Protective Effect of (+)-Catechin Against Lipopolysaccharide Induced Inflammatory Response in RAW 264.7 Cells through Downregulation of NF-κB and p38 MAPK

Abstract Catechin, a flavonol belonging to the flavonoid group of polyphenols is present in many plant foods. The present study was done to evaluate the effect of catechin on various inflammatory mediators using lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The effect of catechin on total cyclooxygenase (COX) activity, 5-lipoxygenase (5-LOX), myeloperoxidase, nitrite and inducible nitric oxide synthase (iNOS) level, secretion of tumour necrosis factor-α (TNF-α) and interleukin-10 (IL-10) were assessed in LPS-stimulated RAW 264.7 cells. The expression of COX-2, iNOS, TNF-α, nuclear factor-ĸB (NF-κB) and p38 mitogen-activated protein kinase (MAPK) genes were also investigated. The effect was further analyzed using human PBMCs by assessing the level of TNF-α and IL-10. The study demonstrated that the inflammatory mediators such as COX, 5-LOX, nitrite, iNOS, and TNF-α were significantly inhibited by catechin in a dose-dependent manner whereas IL-10 production was up-regulated in RAW 264.7 cells. Moreover, catechin down-regulated the mRNA level expression of COX-2, iNOS, TNF-α, NF-κB and p38 MAPK. The current study ratifies the beneficial effect of catechin as a dietary component in plant foods to provide protection against inflammatory diseases.

Download Full-text

Fermentation Improves Anti-Inflammatory Effect of Sipjeondaebotang on LPS-Stimulated RAW 264.7 Cells

The American Journal of Chinese Medicine ◽

10.1142/s0192415x12500619 ◽

2012 ◽

Vol 40 (04) ◽

pp. 813-831 ◽

Cited By ~ 10

Author(s):

You-Chang Oh ◽

Won-Kyung Cho ◽

Yun Hee Jeong ◽

Ga Young Im ◽

Min Cheol Yang ◽

...

Keyword(s):

Nitric Oxide ◽

Inflammatory Mediators ◽

High Performance ◽

Biological Effects ◽

Inhibitory Effect ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Tnf Α ◽

Cox 2 ◽

Anti Inflammatory

Sipjeondaebotang (SJ) has been used as a traditional drug in east-Asian countries. In this study, to provide insight into the biological effects of SJ and SJ fermented by Lactobacillus, we investigated their effects on lipopolysaccharide (LPS)-mediated inflammation in macrophages. The investigation was focused on whether SJ and fermented SJ could inhibit the production of pro-inflammatory mediators such as prostaglandin (PG) E2 and nitric oxide (NO) as well as the expressions of cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS), tumor necrosis factor (TNF)-α, mitogen-activated protein kinases (MAPKs) and nuclear factor (NF)-κB in LPS-stimulated RAW 264.7 cells. We found that SJ modestly inhibited LPS-induced PGE2, NO and TNF-α production as well as the expressions of COX-2 and iNOS. Interestingly, fermentation significantly increased its inhibitory effect on the expression of all pro-inflammatory mediators. Furthermore, fermented SJ exhibited increased inhibition of p38 MAPK and c-Jun NH2-terminal kinase (JNK) MAPK phosphorylation as well as NF-κB p65 translocation by reduced IκBα degradation compared with either untreated controls or unfermented SJ. High performance liquid chromatography (HPLC) analysis showed fermentation by Lactobacillus increases liquiritigenin and cinnamyl alcohol contained in SJ, which are known for their anti-inflammatory activities. Finally, SJ fermented by Lactobacillus exerted potent anti-inflammatory activity by inhibiting MAPK and NF-κB signaling in RAW 264.7 cells.

Download Full-text

Induction of apoptosis by particulate matter: role of TNF-α and MAPK

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1998.275.5.l942 ◽

1998 ◽

Vol 275 (5) ◽

pp. L942-L949 ◽

Cited By ~ 10

Author(s):

Beek Yoke Chin ◽

Mary E. Choi ◽

Marie D. Burdick ◽

Robert M. Strieter ◽

Terence H. Risby ◽

...

