chemical composition of 50-EFP. Results indicated that 50-EFP showed significant antibacterial activityin vitroand efficacyin vivo. Moreover, 50-EFP significantly reduced nitric oxide (NO), prostaglandin E2(PGE2), tumor necrosis factor alpha (TNF-α), interleukin 1 (IL-1), and interleukin 6 (IL-6) production in lipopolysaccharide- (LPS-) stimulated THP-1 cells. Nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) (examined at the protein level) in THP-1 cells were suppressed by 50-EFP, which inhibited nuclear translocation of p65. Consistent with this anti-inflammatory activityin vitro, 50-EFP reduced inflammation in both animal models. Finally, seventeen compounds (8 physalins and 9 flavones) were isolated as major components of 50-EFP. Our data demonstrate that 50-EFP has antibacterial and anti-inflammatory activities bothin vitroandin vivo. The anti-inflammatory effect appears to occur, at least in part, through the inhibition of nuclear translocation of p65. Moreover, physalins and flavones are probably the active components in 50-EFP that exert antibacterial and anti-inflammatory activities.

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Staphylococcus aureus Induces Microglial Inflammation via a Glycogen Synthase Kinase 3β-Regulated Pathway

Infection and Immunity ◽

10.1128/iai.00176-09 ◽

2009 ◽

Vol 77 (9) ◽

pp. 4002-4008 ◽

Cited By ~ 31

Author(s):

Yi-Lin Cheng ◽

Chi-Yun Wang ◽

Wei-Ching Huang ◽

Cheng-Chieh Tsai ◽

Chia-Ling Chen ◽

...

Keyword(s):

Staphylococcus Aureus ◽

Nuclear Translocation ◽

Glycogen Synthase ◽

Glycogen Synthase Kinase ◽

No Production ◽

Glycogen Synthase Kinase 3Β ◽

Necrosis Factor Alpha ◽

Tnf Α ◽

Gsk 3Β ◽

Microglial Inflammation

ABSTRACT A proinflammatory role for glycogen synthase kinase 3β (GSK-3β) has been demonstrated. Here, we addressed its roles on heat-inactivated Staphylococcus aureus-induced microglial inflammation. Heat-inactivated S. aureus induced tumor necrosis factor alpha (TNF-α) and nitric oxide (NO) production, at least in part, via a Toll-like receptor 2-regulated pathway. Neutralization of TNF-α largely blocked heat-inactivated S. aureus-induced NO. Heat-inactivated S. aureus activated GSK-3β, and inhibiting GSK-3β reduced TNF-α production as well as inducible NO synthase (iNOS)/NO biosynthesis. While activation of NF-κB was essential for heat-inactivated S. aureus-induced TNF-α and NO, inhibiting GSK-3β blocked heat-inactivated S. aureus-induced NF-κB p65 nuclear translocation. Additionally, inhibiting GSK-3β enhanced heat-inactivated S. aureus-induced interleukin-10 (IL-10) production (IL-10 is an anti-inflammatory cytokine which inhibits TNF-α production). Neutralization of IL-10 reduced TNF-α downregulation caused by GSK-3β inhibition. These results suggest that GSK-3β regulates heat-inactivated S. aureus-induced TNF-α and NO production in microglia mainly by activating NF-κB and probably by inhibiting IL-10.

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Diverse Krill Lipid Fractions Differentially Reduce LPS-Induced Inflammatory Markers in RAW264.7 Macrophages In Vitro

Foods ◽

10.3390/foods10112887 ◽

2021 ◽

Vol 10 (11) ◽

pp. 2887

Author(s):

Dan Xie ◽

Fangyuan He ◽

Xiaosan Wang ◽

Xingguo Wang ◽

Qingzhe Jin ◽

...

