Pro-inflammatory Cytokines and Nitric Oxide Inhibitory Constituents from Cassia occidentalis Roots

The anti-inflammatory and cytotoxic activity of thirty-six extracts of nine Indian medicinal plants were determined by measuring the inhibition of production of nitric oxide (NO), interleukin 1beta (IL-1β) and tumor necrosis factor alpha (TNF-α) in lipopolysaccharide (LPS) stimulated RAW 264.7 cells. Their cytotoxic activity against macrophages was determined by MTT assay. The ethyl acetate (EtOAc) extract of Cassia occidentalis L. (roots) (IC50= 21.3 to 43.1 μg/mL) and Mimosa pudica (whole plant) (IC50 = 31.7 to 47.2 μg/mL) and the dichloromethane (DCM) extract of Leucas cephalotes (whole plant) (IC50= 46.8 to 49.3 μg/mL) exhibited significant anti-inflammatory activity by in vitro inhibition of the production of TNF-α, IL-1β and NO in LPS stimulated RAW 264.7 cells. Furthermore, the five compounds isolated from the ethyl acetate extract of Cassia occidentalis roots were found to suppress LPS-induced IL-1β, TNF-α and NO production in a concentration-dependent manner in these cells at IC50 values ranging from 22.5 to 97.4 μM. Emodin and chrysophanol were also found active in inhibiting pro-inflammatory cytokines in vivo. These findings justify an ethnopharmacological use of C occidentalis roots as an effective herbal remedy for the treatment and prevention of inflammation and associated ailments.

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Enhanced immune response by vacuoles isolated from Saccharomyces cerevisiae in RAW 264.7 macrophages

Bioscience Reports ◽

10.1042/bsr20211158 ◽

2021 ◽

Vol 41 (9) ◽

Author(s):

Su-Min Lee ◽

Wooil Choi ◽

Woo-Ri Shin ◽

Yang-Hoon Kim ◽

Jiho Min

Keyword(s):

Nitric Oxide ◽

Saccharomyces Cerevisiae ◽

Immune Response ◽

Inflammatory Cytokines ◽

Membrane Vesicles ◽

Raw 264.7 Cells ◽

No Production ◽

Raw 264.7 ◽

Tnf Α ◽

Pro Inflammatory Cytokines

Abstract Vacuoles are membrane vesicles in eukaryotic cells, the digestive system of cells that break down substances absorbed outside the cell and digest the useless components of the cell itself. Researches on anticancer and intractable diseases using vacuoles are being actively conducted. The practical application of the present study to animals requires the determination of the biocompatibility of vacuole. In the present study, we evaluated the effects of vacuoles isolated from Saccharomyces cerevisiae in RAW 264.7 cells. This showed a significant increase in the production of nitric oxide (NO) produced by macrophage activity. Using Reactive Oxygen Species (ROS) assay, we identified that ROS is increased in a manner dependent on vacuole concentration. Western blot analysis showed that vacuole concentration-dependently increased protein levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2). Therefore, iNOS expression was stimulated to induce NO production. In addition, pro-inflammatory cytokines levels promoted, such as interleukin (IL) 6 (IL-6) and tumor necrosis factor (TNF) α (TNF-α). In summary, vacuoles activate the immune response of macrophages by promoting the production of immune-mediated transporters NO, ROS, and pro-inflammatory cytokines.

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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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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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miR-150 and SRPK1 regulate AKT3 expression to participate in LPS-induced inflammatory response

Innate Immunity ◽

10.1177/17534259211018800 ◽

2021 ◽

Vol 27 (4) ◽

pp. 343-350

Author(s):

Yanfen Yao ◽

Hong Wang ◽

Xueqin Xi ◽

Wei Sun ◽

Junke Ge ◽

...