Keyword(s):

Particulate Matter ◽

Carbon Black ◽

Mapk Pathway ◽

Time Dependent ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Dependent Manner ◽

Mapk Kinase ◽

Tnf Α ◽

Induced Apoptosis

Particulate matter (PM) is a major by-product from the combustion of fossil fuels. The biological target of inhaled PM is the pulmonary epithelium and resident macrophages. In this study, we demonstrate that cultured macrophages (RAW 264.7 cells) exposed continously to a well-defined model of PM [benzo[ a]pyrene adsorbed on carbon black (CB+BaP)] exhibit a time-dependent expression and release of the cytokine tumor necrosis factor-α (TNF-α). CB+BaP also evoked programmed cell death or apoptosis in cultured macrophages as assessed by genomic DNA-laddering assays. The CB+BaP-induced apoptosis was inhibited when macrophages were treated with CB+BaP in the presence of a neutralizing antibody to TNF-α, suggesting that TNF-α plays an important role in mediating CB+BaP-induced apoptosis in macrophages. Interestingly, neither untreated carbon black nor benzo[ a]pyrene alone induced apoptosis or caused the release of TNF-α in RAW 264.7 cells. Moreover, we observed that TNF-α activates mitogen-activated protein kinase (MAPK) activity, the extracellular signal-regulated kinases p42/p44, in a time-dependent manner. RAW 264.7 cells treated with PD-098059, a selective inhibitor of MAPK kinase activity, did not exhibit CB+BaP-induced apoptosis and TNF-α secretion. Furthermore, cells treated with the MAPK kinase inhibitor did not undergo TNF-α-induced apoptosis. Taken together, our data suggest that TNF-α mediates PM-induced apoptosis and that the MAPK pathway may play an important role in regulating this pathway.

Download Full-text

Bakuchiol Suppresses Inflammatory Responses Via the Downregulation of the p38 MAPK/ERK Signaling Pathway

International Journal of Molecular Sciences ◽

10.3390/ijms20143574 ◽

2019 ◽

Vol 20 (14) ◽

pp. 3574 ◽

Cited By ~ 4

Author(s):

Hye-Sun Lim ◽

Yu Jin Kim ◽

Bu-Yeo Kim ◽

Soo-Jin Jeong

Keyword(s):

Mrna Expression ◽

P38 Mapk ◽

Inflammatory Responses ◽

Mitogen Activated Protein Kinase ◽

Enzyme Linked Immunosorbent Assay ◽

Mice Model ◽

Tnf Α ◽

Cox 2

The purpose of the present study was to evaluate the effects of bakuchiol on the inflammatory response and to identify the molecular mechanism of the inflammatory effects in a lipopolysaccharide (LPS)-stimulated BV-2 mouse microglial cell line and mice model. The production of prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) was measured by enzyme-linked immunosorbent assay. The mRNA expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), TNF-α, and IL-6 was measured using reverse transcription–polymerase chain reaction analysis. Mitogen-activated protein kinase (MAPK) phosphorylation was determined by western blot analysis. In vitro experiments, bakuchiol significantly suppressed the production of PGE2 and IL-6 in LPS-stimulated BV-2 cells, without causing cytotoxicity. In parallel, bakuchiol significantly inhibited the LPS-stimulated expression of iNOS, COX-2, and IL-6 in BV-2 cells. However, bakuchiol had no effect on the LPS-stimulated production and mRNA expression of TNF-α or on LPS-stimulated c-Jun NH2-terminal kinase phosphorylation. In contrast, p38 MAPK and extracellular signal-regulated kinase (ERK) phosphorylation were inhibited by bakuchiol. In vivo experiments, Bakuchiol reduced microglial activation in the hippocampus and cortex tissue of LPS-injected mice. Bakuchiol significantly suppressed LPS-injected production of TNF-α and IL-6 in serum. These results indicate that the anti-neuroinflammatory effects of bakuchiol in activated microglia are mainly regulated by the inhibition of the p38 MAPK and ERK pathways. We suggest that bakuchiol may be beneficial for various neuroinflammatory diseases.

