Regulation of Proinflammatory Mediators via NF-κB and p38 MAPK-Dependent Mechanisms in RAW 264.7 Macrophages by Polyphenol Components Isolated from KoreaLonicera japonica THUNB

Lonicera japonica THUNB., which abundantly contains polyphenols, has been used as a traditional medicine for thousands of years in East Asian countries because of the anti-inflammation properties. This study aimed to investigate the anti-inflammatory mechanism of polyphenol components isolated from KoreaL. japonica T.by nuclear factor-kappaB (NF-κB) and mitogen-activated protein kinases (MAPKs) pathway. Polyphenols significantly decreased lipopolysaccharide- (LPS-) induced mRNA and protein expression of inducible nitric oxide synthase and cyclooxygenase-2, as well as mRNA expression of tumor necrosis factor-alpha, interleukin- (IL-) 1β, and IL-6. Moreover, polyphenols inhibited nuclear translocation of NF-κB p65, phosphorylation/degradation of the inhibitor ofκB, and phosphorylation of p38 MAPK, whereas the extracellular signal-regulated kinase and Janus N-terminal kinase were not affected. These results indicate that polyphenol components isolated from KoreaL. japonica T.should have anti-inflammatory effect on LPS-stimulated RAW 264.7 cells through the decrease of proinflammatory mediators expression by suppressing NF-κB and p38 MAPK activity.

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Anti-Inflammatory Effects of Cumin Essential Oil by Blocking JNK, ERK, and NF-κB Signaling Pathways in LPS-Stimulated RAW 264.7 Cells

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2015/474509 ◽

2015 ◽

Vol 2015 ◽

pp. 1-8 ◽

Cited By ~ 12

Author(s):

Juan Wei ◽

Xitong Zhang ◽

Yang Bi ◽

Ruidong Miao ◽

Zhong Zhang ◽

...

Keyword(s):

Essential Oil ◽

Transcriptional Activation ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Extracellular Signal ◽

Mitogen Activated Protein ◽

Anti Inflammatory ◽

Underlying Mechanisms ◽

Oxide Synthase ◽

Inducible Nitric Oxide

Cumin seeds (Cuminum cyminumL.) have been commonly used in food flavoring and perfumery. In this study, cumin essential oil (CuEO) extracted from seeds was employed to investigate the anti-inflammatory effects in lipopolysaccharide- (LPS-) stimulated RAW 264.7 cells and the underlying mechanisms. A total of 26 volatile constituents were identified in CuEO by GC-MS, and the most abundant constituent was cuminaldehyde (48.773%). Mitochondrial-respiration-dependent 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium (MTT) reduction assay demonstrated that CuEO did not exhibit any cytotoxic effect at the employed concentrations (0.0005–0.01%). Real-time PCR tests showed that CuEO significantly inhibited the mRNA expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX-2), interleukin- (IL-) 1, and IL-6. Moreover, western blotting analysis revealed that CuEO blocked LPS-induced transcriptional activation of nuclear factor-kappa B (NF-κB) and inhibited the phosphorylation of extracellular signal regulated kinase (ERK) and c-Jun N-terminal kinase (JNK). These results suggested that CuEO exerted anti-inflammatory effects in LPS-stimulated RAW 264.7 cells via inhibition of NF-κB and mitogen-activated protein kinases ERK and JNK signaling; the chemical could be used as a source of anti-inflammatory agents as well as dietary complement for health promotion.

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Flavonoids Identified from KoreanScutellaria baicalensisGeorgi Inhibit Inflammatory Signaling by Suppressing Activation of NF-κB and MAPK in RAW 264.7 Cells

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2013/912031 ◽

2013 ◽

Vol 2013 ◽

pp. 1-11 ◽

Cited By ~ 17

Author(s):

Gyeong-Eun Hong ◽

Jin-A. Kim ◽

Arulkumar Nagappan ◽

Silvia Yumnam ◽

Ho-Jeong Lee ◽

...

