scholarly journals Tocotrienol-rich Fraction Modulated Genes Responsible for Inflammation in Lipopolysaccharide-stimulated RAW 264.7 Macrophages

2021 ◽  
pp. 106-118
Author(s):  
Sitti Rahma Abd Hafid ◽  
Maliya Azilah Mohammad Aini ◽  
Nabiha Iran ◽  
Irmaliayana Norisam ◽  
Khairul Adzfa Radzun ◽  
...  
Keyword(s):  
Fold Change ◽  
Alpha Tocopherol ◽  
In Vivo Studies ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Necrosis Factor Alpha ◽  
Raw 264.7 Macrophages ◽  
Raw264.7 Macrophages ◽  
Tocotrienol Rich Fraction

Background: Inflammation plays a vital role in the pathogenesis of chronic non-communicable diseases (NCDs), the leading health issue worldwide. An earlier study reported that tocotrienol-rich fraction (TRF) showed better anti-inflammation effects in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. Aim: This study aimed to investigate the anti-inflammatory effects of tocotrienol-rich fraction at the molecular level by looking at the genes that were differentially regulated and pathways affected in LPS-stimulated macrophages exposed to TRF using the microarray approach. Methods: A microarray study was carried out in LPS-stimulated RAW 264.7 macrophages. Total ribonucleic acid (RNA) was extracted from the RAW 264.7 cells treated with TRF (10µg/mL), alpha-tocopherol (10 µg/mL) or LPS (10 ng/mL). Untreated cells served as control. Enrichment analyses, such as Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes pathway (KEGG), were conducted for genes listed in the differentially expressed genes (DEGs).  Results: The microarray analysis showed that the expression of five genes [Hamp, Interleukin-1a (IL-1a), IL-b, C-X-C motif chemokine ligand 2 (CXCL2) and colony-stimulating factor 3 (CSF3)] and one gene (SLC1A4), an amino acid transporter, was modulated (fold change 2, P< 0.05) in the TRF-treated cells. With a more stringent analysis (fold change 3, P < 0.05), only one gene (CSF3) was downregulated in the TRF-treated in RAW 264.7 cells. Analysis using the GO and KEGG pathways revealed interactions between pro-inflammatory agents such as tumor necrosis factor-alpha (TNFα) and nuclear factor kappa-light-chain-enhancer of activated B-cells (NF-B), as well as signaling pathways of interleukin (IL)-1 and IL-17. Conclusion: TRF modulated the expression of genes responsible for acute and chronic inflammation that were part of the lipoxygenase (LOX) and cyclooxygenase (COX) inflammatory pathways. Further investigation on the effects of TRF in different cell lines and in vivo studies should be conducted in the future.

scholarly journals MicroRNA-181a-5p regulates inflammatory response of macrophages in sepsis

Open Medicine ◽  
2019 ◽  
Vol 14 (1) ◽  
pp. 899-908 ◽  
Author(s):  
Zheng Huang ◽  
Hang Xu
Keyword(s):  
Inflammatory Response ◽  
Inflammatory Cytokines ◽  
Sirtuin 1 ◽  
Raw 264.7 Cells ◽  
Inflammatory Factors ◽  
Raw 264.7 ◽  
Raw264.7 Macrophages ◽  
Pathway Activation

AbstractThe aim of this study was to evaluate the role of miR-181a-5p in sepsis, and to further explore the molecular mechanism. RAW 264.7 cells were stimulated with 1 μg/ml LPS for 4 hours. Firstly, qRT-PCR and ELISA was adopted to evaluate the expression of miR-181a-5p and p ro-inflammatory cytokines in RAW 264.7 macrophages a fter LPS stimulation. Results showed that pro-inflammatory cytokines and miR-181a-5p were significantly increased after LPS treatment. Then, we identified that sirtuin-1 (SIRT1) was a direct target of miR-181a-5p and it was down-regulated in LPS treated RAW264.7 macrophages. Furthermore, the data suggested that the miR-181a-5p inhibitor significantly inhibited LPS enhanced inflammatory cytokines expression and NF-κB pathway activation, and these changes were eliminated by SIRT1 silencing. Moreover, the role of the miR-181a-5p inhibitor on sepsis was studied in vivo. We found that the miR-181a-5p inhibitor significantly decreased the secretion of inflammatory factors, and the levels of creatine (Cr), blood urea nitrogen (BUN), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in a serum for mice with sepsis. However, all the effects were reversed by SIRT1-siRNA. In summary, these results indicated that miR-181a-5p was involved in sepsis through regulating the inflammatory response by targeting SIRT1, suggesting that miR-181a-5p may be a potential target for the treatment of sepsis.

