scholarly journals Bioactive Phytochemicals from Mulberry: Potential Anti-Inflammatory Effects in Lipopolysaccharide-Stimulated RAW 264.7 Macrophages

2021 ◽  
Vol 22 (15) ◽  
pp. 8120
Author(s):  
Dahae Lee ◽  
Seoung Rak Lee ◽  
Ki Sung Kang ◽  
Ki Hyun Kim
Keyword(s):  
Nitric Oxide ◽  
Ethanol Extract ◽  
Phytochemical Analysis ◽  
Raw 264.7 ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Raw 264.7 Macrophages ◽  
Iκb Kinase ◽  
Cox 2 ◽  
Anti Inflammatory

The fruits of the mulberry tree (Morus alba L.), known as white mulberry, have been consumed in various forms, including tea, beverages, and desserts, worldwide. As part of an ongoing study to discover bioactive compounds from M. alba fruits, the anti-inflammatory effect of compounds from M. alba were evaluated in lipopolysaccharide (LPS)-stimulated mouse RAW 264.7 macrophages. Phytochemical analysis of the ethanol extract of the M. alba fruits led to the isolation of 22 compounds. Among the isolated compounds, to the best of our knowledge, compounds 1, 3, 5, 7, 11, 12, and 14–22 were identified from M. alba fruits for the first time in this study. Inhibitory effects of 22 compounds on the production of the nitric oxide (NO) known as a proinflammatory mediator in LPS-stimulated RAW 264.7 macrophages were evaluated using NO assays. Western blot analysis was performed to evaluate the anti-inflammatory effects of cyclo(L-Pro-L-Val) (5). We evaluated whether the anti-inflammatory effects of cyclo(L-Pro-L-Val) (5) following LPS stimulation in RAW 264.7 macrophages occurred because of phosphorylation of IκB kinase alpha (IKKα), IκB kinase beta (IKKβ), inhibitor of kappa B alpha (IκBα), nuclear factor kappa B (NF-κB) and activations of inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2). Cyclo(L-Pro-L-Val) (5) significantly suppressed phosphorylations of IKKα, IKKβ, IκBα, and NF-κB and activations of iNOS and COX-2 in a concentration-dependent manner. Taken together, these results indicate that cyclo(L-Pro-L-Val) (5) can be considered a potential therapeutic agent for the treatment of inflammation-associated disorders.

scholarly journals Anti-inflammatory Effect of d-(+)-Cycloserine Through Inhibition of NF-κB and MAPK Signaling Pathways in LPS-Induced RAW 264.7 Macrophages

2020 ◽  
Vol 15 (4) ◽  
pp. 1934578X2092048 ◽  
Author(s):  
Hyun-Kyu Kang ◽  
Chang-Gu Hyun
Keyword(s):  
Nitric Oxide ◽  
Mtt Assay ◽  
Inflammatory Responses ◽  
Mapk Signaling ◽  
Prostaglandin E ◽  
Raw 264.7 ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Raw 264.7 Macrophages ◽  
Anti Inflammatory