Keyword(s):

Signaling Pathway ◽

Inflammatory Mediators ◽

Inflammatory Diseases ◽

Mapk Signaling ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Two Dimensional Gel Electrophoresis ◽

Necrosis Factor Alpha ◽

Mitogen Activated Protein ◽

Anti Inflammatory

Scutellaria baicalensisGeorgi has been used as traditional medicine for treating inflammatory diseases, hepatitis, tumors, and diarrhea in Asia. Hence, we investigated the anti-inflammatory effect and determined the molecular mechanism of action of flavonoids isolated from KoreanS. baicalensisG. in lipopolysaccharide- (LPS-) stimulated RAW 264.7 macrophages. A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was performed to examine cytotoxicity of the flavonoids at various concentrations of 10, 40, 70, and 100 µg/mL. No cytotoxicity was observed in RAW 264.7 cells at these concentrations. Furthermore, the flavonoids decreased production of inflammatory mediators such as inducible nitric oxide synthase, cyclooxygenase-2, interleukin-6, and tumor necrosis factor-alpha and inhibited phosphorylation of nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinases (MAPKs) in LPS-induced RAW 264.7 cells. Moreover, to identify the differentially expressed proteins in RAW 264.7 cells of the control, LPS-treated, and flavonoid-treated groups, two-dimensional gel electrophoresis and mass spectrometry were conducted. The identified proteins were involved in the inflammatory response and included PRKA anchor protein and heat shock protein 70 kD. These findings suggest that the flavonoids isolated fromS. baicalensisG. might have anti-inflammatory effects that regulate the expression of inflammatory mediators by inhibiting the NF-κB signaling pathway via the MAPK signaling pathway in RAW 264.7 cells.

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The Anti-Inflammatory Effects of Shinbaro3 Is Mediated by Downregulation of the TLR4 Signalling Pathway in LPS-Stimulated RAW 264.7 Macrophages

Mediators of Inflammation ◽

10.1155/2018/4514329 ◽

2018 ◽

Vol 2018 ◽

pp. 1-14 ◽

Cited By ~ 6

Author(s):

Hwa-Jin Chung ◽

Wonil Koh ◽

Won Kyung Kim ◽

Joon-Shik Shin ◽

Jinho Lee ◽

...

Keyword(s):

Protein Kinase ◽

Mitogen Activated Protein Kinase ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Dependent Manner ◽

Extracellular Signal ◽

Mitogen Activated Protein ◽

Raw 264.7 Macrophage Cells ◽

Anti Inflammatory ◽

Dose Dependent

Shinbaro3, a formulation derived from the hydrolysed roots of Harpagophytum procumbens var. sublobatum (Engl.) Stapf, has been clinically used in the pharamacopuncture treatment of arthritis in Korea. In the present study, Shinbaro3 inhibited NO generation in LPS-induced RAW 264.7 cells in a dose-dependent manner. Shinbaro3 also downregulated the mRNA and protein expression of inflammatory mediators in a dose-dependent manner. Three mechanisms explaining the effects of Shinbaro3 in RAW 264.7 cells were identified as follows: (1) inhibition of the extracellular signal-regulated kinase 1 and 2 (ERK1/2), stress-activated protein kinase (SAPK)/c-Jun N-terminal protein kinase (JNK), and p38 mitogen-activated protein kinase (MAPK) pathways; (2) suppression of IκB kinase-α/β (IKK-α/β) phosphorylation and nuclear factor-kappa B (NF-κB) subunits in the NF-κB pathway, which are involved in MyD88-dependent signalling; and (3) downregulation of IFN-β mRNA expression via inhibition of interferon regulatory factor 3 (IRF3) and Janus-activated kinase 1 (JAK1)/signal transducer and activator of transcription 1 (STAT1) phosphorylation, which is involved in TRIF-dependent signalling. Shinbaro3 exerted anti-inflammatory effects in LPS-stimulated RAW 264.7 macrophage cells through modulation of the TLR4/MyD88 pathways, suggesting that Shinbaro3 is a novel anti-inflammatory therapeutic candidate in the field of pharmacopuncture.