Carpesium macrocephalum Attenuates Lipopolysaccharide-Induced Inflammation in Macrophages by Regulating the NF-κB/IκB-α, Akt, and STAT Signaling Pathways

2013 ◽  
Vol 41 (04) ◽  
pp. 927-943 ◽  
Author(s):  
Sushruta Koppula ◽  
Wan-Jae Kim ◽  
Jun Jiang ◽  
Do-Wan Shim ◽  
Na-Hyun Oh ◽  
...  
Keyword(s):  
Inflammatory Responses ◽  
In Vivo Studies ◽  
Raw264.7 Cells ◽  
Traditional Use ◽  
Necrosis Factor Alpha ◽  
Proinflammatory Mediators ◽  
Raw264.7 Macrophages ◽  
Anti Inflammatory

Carpesium macrocephalum (CM) Fr. et Sav. (Compositae) has been used in Chinese folk medicine as an analgesic, hemostatic, antipyretic, and to suppress inflammatory conditions. In the present study we aimed to provide scientific evidence for the anti-inflammatory properties of CM extract and evaluate the intrinsic mechanisms involved in both in vitro and in vivo experimental models. In in vitro findings, CM significantly inhibited the LPS-stimulated release of proinflammatory mediators such as nitric oxide, tumor necrosis factor-alpha, prostaglandin E2, and interleukin-6 in RAW264.7 macrophages in a concentration-dependent fashion. The attenuation of inflammatory responses in LPS-activated RAW264.7 cells by CM was closely associated with the suppression of nuclear factor-kappa B (NF-κB) phosphorylation, IκB-α degradation, and phosphorylation of Akt. CM treatment also attenuated the phosphorylation of STAT through TRIF dependent pathways in LPS-activated RAW264.7 cells. In vivo studies revealed that CM extract concentration dependently suppressed the acetic acid-induced vascular permeability in mice. Considering the data obtained regulation of multiple signaling mechanisms involving TRIF and Akt/NF-κB pathways might be responsible for the potent anti-inflammatory action of CM, substantiating its traditional use in inflammatory diseases.

scholarly journals Anti-Inflammatory Comparison of Melatonin and Its Bromobenzoylamide Derivatives in Lipopolysaccharide (LPS)-Induced RAW 264.7 Cells and Croton Oil-Induced Mice Ear Edema

Molecules ◽  
2021 ◽  
Vol 26 (14) ◽  
pp. 4285
Author(s):  
Pimpichaya Sangchart ◽  
Panyada Panyatip ◽  
Teerasak Damrongrungruang ◽  
Aroonsri Priprem ◽  
Pramote Mahakunakorn ◽  
...  
Keyword(s):  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Dependent Manner ◽  
In Vivo Models ◽  
Raw 264.7 Macrophages ◽  
Ear Edema ◽  
Croton Oil ◽  
Anti Inflammatory

The pineal gland is a neuroendocrine organ that plays an important role in anti-inflammation through the hormone melatonin. The anti-inflammatory effects of melatonin and its derivatives have been reported in both in vitro and in vivo models. Our previous study reported the potent antioxidant and neuroprotective activities of bromobenzoylamide substituted melatonin. In silico analysis successfully predicted that melatonin bromobenzoylamid derivatives were protected from metabolism by CYP2A1, which is a key enzyme of the melatonin metabolism process. Therefore, the anti-inflammatory activities of melatonin and its bromobenzoylamide derivatives BBM and EBM were investigated in LPS-induced RAW 264.7 macrophages and croton oil-induced ear edema in mice. The experiments showed that BBM and EBM significantly reduced production of the inflammatory mediators interleukin-6 (IL-6), prostaglandin E2 (PGE2), and nitric oxide (NO) in a dose-dependent manner, but only slightly affected TNF-α in LPS-induced RAW 264.7 macrophages. This suggests that modifying melatonin at either the N1-position or the N-acetyl side chain affected production of NO, PGE2 and IL-6 in in vitro model. In the croton oil-induced mouse ear edema model, BBM, significantly decreased ear edema thickness at 2–4 h. It leads to conclude that bromobenzoylamide derivatives of melatonin may be one of the potential candidates for a new type of anti-inflammatory agent.