Keyword(s):

Nitric Oxide ◽

Inflammatory Activity ◽

Chemical Compositions ◽

Krill Oil ◽

Proinflammatory Mediators ◽

Raw264.7 Macrophages ◽

Epa And Dha ◽

Tnf Α ◽

Lipid Fractions ◽

Anti Inflammatory

Antarctic krill oil is an emerging marine lipid and expected to be a potential functional food due to its diverse nutrients, such as eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), phospholipids, astaxanthin and tocopherols. Although krill oil has been previously proved to have anti-inflammatory activity, there is little information about the relationship between its chemical compositions and anti-inflammatory activity. In this study, the RAW264.7 macrophages model was used to elucidate and compare the anti-inflammatory potential of different krill lipid fractions: KLF-A, KLF-H and KLF-E, which have increasing phospholipids, EPA and DHA contents but decreasing astaxanthin and tocopherols levels. Results showed that all the krill lipid fractions alleviated the inflammatory reaction by inhibition of production of nitric oxide (NO), release of tumor necrosis factor-α (TNF-α), interleukin (IL)-1β and IL-6 and gene expression of proinflammatory mediators including TNF-α, IL-1β, IL-6, cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS). In addition, KLF-E with the highest phospholipids, EPA and DHA contents showed the strongest inhibition effect on the LPS-induced proinflammatory mediator release and their gene expressions. The results would be helpful to provide powerful insights into the underlying anti-inflammatory mechanism of krill lipid and guiding the production of krill oil products with tailor-made anti-inflammatory activity.

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Aktivitas Antiinflamasi Crude Extract Fukoidan dari Sargassum crassifolium pada Sel RAW 264.7 yang Diinduksi LPS

Jurnal Kefarmasian Indonesia ◽

10.22435/jki.v9i2.1547 ◽

2019 ◽

pp. 97-105

Author(s):

Pangartika Hikariastri ◽

Hendig Winarno ◽

Kusmardi Kusmardi ◽

Dian Ratih Laksmitawati ◽

Syamsudin Abdillah

Keyword(s):

Crude Extract ◽

Macrophage Activation ◽

Basic Mechanism ◽

Inflammatory Activity ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Proinflammatory Mediators ◽

Promising Target ◽

Tnf Α ◽

Anti Inflammatory

Inflammation is known as the basic mechanism underlying various chronic diseases. Macrophage activation by inflammatory stimulus induces the release of inflammatory mediators, thus the mediators becoming a promising target of anti-inflammatory drug development. Previous studies indicated that fucoidan has anti-inflammatory activity by inhibiting the release of proinflammatory mediators. The aim of this study is determining anti-inflammatory activity of fucoidan crude extract form Sargassum crassifolium Garut waters by observing its effect on proinflammatory cytokines TNF-α, IL-1β, dan IL-6. Fucoidan is a polysaccharide substance which has various characteristics, depending on the source and the extraction method which is influencing its bioactivity. Sargassum crassifolium collected from Garut-West Java is extracted using diluted HCl 0,1 M and precipitated with ethanol to obtain fucoidan crude extract. The crude extract is tested on LPS-induced RAW 264.7 cells to evaluate its effect on TNF-α, IL-1β, and IL-6 level using ELISA method. The result showed that fucoidan crude extract decreased the level of TNF-α by the dose of 25 and 50 μg/ml, and decreased the level of IL-1β and IL-6 by the dose of 25 μg/ml. The dose of 50 μg/ml failed to inhibit IL-1β and IL-6 production. This study showed that fucoidan crude extract derived from S. crassifolium has anti-inflammatory activity to RAW 264.7 cells by the dose of 25 μg/ml.

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Anti-inflammatory Effect of Woodfordia fructicosa Leaves Ethanolic Extract on Adjuvant and Carragenan Treated Rats

Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry ◽

10.2174/1871523018666190222120127 ◽

2020 ◽

Vol 19 (2) ◽

pp. 103-112

Author(s):

Hem Raj ◽

Avneet Gupta ◽

Neeraj Upmanyu

Keyword(s):