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Inhibitory Effects of Aucklandia lappa Decne. Extract on Inflammatory and Oxidative Responses in LPS-Treated Macrophages

Molecules ◽

10.3390/molecules25061336 ◽

2020 ◽

Vol 25 (6) ◽

pp. 1336

Author(s):

Jae Sung Lim ◽

Sung Ho Lee ◽

Sang Rok Lee ◽

Hyung-Ju Lim ◽

Yoon-Seok Roh ◽

...

Keyword(s):

Nitric Oxide ◽

Nuclear Translocation ◽

Radical Scavenging ◽

Gastric Ulcers ◽

Raw 264.7 Cells ◽

No Production ◽

Raw 264.7 ◽

Related Factor ◽

Proinflammatory Mediators ◽

Anti Inflammatory

Aucklandia lappa Decne., known as “Mok-hyang” in Korea, has been used for the alleviation of abdominal pain, vomiting, diarrhea, and stress gastric ulcers in traditional oriental medicine. We investigated the anti-inflammatory and antioxidative effects of the ethanol extract of Aucklandia lappa Decne. (ALDE) in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. ALDE significantly inhibited the LPS-induced nitric oxide (NO) production and reduced inducible nitric oxide synthase (iNOS) expression in RAW 264.7 cells. The production of other proinflammatory mediators, including COX-2, interleukin (IL)-6, IL-1β, and tumor necrosis factor (TNF)-α, was reduced by ALDE in LPS-stimulated RAW 264.7 cells. The mechanism underlying the anti-inflammatory effects of ALDE was elucidated to be the suppression of LPS-induced nuclear translocation of p65, followed by the degradation of IκB and the inhibition of the phosphorylation of mitogen-activated protein kinases (MAPK). In addition, ALDE showed enhanced radical scavenging activity. The antioxidant effect of ALDE was caused by the enhanced expression of heme oxygenase (HO-1) via stabilization of the expression of the nuclear transcription factor E2-related factor 2 (Nrf2) pathway. Collectively, these results indicated that ALDE not only exerts anti-inflammatory effects via the suppression of the NF-κB and MAPK pathways but also has an antioxidative effect through the activation of the Nrf2/HO-1 pathway.

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Analgesic and Antiinflammatory Activities of the Aqueous Extract fromPlectranthus amboinicus(Lour.) Spreng. BothIn VitroandIn Vivo

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2012/508137 ◽

2012 ◽

Vol 2012 ◽

pp. 1-11 ◽

Cited By ~ 19

Author(s):

Yung-Jia Chiu ◽

Tai-Hung Huang ◽

Chuan-Sung Chiu ◽

Tsung-Chun Lu ◽

Ya-Wen Chen ◽

...

Keyword(s):

Aqueous Extract ◽

Nuclear Translocation ◽

Folk Medicine ◽

Raw 264.7 Cells ◽

Necrosis Factor Alpha ◽

Proinflammatory Mediators ◽

Anti Inflammatory ◽

Plectranthus Amboinicus

Plectranthus amboinicus(Lour.) Spreng. is a native Labiatae plant of Taiwan. The plants are commonly used in Chinese folk medicine for the treatment of cough, fever, sore throats, mumps, and mosquito bite. The aim of this study was to investigate the analgesic and antiinflammatory properties of the aqueous extract fromPlectranthus amboinicus(PA)in vivoandin vitro. PA inhibited pain induced by acetic acid and formalin, and inflammation induced by carrageenan. The anti-inflammatory effect of PA was related to modulating antioxidant enzymes' activities in the liver and decreasing the Malondialdehyde (MDA) level and the production of tumor necrosis factor alpha (TNF-α), and cyclooxygenase2 (COX-2) in edema-paw tissue in mice.In vitrostudies show that PA inhibited the proinflammatory mediators in RAW 264.7 cells stimulated with lipopolysaccharide (LPS). PA blocked the degradation of IκB-αand nuclear translocation of NF-κB p65 subunit. Finally, the amount of carvacrol in the aqueous extract of PA was 1.88 mg/g extract. Our findings suggest that PA has analgesic and anti-inflammatory activities. These effects were mediated by inhibiting the proinflammatory mediators through blocking NF-κB activation. Meanwhile, the effects observed in this study provide evidence for folkloric uses ofPlectranthus amboinicus(Lour.) Spreng. in relieving pain and inflammation.