Anti-Inflammatory Effect of Local Anaesthetic Ropivacaine in Lipopolysaccharide-Stimulated RAW264.7 Macrophages

Pharmacology ◽  
2019 ◽  
Vol 103 (5-6) ◽  
pp. 228-235 ◽  
Author(s):  
Li Wu ◽  
Lu Li ◽  
Fang Wang ◽  
Xianwei Wu ◽  
Xin Zhao ◽  
...  
Keyword(s):  
Significant Inhibition ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Raw 264.7 Macrophages ◽  
Raw264.7 Macrophages ◽  
Tnf Α ◽  
Mitogen Activated Protein ◽  
Anti Inflammatory ◽  
Necrosis Factor ◽  
Inflammatory Effect

The present manuscript intended to investigate the anti-inflammatory effect of ropivacaine on lipopolysaccharide-induced inflammation in RAW 264.7 macrophages. Results suggested that ropivacaine causes significant inhibition of generation of nitric oxide (NO), prostaglandin E2, tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β, as well as expression of their synthesizing enzymes, inducible NO synthase, and cyclooxygenase-2. Moreover, ropivacaine causes inhibition of mitogen-activated protein kinases as well as nuclear factor-kappa B signaling pathway and apoptosis in RAW 264.7 cells. Based on the results, it has been suggested that ropivacaine showed beneficial effect against inflammation.

scholarly journals Aspergillus fumigatus Fumagillin Contributes to Host Cell Damage

2021 ◽  
Vol 7 (11) ◽  
pp. 936
Author(s):  
Xabier Guruceaga ◽  
Uxue Perez-Cuesta ◽  
Aize Pellon ◽  
Saioa Cendon-Sanchez ◽  
Eduardo Pelegri-Martinez ◽  
...  
Keyword(s):  
Aspergillus Fumigatus ◽  
Cell Damage ◽  
A549 Cells ◽  
In Vivo Studies ◽  
Raw 264.7 ◽  
Raw 264.7 Macrophages ◽  
Fungal Strains ◽  
Proliferation Ability

The activity of fumagillin, a mycotoxin produced by Aspergillus fumigatus, has not been studied in depth. In this study, we used a commercial fumagillin on cultures of two cell types (A549 pneumocytes and RAW 264.7 macrophages). This toxin joins its target, MetAP2 protein, inside cells and, as a result, significantly reduces the electron chain activity, the migration, and the proliferation ability on the A549 cells, or affects the viability and proliferation ability of the RAW 264.7 macrophages. However, the toxin stimulates the germination and double branch hypha production of fungal cultures, pointing out an intrinsic resistant mechanism to fumagillin of fungal strains. In this study, we also used a fumagillin non-producer A. fumigatus strain (∆fmaA) as well as its complemented strain (∆fmaA::fmaA) and we tested the fumagillin secretion of the fungal strains using an Ultra High-Performance Liquid Chromatography (UHPLC) method. Furthermore, fumagillin seems to protect the fungus against phagocytosis in vitro, and during in vivo studies using infection of immunosuppressed mice, a lower fungal burden in the lungs of mice infected with the ∆fmaA mutant was demonstrated.

scholarly journals Suppression of inflammation by ethanol extract of Clausena lansium via modulation of TLR4/MYD88/TRAF6 signaling pathway in RAW 264.7 macrophages

2019 ◽  
Vol 17 ◽  
pp. 205873921984197
Author(s):  
Guihong Huang ◽  
Juan Li ◽  
Wanlian Li ◽  
Tianxu Liu ◽  
Guojun Jiang ◽  
...  
Keyword(s):  
Ethanol Extract ◽  
Raw 264.7 Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Raw 264.7 ◽  
Rt Pcr ◽  
Anticancer Activities ◽  
Necrosis Factor Alpha ◽  
Raw 264.7 Macrophages ◽  
Protein Levels ◽  
Tnf Α

The fruit and root of Clausena lansium are remedy for bronchitis in oriental traditional medicine. Extracts from Clausena lansium are reported to have antioxidant, anti-inflammatory, and anticancer activities. This study is designed to investigate whether the ethanol extract of Clausena lansium (Lour.) Skeels could inhibit the lipopolysaccharides (LPS)-induced inflammatory in RAW 264.7 macrophages and the underlining mechanisms. Enzyme-linked immunosorbent assay (ELISA) and reverse transcription polymerase chain reaction (RT-PCR) are used to detect the secretion of tumor necrosis factor alpha (TNF-α) protein and expression of TNF-α mRNA, respectively. RT-PCR and Western blotting are used to determine the mRNA and protein levels of TLR4, MYD88, and TRAF6, respectively. The extract of Clausena lansium suppressed the expression and secretion of TNF-α, which is released by RAW 264.7 cells after LPS induction. Meanwhile, the expression of TLR4, MYD88, and TRAF6 are decreased by extracts from Clausena lansium. Meanwhile, NBP2-29328, an inhibitor of MYD88, synergistically enhances the inhibiting effect of extract from Clausena lansium. The results imply that ethanol extract from Clausena lansium attenuate macrophage-mediated inflammation and TLR4/MYD88/TRAF6 pathway is its potential target.

