Homocysteine elicits an M1 phenotype in murine macrophages through an EMMPRIN-mediated pathway

2015 ◽  
Vol 93 (7) ◽  
pp. 577-584 ◽  
Author(s):  
Lee J. Winchester ◽  
Sudhakar Veeranki ◽  
Srikanth Givvimani ◽  
Suresh C. Tyagi
Keyword(s):  
Nitric Oxide ◽  
Matrix Metalloproteinase ◽  
Cell Lines ◽  
Inflammatory Diseases ◽  
Raw 264.7 ◽  
Macrophage Phenotype ◽  
Raw 264.7 Macrophages ◽  
M1 Macrophage ◽  
Endothelial Nitric Oxide ◽  
The Impact

Introduction: Hyperhomocysteinemia (HHcy) is associated with inflammatory diseases and is known to increase the production of reactive oxygen species (ROS), matrix metalloproteinase (MMP)-9, and inducible nitric oxide synthase, and to decrease endothelial nitric oxide production. However, the impact of HHcy on macrophage phenotype differentiation is not well-established. It has been documented that macrophages have 2 distinct phenotypes: the “classically activated/destructive” (M1), and the “alternatively activated/constructive” (M2) subtypes. We hypothesize that HHcy increases M1 macrophage differentiation through extracellular matrix metalloproteinase inducer (EMMPRIN), a known inducer of matrix metalloproteinases. Methods: murine J774A.1 and Raw 264.7 macrophages were treated with 100 and 500 μmol/L Hcy, respectively, for 24 h. Samples were analyzed using Western blotting and immunocytochemistry. Results: Homocysteine treatment increased cluster of differentiation 40 (CD40; M1 marker) in J774A.1 and Raw 264.7 macrophages. MMP-9 was induced in both cell lines. EMMPRIN protein expression was also increased in both cell lines. Blocking EMMPRIN function by pre-treating cells with anti-EMMPRIN antibody, with or without Hcy, resulted in significantly lower expression of CD40 in both cell lines by comparison with the controls. A DCFDA assay demonstrated increased ROS production in both cell lines with Hcy treatment when compared with the controls. Conclusion: Our results suggest that HHcy results in an increase of the M1 macrophage phenotype. This effect seems to be at least partially mediated by EMMPRIN induction.

scholarly journals Inhibition of nitric oxide production in lipopolysaccharide-activated RAW 264.7 macrophages by Jeju plant extracts

2009 ◽  
Vol 2 (4) ◽  
pp. 245-249 ◽  
Author(s):  
Eun-Jin Yang ◽  
Eun-Young Yim ◽  
Gwanpil Song ◽  
Gi-Ok Kim ◽  
Chang-Gu Hyun
Keyword(s):  
Nitric Oxide ◽  
Plant Extracts ◽  
Inhibitory Activity ◽  
Inflammatory Diseases ◽  
No Production ◽  
Raw 264.7 ◽  
Nitric Oxide Production ◽  
Raw 264.7 Macrophages ◽  
Oxide Production ◽  
Potent Inhibition

Inhibition of nitric oxide production in lipopolysaccharide-activated RAW 264.7 macrophages by Jeju plant extractsNitric oxide (NO) produced in large amounts by inducible nitric oxide synthase (iNOS) is known to be responsible for the vasodilation and hypotension observed during septic shock and inflammation. Thus, inhibitors of iNOS may be useful candidates for the treatment of inflammatory diseases accompanied by the overproduction of NO. In this study, we prepared alcoholic extracts of Jeju plants and screened them for their inhibitory activity against NO production in lipopolysaccharide (LPS)-activated macrophages. Among the 260 kinds of plant extract tested, 122 extracts showed potent inhibitory activity towards NO production by more than 25% at a concentration of 100 μg/mL. Plants such asMalus sieboldii, Vaccinium oldhamii, Corylus hallaisanensis, Carpinus laxiflora, Styrax obassia, andSecurinega suffruticosashowed the most potent inhibition (above 70%) at a concentration of 100 μg/mL. The cytotoxic effects of the plant extracts were determined by colorimetric MTT assays and most plant extracts exhibited only moderate cytotoxicity at 100 μg/mL. Therefore, these plants should be considered promising candidates for the further purification of bioactive compounds and would be useful for the treatment of inflammatory diseases accompanying overproduction of NO.