Keyword(s):

Inflammatory Cytokines ◽

Inflammatory Responses ◽

Raw 264.7 Cells ◽

Control Group ◽

Raw 264.7 ◽

Sr Protein ◽

Over Expression ◽

Tnf Α ◽

Potential Factor ◽

Pro Inflammatory Cytokines

miR-150 was found to target the 3′-untranslated regions of AKT3, and the AKT pathway was affected by SR protein kinase 1 (SRPK1). However, the expression and significance of miR-150, AKT3 and SRPK1 in acute lung injury (ALI) were not clear. Here, we found that the expression of miR-150 was significantly reduced, while the expression of AKT3 and SRPK1 were markedly increased in LPS-treated A549, THP-1 and RAW 264.7 cells. miR-150 significantly decreased levels of pro-inflammatory cytokines IL-1β, IL-6 and TNF-α, reduced the expression of AKT3, but had no impact on SRPK1 expression compared with the control group in LPS-treated A549, THP-1 and RAW 264.7 cells. AKT3 silencing only reduced the production of pro-inflammatory cytokines and showed no effect on miR-150 and SRPK1 expression. Finally, we observed that miR-150 mimics and/or silencing of SRPK1 decreased the expression of AKT3 mRNA. Besides, over-expression of miR-150 or silencing of SRPK1 also reduced the expression of AKT3 protein, which exhibited the lowest level in the miR-150 mimics plus si-SRPK1 group. However, si-SRPK1 had no effect on miR-150 level. In conclusion, miR-150 and SRPK1 separately and cooperatively participate into inflammatory responses in ALI through regulating AKT3 pathway. Increased miR-150 and silenced SRPK1 may be a novel potential factor for preventing and treating more inflammatory lung diseases.

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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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Inhibitory Effect of Ulva fasciata and Desmarestia viridis on the Production of Nitric Oxide, Prostaglandin E2, and Pro-inflammatory Cytokines in RAW 264.7 Cells

Journal of Applied Pharmaceutical Science ◽

10.7324/japs.2013.3901 ◽

2013 ◽

Keyword(s):

Nitric Oxide ◽

Prostaglandin E2 ◽

Inflammatory Cytokines ◽

Inhibitory Effect ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Ulva Fasciata ◽

Pro Inflammatory Cytokines

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Two methoxy derivatives of resveratrol, 3,3′,4,5′-tetramethoxy-trans-stilbene and 3,4′,5-trimethoxy-trans-stilbene, suppress lipopolysaccharide-induced inflammation through inactivation of MAPK and NF-κB pathways in RAW 264.7 cells

Chinese Medicine ◽

10.1186/s13020-021-00480-9 ◽

2021 ◽

Vol 16 (1) ◽

Author(s):

Chunxiu Zhou ◽

Xutao Zhang ◽

Cheng-Chao Ruan ◽

Wai San Cheang

Keyword(s):

Nitric Oxide ◽

Signaling Pathways ◽

Inflammatory Cytokines ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Dependent Manner ◽

Trans Stilbene ◽

No Release ◽

Derivatives Of ◽

Pro Inflammatory Cytokines

Abstract Background 3,3′,4,5′-tetramethoxy-trans-stilbene (3,3′,4,5′-TMS) and 3,4′,5-trimethoxy-trans-stilbene (3,4′,5-TMS) are two methoxy derivatives of resveratrol. Previous researches have proved that resveratrol and its analogues have anti-inflammatory effect through suppressing mitogen-activated protein kinase (MAPK) and nuclear factor-κB (NF-κB) signaling pathways. This study aims to study whether 3,3′,4,5′-TMS and 3,4′,5-TMS alleviate inflammation and the underlying mechanism. Methods RAW 264.7 macrophage cells were treated with lipopolysaccharide (LPS) to induce inflammation and pretreated with 3,3′,4,5′-TMS or 3,4′,5-TMS. Cell viability was measured with the 3-(4,5)-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Nitric oxide (NO) release was detected by Griess reagent. The secretions of pro-inflammatory cytokines were assessed by ELISA kits. Protein expressions of signaling molecules were determined by Western blotting. Reactive oxygen species (ROS) production was detected by fluorescence staining and malondialdehyde (MDA) assay. Results 3,3′,4,5′-TMS and 3,4′,5-TMS suppressed LPS-induced NO release and pro-inflammatory cytokines (IL-6 and TNF-α) secretions in a dose-dependent manner in RAW 264.7 cells. 3,3′,4,5′-TMS and 3,4′,5-TMS significantly down-regulated the LPS-induced expressions of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS), and partially suppressed the activation of MAPK (phosphorylation of p38, JNK, ERK), and NF-κB (phosphorylation of IKKα/β, p65 and IκBα) signaling pathways; where phosphorylation of ERK and p65 was mildly but not significantly decreased by 3,3′,4,5′-TMS. LPS-induced NF-κB/p65 nuclear translocation was inhibited by both 3,3′,4,5′-TMS and 3,4′,5-TMS. Moreover, both resveratrol derivatives decreased the ROS levels. Conclusions 3,3′,4,5′-TMS and 3,4′,5-TMS significantly suppress LPS-induced inflammation in RAW 264.7 cells through inhibition of MAPK and NF-κB signaling pathways and also provide anti-oxidative effect. This study reveals potential therapeutic applications of 3,3′,4,5′-TMS and 3,4′,5-TMS for inflammatory diseases.