scholarly journals Comparison of CRISPR and adenovirus-mediated Myd88 knockdown in RAW 264.7 cells and responses to lipopolysaccharide stimulation

2021 ◽  
Vol 8 (3) ◽  
pp. e151
Author(s):  
Alexander L. Kolb ◽  
Marinaliz Reynoso ◽  
Ronald W. Matheny
Keyword(s):  
Genetic Manipulation ◽  
Interleukin 10 ◽  
Primary Response ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Necrosis Factor Alpha ◽  
Raw 264.7 Macrophages ◽  
Inflammatory Stimuli ◽  
Adenoviral Transduction ◽  
Nfκb Pathway

Genomic manipulation offers the possibility for novel therapies in lieu of medical interventions in use today. The ability togenetically restore missing inflammatory genes will have a monumental impact on our current immunotherapy treatments. This study compared the efficacy of two different genetic manipulation techniques: clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9) transfection to adenoviral transduction to determine which method would provide the most transient and stable knockdown of myeloid differentiation primary response 88 (MyD88). MyD88 is a major regulator of nuclear factor kappa light chain enhancer of activated B cells (NFκB) pathway in Raw 264.7 macrophages. Following genetic manipulation, cells were treated for 24 h with Lipopolysaccharide (LPS) to stimulate the inflammatory pathway. Confirmation of knockdown was determined by western immunoblotting and quantification of band density. Both CRISPR/Cas9 and adenoviral transduction produced similar knockdown efficiency (~64% and 60%, respectively) in MyD88 protein 48 h post adenoviral transduction. NFκB phosphorylation was increased in CRISPR/Cas9-mediated MyD88 knockdown and control cells, but not in adenovirus-mediated MyD88 knockdown cells, following LPS administration. CRISPR/Cas9-mediated MyD88 knockdown macrophages treated with LPS for 24 h showed a 65% reduction in tumor necrosis factor alpha (TNFα) secretion, and a 67% reduction in interleukin-10 (IL-10) secretion when compared to LPS-stimulated control cells (P ≤ 0.01 for both). LPS did not stimulate TNFα or IL-10 secretion in adenovirus-mediated control or MyD88 knockdown cells. These data demonstrate that Raw 264.7 macrophages maintain responsiveness to inflammatory stimuli following CRISPR/Cas9-mediated reductions in MyD88, but not following adenovirus-mediated MyD88 knockdown.

scholarly journals In Vitro and In Vivo Anti-Inflammatory Effects of Sulfated Polysaccharides Isolated from the Edible Brown Seaweed, Sargassum fulvellum

Marine Drugs ◽  
2021 ◽  
Vol 19 (5) ◽  
pp. 277
Author(s):  
Lei Wang ◽  
Hye-Won Yang ◽  
Ginnae Ahn ◽  
Xiaoting Fu ◽  
Jiachao Xu ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Brown Seaweed ◽  
Sulfated Polysaccharides ◽  
Raw 264.7 ◽  
Raw 264.7 Macrophages ◽  
Sargassum Fulvellum ◽  
Pharmaceutical Industries ◽  
Anti Inflammatory

In the present study, the in vitro and in vivo anti-inflammatory effects of the sulfated polysaccharides isolated from Sargassum fulvellum (SFPS) were evaluated in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages and zebrafish. The results indicated that SFPS improved the viability of LPS-stimulated RAW 264.7 macrophages from 80.02 to 86.80, 90.09, and 94.62% at the concentration of 25, 50, and 100 µg/mL, respectively. Also, SFPS remarkably and concentration-dependently decreased the production levels of inflammatory molecules including nitric oxide (NO), tumor necrosis factor-alpha, prostaglandin E2, interleukin-1 beta, and interleukin-6 in LPS-treated RAW 264.7 macrophages. In addition, SFPS significantly inhibited the expression levels of cyclooxygenase-2 and inducible nitric oxide synthase in LPS-treated RAW 264.7 macrophages. Furthermore, the in vivo test results indicated that SFPS improved the survival rate of LPS-treated zebrafish from 53.33 to 56.67, 60.00, and 70.00% at the concentration of 25, 50, and 100 µg/mL, respectively. In addition, SFPS effectively reduced cell death, reactive oxygen species, and NO levels in LPS-stimulated zebrafish. Taken together, these results suggested that SFPS possesses strong in vitro and in vivo anti-inflammatory activities, and could be used as an ingredient to develop anti-inflammatory agents in the functional food and pharmaceutical industries.

