Agrimonia pilosa Ledeb Root Extract: Anti-Inflammatory Activities of the Medicinal Herb in LPS-Induced Inflammation

Inflammation regulation is essential for maintaining healthy functions and normal homeostasis of the body. Porphyromonas gingivalis (P. gingivalis) is a gram-negative anaerobic bacterium and a major pathogen that causes oral inflammation and other systemic inflammations. This study aims to examine the anti-inflammatory effects of Agrimonia pilosa Ledeb root extracts (APL-ME) in Porphyromonas gingivalis LPS-induced RAW 264.7 cells and find anti-inflammatory effect compounds of APL-ME. The anti-inflammatory effects of APL-ME were evaluated anti-oxidant activity, cell viability, nitrite concentration, pro-inflammatory cytokines (interleukin-1[Formula: see text], interleukin-6, tumor necrosis factor (TNF)-[Formula: see text], and anti-inflammatory cytokine (interleukin-10 (IL-10)). Also, Inflammation related genes and proteins, cyclooxygenase (COX)-2, inducible nitric oxide synthase (iNOS), expression were decreased by APL-ME and mitogen-activated protein kinase (MAPK) signaling proteins expression was regulated by APL-ME. Liquid chromatography-mass spectrometer (LC/MS)-MS analysis results indicated that several components were detected in APL-ME. Our study indicated that APL-ME suppressed nitrite concentrations, pro-inflammatory cytokines such as IL-1[Formula: see text], IL-6 and TNF-[Formula: see text] in P. gingivalis LPS induced RAW 264.7 cells. However, IL-10 expression was increased by ALP-ME. In addition, protein expressions of COX-2 and iNOS were inhibited APL-ME extracts dose-dependently. According to these results, APL-ME has anti-inflammatory effects in P. gingivalis LPS induced RAW 264.7 cells.

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Anti-Inflammatory Effects of 6,7-Dihydroxy-4-Methylcoumarin on LPS-Stimulated Macrophage Phosphorylation in MAPK Signaling Pathways

Natural Product Communications ◽

10.1177/1934578x211020970 ◽

2021 ◽

Vol 16 (5) ◽

pp. 1934578X2110209

Author(s):

Yun Sil Kang ◽

You Chul Chung ◽

Jung No Lee ◽

Bong Seok Kim ◽

Chang-Gu Hyun

Keyword(s):

Signaling Pathways ◽

Inflammatory Cytokines ◽

Inflammatory Diseases ◽

Mapk Signaling ◽

Raw 264.7 Cells ◽

Inflammatory Factors ◽

Raw 264.7 ◽

Dependent Manner ◽

Anti Inflammatory ◽

Pro Inflammatory Cytokines

Coumarin derivatives, such as esculetin, have various physiological functions, including antioxidant, anti-inflammatory, antibacterial, antiviral, and anti-cancer. 6,7-Dihydroxy-4-methylcoumarin (6,7-DH-4MC) is a derivative of esculetin, and its anti-inflammatory effect and mechanism in macrophages have not been studied. In this study, the anti-inflammatory activity of 6,7-DH-4MC was evaluated by measuring the expression of inflammatory factors (NO and PGE2) and pro-inflammatory cytokines (IL-1β, IL-6, and TNF-α) in LPS-stimulated RAW 264.7 macrophages. The results revealed that 6,7-DH-4MC significantly reduced NO levels and PGE2 expression without inducing cytotoxicity; it was confirmed that the inhibition of NO and PGE2 expression was related to iNOS and COX-2 downregulation in response to 6,7-DH-4MC treatment. Moreover, 6,7-DH-4MC decreased the levels of pro-inflammatory cytokines, such as IL-1β and IL-6, in a dose-dependent manner. Mechanistic studies revealed reduced phosphorylation of ERK and p38-MAPK upon 6,7-DH-4MC treatment. Furthermore, the degradation of IκB-α and phosphorylation of NF-κB in cells treated with LPS were interrupted by 6,7-DH-4MC treatment. These results suggest that 6,7-DH-4MC is a potential therapeutic agent for inflammatory diseases. To the best of our knowledge, this is the first report demonstrating the anti-inflammatory effects of 6,7-DH-4MC in RAW 264.7 cells via MAPK and NF-κB signaling pathways.