Animal Models ◽

Ethanolic Extract ◽

Inhibitory Effect ◽

Inflammatory Activity ◽

Paw Edema ◽

Sprague Dawley ◽

Traditional Use ◽

Necrosis Factor Alpha ◽

Tnf Α ◽

Anti Inflammatory

Background: Woodfordia fructicosa is used traditionally for the treatment of inflammation associated with arthritis. Methods: In the present study, the anti-inflammatory activity of W. fructicosa (WFE) leaves ethanolic extract was assessed in Sprague Dawley rats by giving 200 mg/kg dose orally. Inflammation was studied by using carrageenan induced paw edema, Freund’s adjuvant (FA) and monosodium iodo acetate (MIA) induced arthritis as animal models. Serum tumor necrosis factor-alpha (TNF-α) was estimated in blood sample of animals treated with FA. The one way ANOVA followed by Bonferroni’s test was used for statistical analysis. Results: WFE significantly decreased (P<0.05, P<0.001) paw thickness in carrageenan induced paw edema and FA induced arthritis. The significant decrease in knee diameter (P<0.001) in MIA induced arthritis as well as inhibitory effect (P<0.001) on elevated TNF- α was observed. Conclusion: These results showed that the WFEexerted an inhibitory effect on TNF-α and carrageenan paw edema which may justify its traditional use in inflammatory conditions. Thus, the study shows that leaves of W. fruticose afford anti-inflammatory activity by preventing the inflammation in different animal models.

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In vitro evaluation of selligueain A effects on the proinﬂammatory mediators production in RAW264.7 murine macrophages

Journal of Herbmed Pharmacology ◽

10.34172/jhp.2021.36 ◽

2021 ◽

Vol 10 (3) ◽

pp. 313-318

Author(s):

Deden Winda Suwandi ◽

Tina Rostnawati ◽

Muchtaridi Muchtaridi ◽

Anas Subarnas

Keyword(s):

Nitric Oxide ◽

Inflammatory Mediators ◽

Cell Model ◽

Inflammatory Activity ◽

Raw264.7 Cells ◽

Proinflammatory Mediators ◽

Tnf Α ◽

Anti Inflammatory

Introduction: Selligueain A derived from the roots of Polypodium feei was shown to have anti-inflammatory activity, which was tested in vivo on the rats’ paw edema induced by carrageenan. The aim of this study was to evaluate the anti-inflammatory mechanism of selligueain A in vitro against the production of pro-inflammatory mediators. Methods: In this study, RAW264.7 cells were used as an inflammatory cell model, and observations were made on the inflammatory mediators nitric oxide (NO), inducible nitric oxide synthase (iNOS), and tumour necrosis factor-α (TNF-α). The NO concentration was measured by the Griess reaction, and the iNOS enzyme and the TNF-α concentrations were determined by the ELISA method. Cell viability was assessed by the [3-(4,5-dimethylthiazol-2yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium] (MTS) test. Results: Selligueain A at concentrations of 100 and 150 µM suppressed the production of NO, iNOS, and TNF-α in RAW264.7 cells stimulated by lipopolysaccharide (LPS). The concentration of 150 µM showed the highest inhibition of NO, iNOS, and TNF-α mediators with the percentage inhibition of 64.85, 55.01, and 48.54%, respectively. Conclusion: This study shows that selligueain A has anti-inflammatory activity through inhibition of NO, iNOS, and TNF-α production in RAW264.7 macrophage cells.

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Anti-Neuroinflammatory and Anti-Inflammatory Activities of Phenylheptatriyne Isolated from the Flowers of Coreopsis lanceolata L. via NF-κB Inhibition and HO-1 Expression in BV2 and RAW264.7 Cells

International Journal of Molecular Sciences ◽

10.3390/ijms22147482 ◽

2021 ◽

Vol 22 (14) ◽

pp. 7482

Author(s):

Hwan Lee ◽

Zhiming Liu ◽

Chi-Su Yoon ◽

Linsha Dong ◽

Wonmin Ko ◽

...