Download Full-text

Inhibitory Effect of 1,5-Dimethyl Citrate from Sea Buckthorn (Hippophae rhamnoides) on Lipopolysaccharide-Induced Inflammatory Response in RAW 264.7 Mouse Macrophages

Foods ◽

10.3390/foods9030269 ◽

2020 ◽

Vol 9 (3) ◽

pp. 269 ◽

Cited By ~ 1

Author(s):

Su Cheol Baek ◽

Dahae Lee ◽

Mun Seok Jo ◽

Kwang Ho Lee ◽

Yong Hoon Lee ◽

...

Keyword(s):

Nitric Oxide ◽

Nitric Oxide Synthase ◽

Sea Buckthorn ◽

Raw 264.7 Cells ◽

No Production ◽

Raw 264.7 ◽

Tnf Α ◽

Mouse Macrophages ◽

Cox 2 ◽

Oxide Synthase

Hippophae rhamnoides L. (Elaeagnaceae; commonly known as “sea buckthorn” and “vitamin tree”), is a spiny deciduous shrub whose fruit is used in foods and traditional medicines. The H. rhamnoides fruit (berry) is rich in vitamin C, with a level exceeding that found in lemons and oranges. H. rhamnoides berries are usually washed and pressed to create pomace and juice. Today, the powder of the aqueous extract of H. rhamnoides berries are sold as a functional food in many countries. As part of our ongoing effort to identify bioactive constituents from natural resources, we aimed to isolate and identify those from the fruits of H. rhamnoides. Phytochemical analysis of the extract of H. rhamnoides fruits led to the isolation and identification of six compounds, namely, a citric acid derivative (1), a phenolic (2), flavonoids (3 and 4), and megastigmane compounds (5 and 6). Treatment with compounds 1–6 did not have any impact on the cell viability of RAW 264.7 mouse macrophages. However, pretreatment with these compounds suppressed lipopolysaccharide (LPS)-induced NO production in RAW 264.7 mouse macrophages in a concentration-dependent manner. Among the isolated compounds, compound 1 was identified as the most active, with an IC50 of 39.76 ± 0.16 μM. This value was comparable to that of the NG-methyl-L-arginine acetate salt, a nitric oxide synthase inhibitor with an IC50 of 28.48 ± 0.05 μM. Western blot analysis demonstrated that compound 1 inhibited the LPS-induced expression of IKKα/β (IκB kinase alpha/beta), I-κBα (inhibitor of kappa B alpha), nuclear factor kappa-B (NF-κB) p65, iNOS (inducible nitric oxide synthase), and COX-2 (cyclooxygenase-2) in RAW 264.7 cells. Furthermore, LPS-stimulated cytokine production was detected using a sandwich enzyme-linked immunosorbent assay. Compound 1 decreased interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α) production in LPS-stimulated RAW 264.7 cells. In summary, the mechanism of action of 1 included the suppression of LPS-induced NO production in RAW 264.7 cells by inhibiting IKKα/β, I-κBα, NF-κB p65, iNOS, and COX-2, and the activities of IL-6 and TNF-α.

Download Full-text

Anti-inflammatory Effects of Achillea millefolium on Atopic Dermatitis-Like Skin Lesions in NC/Nga Mice

The American Journal of Chinese Medicine ◽

10.1142/s0192415x2050055x ◽

2020 ◽

Vol 48 (05) ◽

pp. 1121-1140

Author(s):

Hien T.T. Ngo ◽

Eunson Hwang ◽

Hyungoo Kang ◽

Bom Park ◽

Seul A. Seo ◽

...

Keyword(s):