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Anti-Inflammatory Effects of 6-Methylcoumarin in LPS-Stimulated RAW 264.7 Macrophages via Regulation of MAPK and NF-κB Signaling Pathways

Molecules ◽

10.3390/molecules26175351 ◽

2021 ◽

Vol 26 (17) ◽

pp. 5351

Author(s):

Jin-Kyu Kang ◽

You-Chul Chung ◽

Chang-Gu Hyun

Keyword(s):

Nitric Oxide ◽

Inflammatory Cytokines ◽

Mitogen Activated Protein Kinase ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Anti Inflammatory ◽

Pro Inflammatory Cytokines

Persistent inflammatory reactions promote mucosal damage and cause dysfunction, such as pain, swelling, seizures, and fever. Therefore, in this study, in order to explore the anti-inflammatory effect of 6-methylcoumarin (6-MC) and suggest its availability, macrophages were stimulated with lipopolysaccharide (LPS) to conduct an in vitro experiment. The effects of 6-MC on the production and levels of pro-inflammatory cytokines (interleukin (IL)-1β, IL-6, tumor necrosis factor (TNF)-α) and inflammatory mediators (nitric oxide (NO), prostaglandin E2 (PGE2)) in LPS-stimulated RAW 264.7 cells were examined. The results showed that 6-MC reduced the levels of NO and PGE2 without being cytotoxic. In addition, it was demonstrated that the increase in the expression of pro-inflammatory cytokines caused by LPS stimulation, was decreased in a concentration-dependent manner with 6-MC treatment. Moreover, Western blot results showed that the protein levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), which increased with LPS treatment, were decreased by 6-MC treatment. Mechanistic studies revealed that 6-MC reduced the phosphorylation of the mitogen-activated protein kinase (MAPK) family and IκBα in the MAPK and nuclear factor-kappa B (NF-κB) pathways, respectively. These results suggest that 6-MC is a potential therapeutic agent for inflammatory diseases that inhibits inflammation via the MAPK and NF-κB pathways.

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Decursinol Angelate Inhibits LPS-Induced Macrophage Polarization through Modulation of the NFκB and MAPK Signaling Pathways

Molecules ◽

10.3390/molecules23081880 ◽

2018 ◽

Vol 23 (8) ◽

pp. 1880 ◽

Cited By ~ 15

Author(s):

Salman Islam ◽

Jung Lee ◽

Adeeb Shehzad ◽

Eun-Mi Ahn ◽

You Lee ◽

...

Keyword(s):

Map Kinase ◽

Signaling Pathways ◽

Inflammatory Cytokines ◽

Macrophage Polarization ◽

Raw 264.7 Cells ◽

M2 Macrophage ◽

Raw 264.7 ◽

Decursinol Angelate ◽

Anti Inflammatory ◽

Pro Inflammatory Cytokines

Inflammation is considered the root cause of various inflammatory diseases, including cancers. Decursinol angelate (DA), a pyranocoumarin compound obtained from the roots of Angelica gigas, has been reported to exhibit potent anti-inflammatory effects. In this study, the anti-inflammatory effects of DA on the MAP kinase and NFκB signaling pathways and the expression of pro-inflammatory cytokines were investigated in phorbol 12-myristate 13-acetate (PMA)-activated human promyelocytic leukemia (HL-60) and lipopolysaccharide (LPS)-stimulated macrophage (Raw 264.7) cell lines. PMA induced the activation of the MAP kinase-NFκB pathway and the production of pro-inflammatory cytokines in differentiated monocytes. Treatment with DA inhibited the activation of MAP kinases and the translocation of NFκB, and decreased the expression and exogenous secretion of IL-1β and IL-6. Furthermore, LPS-stimulated Raw 264.7 cells were found to have increased expression of M1 macrophage-associated markers, such as NADPH oxidase (NOX) and inducible nitric oxide synthase (iNOS), and the M2 macrophage-associated marker CD11b. LPS also activated pro-inflammatory cytokines and Erk-NFκB. Treatment with DA suppressed LPS-induced macrophage polarization and the inflammatory response by blocking Raf-ERK and the translocation of NFκB in Raw 264.7 cells. Treatment with DA also inhibited the expression of pro-inflammatory cytokines, such as IL-1β and IL-6, NOX, and iNOS in Raw 264.7 cells. These results suggest that DA has the potential to inhibit macrophage polarization and inflammation by blocking the activation of pro-inflammatory signals. These anti-inflammatory effects of DA may contribute to its potential use as a therapeutic strategy against various inflammation-induced cancers.