Keyword(s):

Silica Gel ◽

Column Chromatography ◽

Raw264.7 Cells ◽

Necrosis Factor Alpha ◽

Tnf Α ◽

Etoac Fraction ◽

Factor Alpha ◽

Anti Inflammatory ◽

And Oxidative Stress ◽

Necrosis Factor

Aging is associated with immune disregulation and oxidative stress which lead to inflammation and neurodegenerative diseases. We have tried to identify the anti-neuroinflammatory and anti-inflammatory components of Coreopsis lanceolata L. The dried flowers of C. lanceolata were extracted with 70% EtOH, and the obtained extract was divided into CH2Cl2, EtOAc, n-BuOH, and H2O fractions. The CH2Cl2 fraction was separated using silica gel and C-18 column chromatography to yield phenylheptatriyne (1), 2′-hydroxy-3,4,4′-trimethoxychalcone (2), and 4′,7-dimethoxyflavanone (3). Additionally, the EtOAc fraction was subjected to silica gel, C-18, and Sephadex LH-20 column chromatography to yield 8-methoxybutin (4) and leptosidin (5). All the compounds isolated from C. lanceolata inhibited the production of nitric oxide (NO) in LPS-induced BV2 and RAW264.7 cells. In addition, phenylheptatriyne and 4′,7-dimethoxyflavanone reduced the secretion of inflammatory cytokines, tumor necrosis factor alpha (TNF-α), and interleukin (IL)-6. Among them, phenylheptatriyne was significantly downregulated in the expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2). Subsequently, phenylheptatriyne also effectively inhibited nuclear factor-kappa B (NF-κB) activation in LPS-stimulated BV2 and RAW264.7 cells. Based on these results, the anti-neuroinflammatory effect of phenylheptatriyne isolated from C. lanceolata was confirmed, which may exert a therapeutic effect in treatment of neuroinflammation-related diseases.

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Processed Scutellaria baicalensis Georgi Extract Alleviates LPS-Induced Inflammatory and Oxidative Stress through a Crosstalk between NF-κB and KEAP1/NRF2 Signaling in Macrophage Cells

Applied Sciences ◽

10.3390/app11136055 ◽

2021 ◽

Vol 11 (13) ◽

pp. 6055

Author(s):

Akhtar Ali ◽

En-Hyung Kim ◽

Jong-Hyun Lee ◽

Kang-Hyun Leem ◽

Shin Seong ◽

...

Keyword(s):

Scutellaria Baicalensis ◽

Raw 264.7 Cells ◽

Prostaglandin E ◽

Scutellaria Baicalensis Georgi ◽

Anticancer Effects ◽

Tnf Α ◽

Clinical Benefits ◽

Anti Oxidant ◽

Cox 2 ◽

Anti Inflammatory

Prolonged inflammation results in chronic diseases that can be associated with a range of factors. Medicinal plants and herbs provide synergistic benefits based on the interaction of multiple phytochemicals. The dried root of Scutellaria baicalensis Georgi and its compounds possess anti-inflammatory, anti-oxidative, and anticancer effects. Processing is a traditional method to achieve clinical benefits by improving therapeutic efficacy and lowering toxicity. In this study, we investigated the anti-inflammatory and anti-oxidant effect of processed Scutellaria baicalensis Georgi extract (PSGE) against lipopolysaccharide (LPS) stimulated RAW 264.7 cells. Data using Griess assay and ELISA showed that PSGE decreased nitric oxide and prostaglandin E2 (PGE2) levels against LPS. PSGE treatment up-regulated 15-hydroxyprostaglandin dehydrogenase (PGDH), while cyclooxygenase (COX)-2 and microsomal prostaglandin E synthase (mPGES)-1 expression did not change. Interestingly, PGE2 inhibition was regulated by prostaglandin catabolic enzyme 15-PGDH rather than COX-2/mPGES-1, enzymes essential for PGE2 synthesis. Additionally, PSGE-suppressed LPS-induced IL-6 and TNF-α production through NF-κB signaling. NF-κB release from an inactive complex was inhibited by HO-1 which blocked IκBα phosphorylation. The ROS levels lowered by PSGE were measured with the H2DCFDA probe. PSGE activated NRF2 signaling and increased antioxidant Hmox1, Nqo1, and Txn1 gene expression, while reducing KEAP1 expression. In addition, pharmacological inhibition of HO-1 confirmed that the antioxidant enzyme induction by PSGE was responsible for ROS reduction. In conclusion, PSGE demonstrated anti-inflammatory and anti-oxidant effects due to NRF2/HO-1-mediated NF-κB and ROS inhibition.

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