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Comparing the protective effects of resveratrol, curcumin and sulforaphane against LPS/IFN-γ-mediated inflammation in doxorubicin-treated macrophages

Scientific Reports ◽

10.1038/s41598-020-80804-1 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Haidy A. Saleh ◽

Eman Ramdan ◽

Mohey M. Elmazar ◽

Hassan M. E. Azzazy ◽

Anwar Abdelnaser

Keyword(s):

Nitric Oxide ◽

Inflammatory Response ◽

Raw 264.7 Cells ◽

Inos Mrna ◽

No Production ◽

Raw 264.7 ◽

Mrna Levels ◽

Protective Effects ◽

Tnf Α ◽

Ifn Γ

AbstractDoxorubicin (DOX) chemotherapy is associated with the release of inflammatory cytokines from macrophages. This has been suggested to be, in part, due to DOX-mediated leakage of endotoxins from gut microflora, which activate Toll-like receptor 4 (TLR4) signaling in macrophages, causing severe inflammation. However, the direct function of DOX on macrophages is still unknown. In the present study, we tested the hypothesis that DOX alone is incapable of stimulating inflammatory response in macrophages. Then, we compared the anti-inflammatory effects of curcumin (CUR), resveratrol (RES) and sulforaphane (SFN) against lipopolysaccharide/interferon-gamma (LPS/IFN-γ)-mediated inflammation in the absence or presence of DOX. For this purpose, RAW 264.7 cells were stimulated with LPS/IFN-γ (10 ng/mL/10 U/mL) in the absence or presence of DOX (0.1 µM). Our results showed that DOX alone is incapable of stimulating an inflammatory response in RAW 264.7 macrophages. Furthermore, after 24 h of incubation with LPS/IFN-γ, a significant increase in tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and inducible nitric oxide synthase (iNOS) mRNA levels was observed. Similarly, nitric oxide (NO) production and TNF-α and IL-6 protein levels were significantly upregulated. Moreover, in LPS/IFN-γ-treated macrophages, the microRNAs (miRNAs) miR-146a, miR-155, and miR-21 were significantly overexpressed. Interestingly, upon testing CUR, RES, and SFN against LPS/IFN-γ-mediated inflammation, only SFN was able to significantly reverse the LPS/IFN-γ-mediated induction of iNOS, TNF-α and IL-6 and attenuate miR-146a and miR-155 levels. In conclusion, SFN, at the transcriptional and posttranscriptional levels, exhibits potent immunomodulatory action against LPS/IFN-γ-stimulated macrophages, which may indicate SFN as a potential treatment for DOX-associated inflammation.

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A New Flavonol From Saussurea involucrata With Anti-Inflammatory Activity

Natural Product Communications ◽

10.1177/1934578x211007638 ◽

2021 ◽

Vol 16 (5) ◽

pp. 1934578X2110076

Author(s):

Sheng Pan ◽

Zi-Guan Zhu

Keyword(s):

Nitric Oxide ◽

Aerial Part ◽

Raw 264.7 Cells ◽

Raw 264.7 ◽

Spectroscopic Methods ◽

Soluble Extract ◽

Saussurea Involucrata ◽

Tnf Α ◽

Anti Inflammatory

A new flavonol named 6-(2'',3''-epoxy-3''-methylbutyl)-resokaempferol (1), together with five known compounds (2-6) were isolated from the EtOAc-soluble extract of the aerial part of Saussurea involucrata. Their structures were elucidated on the basis of spectroscopic methods. All compounds were evaluated for their anti-inflammatory effects by measuring the production of nitric oxide (NO) and TNF-α in vitro. Among them, compound 1 showed potential inhibitory activity on the production of NO and TNF-α in LPS-induced RAW 264.7 cells with IC50 values of 48.0 ± 1.5 and 41.4 ± 1.7 µM, respectively.

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