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5-Hydroxymethylfurfural Mitigates Lipopolysaccharide-Stimulated Inflammation via Suppression of MAPK, NF-κB and mTOR Activation in RAW 264.7 Cells

Molecules ◽

10.3390/molecules24020275 ◽

2019 ◽

Vol 24 (2) ◽

pp. 275 ◽

Cited By ~ 13

Author(s):

Fanhui Kong ◽

Bae Lee ◽

Kun Wei

Keyword(s):

Nitric Oxide ◽

Inflammatory Cytokines ◽

Mammalian Target Of Rapamycin ◽

Inflammatory Activity ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Dependent Manner ◽

Concentration Dependent Manner ◽

Anti Inflammatory ◽

Pro Inflammatory Cytokines

5-Hydroxymethylfurfural (5-HMF) is found in many food products including honey, dried fruits, coffee and black garlic extracts. Here, we investigated the anti-inflammatory activity of 5-HMF and its underlying mechanisms in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. 5-HMF pretreatment ranging from 31.5 to 126.0 μg/mL reduced the production of nitric oxide (NO), prostaglandin E2 (PGE2) and pro-inflammatory cytokines (TNF-α, IL-1β and IL-6) in a concentration-dependent manner in LPS-stimulated cells. Moreover, 5-HMF-pretreated cells significantly down-regulated the mRNA expression of two major inflammatory mediators, nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) and suppressed the production of pro-inflammatory cytokines, as compared with the only LPS-stimulated cells. 5-HMF suppressed the phosphorylation of extracellular regulated protein kinases (ERK1/2), c-Jun N-terminal kinase (JNK), IκBα, NF-κB p65, the mammalian target of rapamycin (mTOR) and protein kinase B (Akt). Besides, 5-HMF was proved to inhibit NF-κB p65 translocation into nucleus to activate inflammatory gene transcription. These results suggest that 5-HMF could exert the anti-inflammatory activity in the LPS-induced inflammatory response by inhibiting the MAPK, NF-κB and Akt/mTOR pathways. Thus, 5-HMF could be considered as a therapeutic ingredient in functional foods.

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Effect of Fractions, and Compounds from Typha capensis in LPS-Stimulated Raw 264.7 Cells. Pro and Anti-inflammatory Cytokines

Current Journal of Applied Science and Technology ◽

10.9734/cjast/2021/v40i1431399 ◽

2021 ◽

pp. 20-27

Author(s):

Moise Ondua

Keyword(s):

Inflammatory Cytokines ◽

Interleukin 1 ◽

Molecular Targets ◽

Traditional Healers ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Necrosis Factor Alpha ◽

Tnf Α ◽

Anti Inflammatory ◽

Pro Inflammatory Cytokines

Typha capensis is widely used by traditional healers to treat male fertility, venereal problems and inflammation. There are many molecular targets implicated in the inflammatory process: pro- and anti-inflammatory cytokines such as interleukin 1-β, IL-6, IL-10, IL-12p70, tumor necrosis factor alpha (TNF-α), and IL-8, and other proteins such as COX-2, and iNOS. In order to clarify the anti-inflammatory mechanism of action of compounds isolated from T. capensis, RAW 264.7 macrophages were activated by lipopolysaccharide and pre-treated with T. capensis isolated compounds. Lipopolysaccharide-stimulated RAW macrophages after treatment with T. capensis crude acetone extract resulted in decreasing expression of pro-inflammatory cytokines (TNF-α, IL-6,) and increased expression of immunomodulatory cytokine IL-12 P 70. Isorhamnetin-3-O-β-D-glucoside and isorhamnetin 3-O rutinoside increased the expression of pro-inflammatory cytokines TNF-α, but failed to reduce the expression of IL-1β and TNF-α. Isorhamnetin-3-O-β-D-glucoside and isorhamnetin 3-O rutinoside increased the expression of immunomodulatory cytokine IL-12p70. Isorhamnetin-3-O-β-D-glucoside increased the expression of the anti-inflammatory cytokine IL-10 compared to quercetin and LPS-stimulated macrophages. The effect of isorhamnetin 3-O-rutinoside and isorhamnetin-3-O-β-D-glucoside on molecular targets of inflammation may provide support for the use of T. capensis by traditional healers against inflammation.