Atopic Dermatitis ◽

Elevated Serum ◽

Mitogen Activated Protein Kinase ◽

Raw 264.7 Cells ◽

Achillea Millefolium ◽

Raw 264.7 ◽

Hacat Cells ◽

Necrosis Factor Alpha ◽

Cox 2 ◽

Anti Inflammatory

Achillea millefolium L. (AM) is an aromatic herb with a variety of pharmacological properties, such as anti-inflammatory and anti-allergic activities. However, AM’s effects on atopic dermatitis (AD) have not been investigated. This study evaluates the anti-AD activity of 50% ethanol-extracted AM in murine macrophage Raw 264.7 cells, in tumor necrosis factor-alpha/interferon-gamma (TNF-[Formula: see text]/IFN-[Formula: see text])-stimulated human immortal keratinocyte HaCaT cells in vitro, and in Biostir-AD-treated NC/Nga mice in vivo. The results showed that AM significantly downregulated expression of pro-inflammatory cytokines, such as INOS, COX-2, and interleukin (IL)-6 in lipopolysaccharide (LPS)-treated Raw 264.7 cells. The mRNA expressions of INOS, COX-2, and IL-6 decreased by 76.1%, 69.3%, and 31.8%, respectively. Overexpression of chemokines, such as activation-regulated chemokine and macrophage-derived chemokine, regulated on activation of normal T-cell expressed and secreted, and IL-8 was inhibited by 70.01%, 52.91%, 73.53%, and 18.93%, respectively, in TNF-[Formula: see text]/IFN-[Formula: see text]-stimulated HaCaT cells by downregulating the mitogen-activated protein kinase, I[Formula: see text]B[Formula: see text], and the signal transducer and activator of transcription 1 signaling pathways. AD-like symptoms, such as elevated serum immunoglobin E levels, epidermal thickening, high dermatitis severity score, transepidermal water loss, and reduced skin hydration, were relieved by the dietary administration of AM in Biostir-AD-treated NC/Nga mice. In addition, filaggrin expression increased significantly in AM-treated groups. These results suggest that AM could be a useful candidate for AD treatment.

Download Full-text

The Immunomodulatory Activities of Picria Fel-Terrae Lour Herbs towards RAW 264.7 Cells

Open Access Macedonian Journal of Medical Sciences ◽

10.3889/oamjms.2019.017 ◽

2019 ◽

Vol 7 (1) ◽

pp. 24-28

Author(s):

Novycha AuliaFendri ◽

Rosidah ◽

Yuandani ◽

Sri Suryani ◽

Denny Satria

Keyword(s):

Ethyl Acetate ◽

Inflammatory Biomarkers ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Chromatography Method ◽

Tnf Α ◽

Gene Level ◽

Pcr Method ◽

Cox 2 ◽

Oxide Synthase

AIM: To investigate immunomodulatory activities of Picria fel-terrae Lour herbs extract against inflammatory biomarkers by conducting cell culture experiments. MATERIAL AND METHODS: The herbs of Picria fel-terrae Lour were dried and extracted with n-hexane, ethyl acetate, 96% ethanol, followed by evaporation and freeze-drying. Phytochemicals screening were analysed with thin layer chromatography method. Cell viability was assessed with MTT assay. The genes of Tumor Necrosis Factor (TNF)-α, Interleukin (IL)-6, interleukin (IL)-1β and inducible Nitric Oxide Synthase (iNOS), Cyclooxygenase (COX-2) in lipopolysaccharide (LPS)-induced macrophages were analysed by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) method. RESULTS: Phytochemicals screening showed the presence of steroids in n-hexane extract (ENPFH) and flavonoids, glycosides, saponins, tannins in ethyl acetate (EEAPFH) and ethanol (EEPFH) extracts. The Viability of RAW 264.7 cell toward ENPFH, EEAPFH, and EEPFH (1-200 μgmL-1) showed no toxicity effects. At the gene level, ENPFH; EEAPFH; EEPFH decreased the gene expression of TNF-α, IL-6, IL-1β, iNOS, and COX-2 which induced with LPS (1 μgmL-1). CONCLUSIONS: Our results suggest that extracts of Picria fel-terrae Lour Herbs possesses immunomodulatory activities by inhibiting selected inflammatory biomarkers at the gene levels in LPS-induced macrophages.