scholarly journals Neolitsea Sericea Essential Oil Attenuates LPS-induced Inflammation in RAW 264.7 Macrophages by Suppressing NF-κB and MAPK Activation

2010 ◽  
Vol 5 (8) ◽  
pp. 1934578X1000500 ◽  
Author(s):  
Weon-Jong Yoon ◽  
Ji-Young Moon ◽  
Ji-Yong Kang ◽  
Gi-Ok Kim ◽  
Nam Ho Lee ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Essential Oil ◽  
Inflammatory Diseases ◽  
Chemical Constituents ◽  
Nuclear Factor Κb ◽  
Prostaglandin E ◽  
Raw 264.7 ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Raw 264.7 Macrophages

The chemical composition and antiinflammatory activities of hydrodistilled essential oil from Neolitsea sericea leaves (NSE) have been investigated for the first time. The chemical constituents of NSE were analysed by GC-MS and found to include sericenine (32.3%), sabinene (21.0%), trans-β-ocimene (13.3%), β-caryophyllene (4.8%), and 4-terpineol (4.2%). The effects of NSE on nitric oxide (NO), prostaglandin E2 (PGE2), tumour necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 production in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages were also examined. Pro-inflammatory cytokine and mediator tests indicated that NSE has excellent dose-dependent inhibitory activities. To further examine the mechanism responsible for the inhibition of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression by NSE, we examined the effect of NSE on nuclear factor-κB (NF-κB) activation and the phosphorylation of mitogen-activated protein kinases (MAPK). NSE inhibited NF-κB activation by LPS, and this was associated with the abrogation of IκB-α phosphorylation and subsequent decreases in nuclear p50 and p65 protein levels. Further, the phosphorylation of p38, ERK and JNK was suppressed by NSE in a concentration-dependent manner. These results suggest that NSE exerts antiinflammatory effects in LPS-stimulated RAW 264.7 macrophages by inhibition of NF-κB activation and MAPK phosphorylation, and, therefore, may be useful for treatment of inflammatory diseases.

scholarly journals In-vitro Anti-Cancer and Anti-Inflammatory Screening of Dodonaea viscosa

2021 ◽  
pp. 186-192
Author(s):  
R. Ramkumar ◽  
S. K. Periyasamy ◽  
B. R. Venkatraman ◽  
K. G. Sekar
Keyword(s):  
Cell Lines ◽  
Raw 264.7 ◽  
Dodonaea Viscosa ◽  
Raw 264.7 Macrophages ◽  
Tnf Α ◽  
Cervical Disease ◽  
Hct 116 ◽  
The Impact ◽  
Mcf 7

Background: The current investigation was done to assess the in vitro anticancer property of Dodonaea viscosa (D. viscosa) in three malignant growth cell lines and mitigating impact in RAW 264.7 macrophages. Methods: The hydroalcoholic remove D. viscosa was ready and tried against HCT-116 colon malignancy, MCF-7 bosom disease HeLa cervical disease cell lines. The cytotoxicity of concentrate was affirmed by MTT cheeky. The calming movement of concentrate was assessed utilizing LPS invigorated RAW 264.7 macrophages and the degree of incendiary middle people was estimated. Results: The anticancer impact of D. viscosa onHCT-116, MCF-7 and HeLa cell line with the IC50 worth of 60.43 ± 0.76 μg/ml,75.26 ± 0.45 μg/ml and 72.12 ± 0.87μg/ml individually. Further, in LPS stimulatedRAW264.7 macrophage cells, treatment with D. viscosa extract altogether decreased the raised level of NO, TNF-α and PGE2. Conclusion: This examination gave the proof to D. viscosa an anticancer and mitigating specialist. Further bioactive confinement and atomic examinations are needed to prove the impact of plant remove.

scholarly journals Targeting MAPK/NF-κB Pathways in Anti-Inflammatory Potential of Rutaecarpine: Impact on Src/FAK-Mediated Macrophage Migration