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Scandoside Exerts Anti-Inflammatory Effect via Suppressing NF-κB and MAPK Signaling Pathways in LPS-Induced RAW 264.7 Macrophages

10.20944/preprints201801.0170.v1 ◽

2018 ◽

Author(s):

Jingyu He ◽

Jiafeng Li ◽

Han Liu ◽

Zichao Yang ◽

Fenghua Zhou ◽

...

Keyword(s):

Nitric Oxide ◽

Transcription Factor ◽

Mapk Signaling ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Nuclear Transcription Factor ◽

Cox 2 ◽

Anti Inflammatory ◽

Mapk Signaling Pathways ◽

Inflammatory Effect

The iridoids of H. diffusa play an important role in the anti-inflammatory process, but the specific iridoid with anti-inflammatory effect and its mechanism is lack of study. An iridoid compound named scandoside (SCA) was isolated from H. diffusa and its anti-inflammatory effect was investigated in lipopolysaccharide (LPS)-induced RAW 264.7 cells. Its anti-inflammatory mechanism was confirmed by in intro experiment and molecular docking analysis. As results, SCA significantly decreased the productions of nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) and inhibited the levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), TNF-α and IL-6 mRNA expression in LPS-induced RAW 264.7 cells. SCA treatment suppressed the phosphorylation of inhibitor of nuclear transcription factor kappa-B alpaha (IκB-α), p38, extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK). The docking data suggested that SCA had great binding abilities to COX-2, iNOS and IκB. Taken together, the results indicated that the anti-inflammatory effect of SCA is due to inhibition of pro-inflammatory cytokines and mediators via suppressing the nuclear transcription factor kappa-B (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways, which provided useful information for its application and development.

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Baicalin improves the survival in endotoxic mice and inhibits the inflammatory responses in LPS-treated RAW 264.7 macrophages

European Journal of Inflammation ◽

10.1177/2058739220967767 ◽

2020 ◽

Vol 18 ◽

pp. 205873922096776

Author(s):

Shi-Wen Kuo ◽

Wen-Lin Su ◽

Tz-Chong Chou

Keyword(s):

Survival Rate ◽

Inflammatory Cytokines ◽

Nuclear Translocation ◽

High Morbidity ◽

Raw 264.7 ◽

Myeloperoxidase Activity ◽

Raw 264.7 Macrophages ◽

Cox 2 ◽

Anti Inflammatory ◽

Pro Inflammatory Cytokines

Introduction: Sepsis is a severe disease with a high morbidity and mortality. Baicalin, an active compound of Chinese medicine, Scutellaria baicalensis Georgi (Huang Qui), exhibits several beneficial effects. In this study, we examined whether administration of baicalin increases the survival in mice with endotoxemia and investigated its anti-inflammatory mechanisms in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Methods: The production of NOx, PGE2, and pro-inflammatory cytokines, the mRNA and protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), and the nuclear translocation of NF-κB in LPS-stimulated macrophages or endotoxic mice were determined. The model of severe endotoxic mice was established by injection of LPS (60 mg/kg, i.p.). Results: Baicalin significantly inhibited the production of NO, PGE2, and pro-inflammatory cytokines, including TNF-α, IL-1β, and IL-6 in LPS-stimulated macrophages. Baicalin treatment also markedly suppressed LPS-induced iNOS and COX-2 expression at the transcriptional and translational levels, and the nuclear translocation of NF-κB in macrophages. Similarly, the serum concentrations of NOx, PGE2, and pro-inflammatory cytokines, and the lung myeloperoxidase activity were greatly reduced in baicalin-treated endotoxic mice. Notably, after LPS injection, the 3-day survival rate of mice treated with pre- or post-administration of baicalin (50 mg/kg, i.p.) remarkably increased to 100% and 90%, respectively compared with LPS-injected alone mice with a survival rate of 0%. Conclusion: Baicalin has a potent anti-inflammatory activity in LPS-stimulated macrophages and endotoxic mice. Moreover, treatment with baicalin dramatically increased the survival in the severe septic mice, suggesting that baicalin may be a potential agent for sepsis therapy.