Recently, additional therapeutic potentials of classical antibiotics are gaining considerable attention. The discovery of penicillin in the 1920s had a major impact on the history of human health. Penicillin has been used for the treatment for fatal microbial infections in humans and has led to the discovery of several new antibiotics. d-(+)-Cycloserine (DCS) is an antibiotic isolated from Streptomyces orchidaceous and is used in conjunction with other drugs in the treatment of tuberculosis. However, there have been no studies on the anti-inflammatory effects of DCS in RAW 264.7 macrophage cell line. To investigate the anti-inflammatory effects of DCS, we examined the ability of DCS to inhibit the inflammatory responses in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages in this study. Cell viability was analyzed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The cells were pretreated with various concentrations (2, 4, and 6 mM) of DCS, then treated with 1 μg/mL LPS to detect its anti-inflammatory effects. d-(+)-Cycloserine inhibited the production of nitric oxide (NO) in a concentration-dependent manner, and to some extent, inhibited the production of prostaglandin E2. Consistent with these findings, DCS suppressed the expression of pro-inflammatory cytokines such as interleukin (IL)-1β and IL-6. However, it had no effect on the expression of tumor necrosis factor-α. Western blot analysis demonstrated that DCS inhibited inducible nitric oxide synthase and suppressed cyclooxygenase type-2 (COX-2) expression. In addition, investigation of its effects on nuclear factor kappa B signaling showed that DCS inhibited phosphorylation of inhibitory kappa B-α (IκB-α) and increased intracellular IκB-α in a concentration-dependent manner. Furthermore, DCS inhibited the phosphorylation of LPS-induced extracellular signal-regulated kinase, however it did not affect phosphorylation of c-jun N-terminal kinase and p38. Further studies confirmed that the inhibition of phosphorylation of IκB-α was mediated through the inhibition of phosphoinositide 3-kinase/Akt (PI3K/Akt) pathway. To determine the applicability of DCS to the skin, cytotoxicity on HaCaT keratinocytes was measured following treatment with various concentrations (2, 4, 6, 8, and 10 mM) of DCS using MTT assay. These results suggest that DCS may be used as a potential drug for the treatment of inflammatory diseases.

scholarly journals Antioxidant activity of Rosa multiflora Thunb. flower extract and suppressive activity on proinflammatory mediator production in lipopolysaccharide- stimulated RAW 264.7 macrophages

2016 ◽  
Vol 6 (5) ◽  
pp. 265 ◽  
Author(s):  
Chungshil Kwak ◽  
Hye In Choi ◽  
Jiwon Yang
Keyword(s):  
Nitric Oxide ◽  
Antioxidant Activity ◽  
Ethanol Extract ◽  
Total Phenolic ◽  
Raw 264.7 ◽  
Rosa Multiflora ◽  
Raw 264.7 Macrophages ◽  
Tnf Α ◽  
Cox 2 ◽  
Anti Inflammatory

Background: Oxidative stress and inflammation are associated with various ageing-related chronic diseases. The fruits and roots of Rosa multiflora Thunb. have been used in medicine for the treatment of edema and inflammatory diseases in Eastern Asia. Dried Rosa multiflora Thunb. flower (RMF) have been consumed as a tea in Korea, but reports on the biological activity of RMF are lacking. We evaluated the in vitro antioxidant and anti-inflammatory effects of an ethanol extract from RMF as well as various solvent fractions from the extract.Methods: The ethanol extract (Et) of RMP was fractionated sequentially by hexane (Hx), dichloromethane (DM), ethylacetate (EA), n-butanol (Bt) and distilled water (DW). Total phenolic content and total flavonoid content, scavenging activities of the 2,2-diphenyl-1 picrylhydrazyl radical and 2,2′-azinobis-3-ethylbenzothiazoline-6-sulfonic acid radical and ferric-reducing antioxidant power were measured. Anti-inflammatory effects in terms of levels of nitric oxide and prostaglandin (PG) E2 and production of proinflammatory cytokines such as interleukin (IL)-6, tumor necrosis factor (TNF)-α in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages were measured, and expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 were measured by Western blot analysis.Results: EA treatment showed the highest total phenolic and total flavonoid content and strongest antioxidant activity, followed by Bt and Et, measured by three different methods. Treatment with Et and all fractions significantly suppressed (p<0.05) nitric oxide production in a dose-dependent manner in LPS-treated RAW 264.7 macrophages via reduction of expression of iNOS protein. Treatment with Et, DM and EA significantly suppressed (p<0.05) PGE2 production induced by LPS treatment, however, only Bt treatment significantly reduced (p<0.05) the expression of COX-2 protein. Treatment with DM, EA and Bt suppressed IL-6 production significantly (p<0.05) in LPS-treated RAW 264.7 macrophages, and treatment with Et, DM, EA and Bt suppressed TNF-α production significantly (p<0.05).Conclusions: These data suggest that the ethanol extract of Rosa multiflora Thunb. flower and its dichloromethan, ethylacetate and n-butanol fractions have potent antioxidant and/or anti-inflammatory activities.Keywords: Rosa multiflora Thunb. flower, antioxidant activity, anti-inflammatory activity, RAW 264 7 macrophages, cytokines, iNOS, COX-2 