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Suppressive Effect on Lipopolysaccharide-Induced Proinflammatory Mediators byCitrus aurantium L.in Macrophage RAW 264.7 Cells via NF-κB Signal Pathway

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2011/248592 ◽

2011 ◽

Vol 2011 ◽

pp. 1-12 ◽

Cited By ~ 17

Author(s):

Sang-Rim Kang ◽

Dae-Yong Han ◽

Kwang-Il Park ◽

Hyeon-Soo Park ◽

Yong-Bae Cho ◽

...

Keyword(s):

Proinflammatory Cytokines ◽

Nuclear Translocation ◽

Suppressive Effect ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Dependent Manner ◽

Citrus Fruits ◽

Proinflammatory Mediators ◽

Cox 2 ◽

Anti Inflammatory

Citrus fruits have been used as an edible fruit and a traditional medicine since ancient times. In particular, the peels of immature citrus fruits are used widely in traditional herbal medicine in Korea, as they are believed to contain bioactive components exerting anti-inflammatory activity. This study examined whether the crude methanol extract ofCitrus aurantium L.(CME) has a suppressive effect on inducible enzymes and proinflammatory cytokines by inhibiting the NF-κB pathway in LPS-stimulated macrophage RAW 264.7 cells. The cells were pretreated with the indicated concentrations of CME (5, 10, 20, and 50 μg/mL) and then treated with LPS (1 μg/mL). The results showed that CME (10, 20, and 50 μg/mL) inhibited the LPS- (1 μg/mL) induced mRNA and protein expression of iNOS in macrophage Raw 264.7 cells. In addition, the expression of COX-2 was inhibited at the mRNA and protein levels by CME in a dose-dependent manner. The mRNA expression of proinflammatory cytokines, such as TNF-αand IL-6, were markedly reduced by CME (10, 20, and 50 μg/mL). Moreover, CME clearly suppressed the nuclear translocation of the NF-κB p65 subunits, which was correlated with its inhibitory effect on I-κB phosphorylation. These results suggest that CME has anti-inflammatory properties by modulating the expression of COX-2, iNOS, and proinflammatory cytokines, such as TNF-αand IL-6, in macrophage RAW 264.7 cells via the NF-κB pathway.

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Anti-Inflammatory and Antioxidant Effects of Soroseris hirsuta Extract by regulating iNOS/NF-κB and NRF2/HO-1 Pathways in Murine Macrophage RAW 264.7 Cells

Applied Sciences ◽

10.3390/app11104711 ◽

2021 ◽

Vol 11 (10) ◽

pp. 4711

Author(s):

Woo Jin Lee ◽

Wan Yi Li ◽

Sang Woo Lee ◽

Sung Keun Jung

Keyword(s):