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The Extraction Solvent Influences the Anti-Inflammatory Effects of Jakyakgamcho-Tang in Lipopolysaccharide-Stimulated Macrophages and Mice with Gouty Arthritis

International Journal of Molecular Sciences ◽

10.3390/ijms21249748 ◽

2020 ◽

Vol 21 (24) ◽

pp. 9748

Author(s):

Yun Mi Lee ◽

Dong-Seon Kim

Keyword(s):

Inflammatory Cytokines ◽

Weight Bearing ◽

Gouty Arthritis ◽

Raw 264.7 Cells ◽

Mrna Levels ◽

Extraction Solvent ◽

Cox 2 ◽

Anti Inflammatory ◽

Pg E2 ◽

Pro Inflammatory Cytokines

Jakyakgamcho-Tang (JGT) is a traditional medicine used to treat muscular tension, spasm, and pain. Several studies have reported its clinical use as an anti-inflammatory and in gynaecological treatment. This study aimed to compare the anti-inflammatory effects of JGT according to extraction solvent, water (JGTW) and 30% EtOH (JGTE) on lipopolysaccharide (LPS)—stimulated macrophages and in mice with monosodium urate (MSU)—induced gouty arthritis. We evaluated the production of inflammatory mediators and cytokines and the expression of inducible nitric oxide (iNOS) and cyclooxygenase-2 (COX-2) in RAW 264.7 cells. We also examined oedema, pain, and inflammation in MSU-induced mice by measuring affected hind paw swelling, weight-bearing, pro-inflammatory cytokines levels, and myeloperoxidase (MPO) activity. In LPS-stimulated RAW264.7 cells, JGTW and JGTE significantly decreased prostaglandin (PG) E2(PGE2) production via suppressing COX-2 expression and cytokines interleukin-1β and interleukin-6. Only JGTE reduced the production of NO and cytokines and the mRNA levels of iNOS and cytokines. In MSU-induced mice, JGTE and JGTW efficiently decreased paw swelling and attenuated joint pain. JGTE (200 and 300 mg/kg) effectively suppressed inflammation by downregulating pro-inflammatory cytokines (tumour necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6) and MPO activity, which were only slightly reduced by JGTW. Our data demonstrate the anti-inflammatory activity of JGT in macrophage and gouty arthritis animal models and show that JGTE is more effective than JGTW at lower concentrations.

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Protective Effect of Membrane-Free Stem Cells against Lipopolysaccharide and Interferon-Gamma-Stimulated Inflammatory Responses in RAW 264.7 Macrophages

International Journal of Molecular Sciences ◽

10.3390/ijms22136894 ◽

2021 ◽

Vol 22 (13) ◽

pp. 6894

Author(s):

Mei Tong He ◽

Hye Sook Park ◽

Young Sil Kim ◽

Ah Young Lee ◽

Eun Ju Cho

Keyword(s):

Nitric Oxide ◽

Stem Cells ◽

Interferon Gamma ◽

Mapk Signaling ◽

Raw 264.7 ◽

Raw 264.7 Macrophages ◽

Protein Expressions ◽

Ifn Γ ◽

Cox 2 ◽

Anti Inflammatory

Recently, adipose-derived stem cells (ADSCs) are considered to be ideal for application in cell therapy or tissue regeneration, mainly due to their wide availability and easy access. In this study, we examined the anti-inflammatory effects of membrane-free stem cell extract (MFSC-Ex) derived from ADSCs against lipopolysaccharide (LPS)/interferon-gamma (IFN-γ) on RAW 264.7 macrophage cells. Exposure of RAW macrophages to LPS and IFN-γ stimuli induced high levels of nitric oxide (NO), cyclooxygenase-2 (COX-2), and prostaglandin E2 (PGE2) production. However, pretreatment with MFSC-Ex inhibited LPS/IFN-γ-induced these pro-inflammatory mediators. To clarify the molecular mechanisms underlying the anti-inflammatory property of MFSC-Ex, we analyzed nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinases (MAPKs) protein expressions by Western blotting. Our study showed that treatment of MFSC-Ex significantly down-regulated inducible nitric oxide synthase (iNOS) and COX-2 protein expressions. Furthermore, phosphorylation of extracellular signal-regulated kinase (ERK) and p38 was also blocked by treatment with MFSC-Ex, indicating that inhibitory effect of MFSC-Ex on MAPK signaling cascade may attribute to inactivation of NF-κB. From these findings, we suggest that MFSC-Ex exert anti-inflammatory activities, which suppressed LPS/IFN-γ-induced production of NO, COX-2 and PGE2 by regulation of NF-κB and MAPK signaling pathway in RAW 264.7 macrophages. In conclusion, MFSC-Ex might provide a new therapeutic opportunity to treatment of inflammatory-related diseases.

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