2021 ◽  
Vol 23 (1) ◽  
pp. 92
Author(s):  
Thanasekaran Jayakumar ◽  
Kao-Chang Lin ◽  
Chao-Chien Chang ◽  
Chih-Wei Hsia ◽  
Manjunath Manubolu ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Molecular Mechanisms ◽  
Nuclear Translocation ◽  
Inflammatory Diseases ◽  
Biological Activities ◽  
Inhibitory Effect ◽  
Vital Role ◽  
Raw 264.7 ◽  
Raw 264.7 Macrophages ◽  
Anti Inflammatory

Studies have discovered that different extracts of Evodia rutaecarpa and its phytochemicals show a variety of biological activities associated with inflammation. Although rutaecarpine, an alkaloid isolated from the unripe fruit of E. rutaecarpa, has been exposed to have anti-inflammatory properties, the mechanism of action has not been well studied. Thus, this study investigated the molecular mechanisms of rutaecarpine (RUT) in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. RUT reserved the production of nitric oxide (NO) and the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis factor (TNF-α), and interleukin (IL)-1β in the LPS-induced macrophages. RUT showed an inhibitory effect on the mitogen-activated protein kinases (MAPKs), and it also inhibited nuclear transcription factor kappa-B (NF-κB) by hindering IκBα and NF-κB p65 phosphorylation and p65 nuclear translocation. The phospho-PI3K and Akt was concentration-dependently suppressed by RUT. However, RUT not only suggestively reduced the migratory ability of macrophages and their numbers induced by LPS but also inhibited the phospho-Src, and FAK. Taken together, these results indicate that RUT participates a vital role in the inhibition of LPS-induced inflammatory processes in RAW 264.7 macrophages and that the mechanisms involve PI3K/Akt and MAPK-mediated downregulation of NF-κB signaling pathways. Notably, reducing the migration and number of cells induced by LPS via inhibiting of Src/FAK pathway was also included to the anti-inflammatory mechanism of RUT. Therefore, RUT may have potential benefits as a therapeutic agent against chronic inflammatory diseases.

scholarly journals Antioxidant and Anti-Inflammatory Effects of Herbal Formula SC-E3 in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages

2017 ◽  
Vol 2017 ◽  
pp. 1-13 ◽  
Author(s):  
Soo Chil Lee ◽  
Young-Won Kwon ◽  
Ju-Yeon Park ◽  
Sung Yun Park ◽  
Ju-Hee Lee ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Free Radical ◽  
Inflammatory Diseases ◽  
Heme Oxygenase 1 ◽  
Prostaglandin E ◽  
Raw 264.7 ◽  
Herbal Formula ◽  
Raw 264.7 Macrophages ◽  
Wide Range ◽  
Anti Inflammatory

SC-E3 is a novel herbal formula composed of five oriental medicinal herbs that are used to treat a wide range of inflammatory diseases in Korean traditional medicine. In this study, we sought to determine the effects of SC-E3 on free radical generation and inflammatory response in lipopolysaccharide- (LPS-) treated RAW 264.7 macrophages and the molecular mechanism involved. The ethanol extract of SC-E3 showed good free radical scavenging activity and inhibited LPS-induced reactive oxygen species generation. SC-E3 significantly inhibited the production of the LPS-induced inflammatory mediators, nitric oxide and prostaglandin E2, by suppressing the expressions of inducible nitric oxide synthase and cyclooxygenase-2, respectively. SC-E3 also prevented the secretion of the proinflammatory cytokines, IL-1β, TNF-α, and IL-6, and inhibited LPS-induced NF-κB activation and the mitogen-activated protein kinase (MAPK) pathway. Furthermore, SC-E3 induced the expression of heme oxygenase-1 (HO-1) by promoting the nuclear translocation and transactivation of Nrf2. Taken together, these results suggest that SC-E3 has potent antioxidant and anti-inflammatory effects and that these effects are due to the inhibitions of NF-κB and MAPK and the induction of Nrf2-mediated HO-1 expression in macrophages. These findings provide scientific evidence supporting the potential use of SC-E3 for the treatment and prevention of various inflammatory diseases.

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

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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