scholarly journals (3β,16α)-3,16-Dihydroxypregn-5-en-20-one from the Twigs of Euonymus alatus (Thunb.) Sieb. Exerts Anti-Inflammatory Effects in LPS-Stimulated RAW-264.7 Macrophages

Molecules ◽  
2019 ◽  
Vol 24 (21) ◽  
pp. 3848 ◽  
Author(s):  
Seulah Lee ◽  
Dahae Lee ◽  
Su Cheol Baek ◽  
Mun Seok Jo ◽  
Ki Sung Kang ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Medicinal Plant ◽  
Phytochemical Analysis ◽  
No Production ◽  
Raw 264.7 ◽  
Raw 264.7 Macrophages ◽  
Euonymus Alatus ◽  
Cox 2 ◽  
Anti Inflammatory ◽  
Pharmacologically Active

To discover new pharmacologically active natural products, here, we performed the phytochemical analysis of a Korean medicinal plant. Euonymus alatus (Thunb.) Sieb. is a traditional medicinal plant that has been used as a remedy for various diseases in Asian countries. In particular, the cork cambium on the twigs of E. alatus has been used to treat dysmenorrhea, tumors, diabetes, and wound. Phytochemical analysis of the methanolic extract of E. alatus twigs led to the isolation of a sterol, which was identified as (3β,16α)-3,16-dihydroxypregn-5-en-20-one (1) by 1D and 2D nuclear magnetic resonance (NMR) spectroscopy and high-resolution electrospray ionization mass spectrometry. The stereochemistry of 1 was established with nuclear Overhauser effect spectroscopy (NOESY) analysis and comparison of electronic circular dichroism (ECD) data. To the best of our knowledge, the isolation of compound 1 from nature is first reported here, as well as the complete and revised NMR data assignment of 1. In lipopolysaccharide (LPS)-stimulated RAW-264.7 macrophages, compound 1 significantly inhibited nitric oxide (NO) production at an IC50 value of 12.54 ± 0.05 μM as well as the protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Moreover, the pre-treatment with compound 1 attenuated the LPS-induced phosphorylation of nuclear factor kappa B (NF-κB) p65 through the inhibition of the phosphorylation of IκB kinase alpha (IKKα), IKKβ, and inhibitor of kappa B alpha (IκBα). Compound 1 also inhibited the LPS-induced phosphorylation of p38, c-Jun NH2-terminal kinase (JNK), and extracellular signal-regulated kinase (ERK). Taken together, compound 1 may serve as an anti-inflammatory constituent of E. alatus twigs and its anti-inflammatory property is thought to be associated with the inhibition of NO production via suppression of iNOS and COX-2 expression through inhibition of IKKα/β, I-κBα and NF-κB p65 activation and downregulation of p38, JNK, and ERK mitogen-activated protein kinase signal pathways in RAW 264.7 macrophages. These findings also provide experimental evidence that compound 1 identified from E. alatus twigs could be a candidate for an anti-inflammatory agent.