Nitric Oxide ◽

Nuclear Translocation ◽

Antioxidant Properties ◽

Raw 264.7 Cells ◽

Heme Oxygenase 1 ◽

Raw 264.7 ◽

Related Factor ◽

Iκb Kinase ◽

Anti Inflammatory ◽

Antioxidant Effects

Until now, the physiological effects of Soroseris hirsuta were primarily unknown. Here we have evaluated the anti-inflammatory and antioxidant effects of Soroseris hirsuta extract (SHE) on lipopolysaccharide (LPS)-activated murine macrophages RAW 264.7 cells. SHE inhibited nitric oxide expression and inducible nitric oxide synthase expression in RAW 264.7 cells treated with LPS. Moreover, SHE suppressed LPS-induced phosphorylation of IκB kinase, inhibitor of kappa B, p65, p38, and c-JUN N-terminal kinase. Western blot and immunofluorescence analyses showed that SHE suppressed p65 nuclear translocation induced by LPS. Furthermore, SHE inhibited the reactive oxygen species in LPS-treated RAW 264.7 cells. SHE significantly increased heme oxygenase-1 expression and the nuclear translocation of nuclear factor erythroid 2-related factor 2. SHE suppressed LPS-induced interleukin-1β mRNA expression in RAW 264.7 cells. Thus, SHE is a promising nutraceutical as it displays anti-inflammatory and antioxidant properties.

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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.

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Suppression of Nitric Oxide Synthase by Thienodolin in Lipopolysaccharide-stimulated RAW 264.7 Murine Macrophage Cells

Natural Product Communications ◽

10.1177/1934578x1200700625 ◽

2012 ◽

Vol 7 (6) ◽

pp. 1934578X1200700 ◽

Cited By ~ 1

Author(s):

Eun-Jung Park ◽

John M. Pezzuto ◽

Kyoung Hwa Jang ◽

Sang-Jip Nam ◽

Sergio A. Bucarey ◽

...

Keyword(s):

Nitric Oxide ◽

Nitric Oxide Synthase ◽

Nuclear Translocation ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Dependent Manner ◽

Signal Transducers ◽

Protein Levels ◽

Mitogen Activated Protein ◽

Oxide Synthase

The measurement of nitric oxide in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells is used as a model for evaluating the anti-inflammatory or chemopreventive potential of substances. Thienodolin, isolated from a Streptomyces sp. derived from Chilean marine sediment, inhibited nitric oxide production in LPS-stimulated RAW 264.7 cells (IC50 = 17.2 ± 1.2 μM). At both the mRNA and protein levels, inducible nitric oxide synthase (iNOS) was suppressed in a dose-dependent manner. Mitogen-activated protein kinases (MAPKs), one major upstream signaling pathway involved in the transcription of iNOS, were not affected by treatment of thienodolin. However, the compound blocked the degradation of IκBα resulting in inhibition of NF-κB p65 nuclear translocation, and inhibited the phosphorylation of signal transducers and activators of transcription 1 (STAT1) at Tyr701. This study supports further exploration of thienodolin as a potential therapeutic agent with a unique mechanistic activity.

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A Combination Extract of Gardeniae Fructus and Perillae Folium Exerts Anti-Inflammatory Effects on LPS-Activated RAW 264.7 Mouse Macrophages via an ER Stress-Induced CHOP Pathway

Processes ◽

10.3390/pr9091632 ◽

2021 ◽

Vol 9 (9) ◽

pp. 1632

Author(s):

Wansu Park

Keyword(s):

Er Stress ◽

P38 Mapk ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Mrna Levels ◽

Mapk Phosphorylation ◽

Inflammatory Reactions ◽

Inflammatory Protein ◽

Mouse Macrophages ◽

Anti Inflammatory

The aim of this study is to investigate the effects of a combination extract of Gardeniae Fructus and Perillae Folium (GP) on inflammatory reactions in lipopolysaccharide (LPS)-activated mouse macrophages RAW 264.7 cells. Multiplex cytokine assay, Fluo-4 calcium assay, Flow cytometry assay for phospho-P38 MAPK, and quantitative PCR were carried out. GP significantly reduced LPS-induced productions of macrophage inflammatory protein (MIP)-1α and monokine induced by gamma interferon (MIG) and release of intracellular calcium in LPS-activated RAW 264.7 cells. GP also significantly inhibited P38 MAPK phosphorylation and mRNA levels of Chop, Camk2a, Stat1, Stat3, Jak2, Fas, Nos2, and Ptgs2 in LPS-activated RAW 264.7 cells. Taken together, this study represents that GP exerts anti-inflammatory effects on LPS-activated RAW 264.7 cells via ER stress-induced CHOP pathway.

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