scholarly journals Phaseolin Attenuates Lipopolysaccharide-Induced Inflammation in RAW 264.7 Cells and Zebrafish

Biomedicines ◽  
2021 ◽  
Vol 9 (4) ◽  
pp. 420
Author(s):  
Su-Jung Hwang ◽  
Ye-Seul Song ◽  
Hyo-Jong Lee
Keyword(s):  
Nitric Oxide ◽  
Nuclear Translocation ◽  
Raw 264.7 Cells ◽  
Network Pharmacology ◽  
Raw 264.7 ◽  
Dependent Manner ◽  
Bioactive Constituents

Kushen (Radix Sophorae flavescentis) is used to treat ulcerative colitis, tumors, and pruritus. Recently, phaseolin, formononetin, matrine, luteolin, and quercetin, through a network pharmacology approach, were tentatively identified as five bioactive constituents responsible for the anti-inflammatory effects of S. flavescentis. However, the role of phaseolin (one of the primary components of S. flavescentis) in the direct regulation of inflammation and inflammatory processes is not well known. In this study, the beneficial role of phaseolin against inflammation was explored in lipopolysaccharide (LPS)-induced inflammation models of RAW 264.7 macrophages and zebrafish larvae. Phaseolin inhibited LPS-mediated production of nitric oxide (NO) and the expression of inducible nitric oxide synthase (iNOS), without affecting cell viability. In addition, phaseolin suppressed pro-inflammatory mediators such as cyclooxygenase 2 (COX-2), interleukin-1β (IL-1β), tumor necrosis factor α (TNF-α), monocyte chemoattractant protein-1 (MCP-1), and interleukin-6 (IL-6) in a dose-dependent manner. Furthermore, phaseolin reduced matrix metalloproteinase (MMP) activity as well as macrophage adhesion in vitro and the recruitment of leukocytes in vivo by downregulating Ninjurin 1 (Ninj1), an adhesion molecule. Finally, phaseolin inhibited the nuclear translocation of nuclear factor-kappa B (NF-κB). In view of the above, our results suggest that phaseolin could be a potential therapeutic candidate for the management of inflammation.

scholarly journals Nutritional Value and Anti-inflammation Activity of Misutkaru with Added Gryllus bimaculatus Powder

2021 ◽  
Vol 19 (3) ◽  
pp. 467-476
Author(s):  
Jung-Soon Han
Keyword(s):  
Amino Acid ◽  
Inflammatory Cytokines ◽  
Nutritional Value ◽  
Interleukin 1 ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Gryllus Bimaculatus ◽  
Necrosis Factor Alpha ◽  
Automatic Amino Acid Analyzer ◽  
Tnf Α

Purpose: This study investigated the nutritional value of Misutkaru with added Gryllus bimaculatus powder (GBM) and its applicability as a healthy functional food.Methods: Chemical analysis of the moisture, crude fat, protein, and mineral contents was performed in accordance with the Association of Official Analytical Chemists (AOAC) guidelines. The amino acid and fatty acid compositions were analyzed using an automatic amino acid analyzer and gas chromatography, respectively. The levels of inflammatory cytokines tumor necrosis factor‑alpha (TNF-α), interleukin 1 beta (IL-1β), and interleukin 6 (IL‑6) induced by lipopolysaccharides in RAW 264.7 cells were measured.Results: The general composition per 100 g of GBM was 41.87 g protein, 19.75 g fat, and 28.52 g carbohydrates. The mineral content per 100 g of GBM was 889.66 mg calcium, 1189.73 mg potassium, 220.36 mg magnesium, 207.51 mg sodium, 694.81 mg phosphorus, and 15.50 mg zinc. In particular, valine (21.361 mg/kg), leucine (29.180 mg/kg), and isoleucine (15.562 mg/kg) were abundant in GBM. GBM also effectively downregulated the production of the inflammatory cytokines TNF-α, IL-1β, and IL-6 in RAW 264.7 macrophages.Conclusion: Misutkaru with added Gryllus bimaculatus powder may have potential for application in the development of food materials or foods to prevent muscle loss in elderly individuals and sarcopenia patients, build muscle, and prevent increase in blood lipid concentrations in middle aged people. In particular, as Gryllus bimaculatus is low in fat and carbohydrates, it can be used as a diet material.

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