Download Full-text

Dilodendron bipinnatum Radlk. inhibits pro-inflammatory mediators through the induction of MKP-1 and the down-regulation of MAPKp38/JNK/NF-κB pathways and COX-2 in LPS-activated RAW 264.7 cells

Journal of Ethnopharmacology ◽

10.1016/j.jep.2017.02.026 ◽

2017 ◽

Vol 202 ◽

pp. 127-137 ◽

Cited By ~ 9

Author(s):

Ruberlei Godinho de Oliveira ◽

Geovane Roberto de Campos Castilho ◽

André Luiz da Cunha ◽

Fábio Miyajima ◽

Domingos Tabajara de Oliveira Martins

Keyword(s):

Inflammatory Mediators ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Down Regulation ◽

Cox 2

Download Full-text

Anti-inflammatory activity of pectic enzyme-treated pectin on lipopolysaccharide-induced RAW 264.7 cells

Journal of Functional Food and Nutraceutical ◽

10.33555/jffn.v1i1.14 ◽

2019 ◽

Vol 1 (1) ◽

pp. 23-30 ◽

Cited By ~ 1

Author(s):

Cian-Song Huang ◽

Qiao-Lin Li ◽

Diana Lo ◽

Yuh-Tai Wang ◽

Ming-Chang Wu

Keyword(s):

Western Blot ◽

Raw 264.7 Cells ◽

No Production ◽

Raw 264.7 ◽

Pectic Enzyme ◽

Western Blot Method ◽

Tnf Α ◽

Cox 2 ◽

Necrosis Factor ◽

Normal Macrophage

The purpose of this study was to investigate the ability and pathway of the pectic enzyme-treated (PET) pectin to inhibit the inflammation of macrophage RAW 264.7 induced by lipopolysaccharide. Results showed that PET-pectin produced from 1% substrate and 48 h reaction time had the highest antioxidative activity, thus these parameters were used to produce PET-pectin used in this study. PET-pectin showed no cell cytotoxicity to normal macrophage RAW 264.7 and reduce the nitrite secretion from LPS-induced RAW 264.7 by 20%. Finally, the expression of cytokines, including NO synthase (iNOS), nitric oxide (NO), cyclooxygenase-2 (COX-2), nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and tumor necrosis factor (TNF-α) were analyzed by western blot. In the western blot method, it was found that iNOS, COX-2, NF-κB, TNF-α and other proteins that activated NO production had a downtrend. It was found that PET-pectin possess promising activity to mitigate the inflammatory response.

Download Full-text

Effects of Sibseonsan as an Anti-Inflammatory, Anti-Wrinkle, and Skin Whitening Treatment

Journal of Acupuncture Research ◽

10.13045/jar.2020.00024 ◽

2020 ◽

Vol 37 (2) ◽

pp. 88-93

Author(s):

Na Young Jo

Keyword(s):

Treatment Group ◽

Cell Line ◽

Radical Scavenging ◽

Raw 264.7 Cells ◽

No Production ◽

Raw 264.7 ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Tnf Α ◽

Anti Inflammatory

Background: The purpose of this study was to investigate whether Sibseonsan (SSS) is an effective anti-inflammatory, anti-wrinkling, and whitening agent.Methods: To determine whether SSS had an anti-inflammatory effect, a murine macrophage cell line was used (RAW 264.7) and production of DPPH, NO, TNF-α, and PGE2 were measured. To ascertain potential anti-wrinkle effects of SSS in these cells, collagenase and elastase production were measured. To verify whether SSS had a whitening effect, tyrosinase activity and DOPA staining were performed using a melanoma cell line (B16/F10).Results: There was no significant reduction in survival of SSS-treated RAW 264.7 cells, up to 400 μg/mL. Free radical scavenging (23.96 ± 1.85%) was observed in RAW 264.7 cells treated with SSS at a concentration of 400 μg/mL. The SSS treatment group (400 μg/mL) significantly inhibited NO production compared with the LPS stimulated treatment group. The SSS treatment of macrophage cells appeared to reduce production of TNF-α in a concentration dependent manner. There was a significant reduction in the concentration of PGE2 by about 25% in the SSS treatment (400 μg/mL) group (p = 0.05). Compared with the control, the production of collagenase and elastase in B16/F10 cells treated with SSS (400 μg/mL) was greater by 26.37% and 45.71%, respectively. The SSS treatment (400 μg/mL) group showed a significant reduction by about 17% in tyrosinase production in B16/F10 cells. The SSS treatment group showed little change in DOPA staining. Conclusion: SSS extract may be useful for the treatment and prevention of inflammatory diseases and may have anti-wrinkle and whitening effects. These results may support the use of SSS in clinical practice.

Download Full-text