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.

scholarly journals Oleifolioside A, a New Active Compound, Attenuates LPS-Stimulated iNOS and COX-2 Expression through the Downregulation of NF-κB and MAPK Activities in RAW 264.7 Macrophages

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Hai Yang Yu ◽  
Kyoung-Sook Kim ◽  
Young-Choon Lee ◽  
Hyung-In Moon ◽  
Jai-Heon Lee
Keyword(s):  
Nitric Oxide ◽  
Mapk Signaling ◽  
Binding Activity ◽  
Prostaglandin E ◽  
Raw 264.7 ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Raw 264.7 Macrophages ◽  
Dendropanax Morbifera ◽  
Cox 2

Oleifolioside A, a new triterpenoid compound isolated fromDendropanax morbiferaLeveille (D. morbifera), was shown in this study to have potent inhibitory effects on lipopolysaccharide (LPS-)stimulated nitric oxide (NO) and prostaglandin E2(PGE2) production in RAW 264.7 macrophages. Consistent with these findings, oleifolioside A was further shown to suppress the expression of LPS-stimulated inducible nitric oxide synthase (iNOS) and cyclooxigenase-2 (COX-2) in a dose-dependent manner at both the protein and mRNA levels and to significantly inhibit the DNA-binding activity and transcriptional activity of NF-κB in response to LPS. These results were found to be associated with the inhibition of the degradation and phosphorylation of IκB-αand subsequent translocation of the NF-κB p65 subunit to the nucleus. Inhibition of NF-κB activation by oleifolioside A was also shown to be mediated through the prevention of p38 MAPK and ERK1/2 phosphorylation. Taken together, our results suggest that oleifolioside A has the potential to be a novel anti-inflammatory agent capable of targeting both the NF-κB and MAPK signaling pathways.

Anti-Oxidative and Anti-Inflammatory Activity of Kenya Grade AA Green Coffee Bean Extracts

2020 ◽  
Author(s):  
In-Chul LEE ◽  
Jae-Sook LEE ◽  
Jeong-Hyun LEE ◽  
Yeona KIM ◽  
Wi-Young SO
Keyword(s):  
Nitric Oxide ◽  
Coffee Bean ◽  
Dependent Manner ◽  
Green Coffee ◽  
Cosmetic Products ◽  
Concentration Dependent Manner ◽  
Total Polyphenol ◽  
Cox 2 ◽  
Anti Inflammatory ◽  
Green Coffee Bean

Background: Kenya AA green coffee bean extracts were tested for natural ingredients used for anti-oxidative and anti-inflammatory purposes in cosmetic products Methods: Anti-oxidative activities were measured by total polyphenol, 1,1-diphenyl-2-picrylhydrazyl (DPPH), and the 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays. Anti-inflammatory activities were evaluated via nitric oxide (NO) assays, and through quantification of inducible nitric oxide synthase (iNOS), and cyclooxygenase-2 (COX-2) protein expression by western blotting. Data analyses were performed using independent Student’s t-tests, with statistical significance set at P < 0.05. Results: Total polyphenol content of water and ethanol extract was 169.0 ± 3.1 mg and 300.34 ± 16.6 mg tannic acid/g dry weight, respectively. The DPPH and ABTS radical scavenging activities of all the extracts were significantly increased in a concentration-dependent manner. Kenya AA green coffee bean extracts were toxic at a concentration of 1,000 µg/mL in RAW 264.7 cells. Anti-inflammatory activity as determined by NO assay showed that lipopolysaccharide (LPS)-induced NO was significantly inhibited following treatment with Kenya AA green coffee bean extracts in a concentration-dependent manner. iNOS and COX-2 protein expression was also significantly inhibited following treatment. Conclusion: These results highlight the potential of Kenya AA green coffee bean extracts as a naturally active anti-inflammatory agent in cosmetic products.

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

Molecules ◽  
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.

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 New 12,23-Epoxydammarane Type Saponins Obtained from Panax notoginseng Leaves and Their Anti-Inflammatory Activity

Molecules ◽  
2020 ◽  
Vol 25 (17) ◽  
pp. 3784
Author(s):  
Jingya Ruan ◽  
Ying Zhang ◽  
Wei Zhao ◽  
Fan Sun ◽  
Lifeng Han ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Panax Notoginseng ◽  
Inflammatory Activity ◽  
Raw 264.7 Cells ◽  
Raw 264.7 ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Necrosis Factor Alpha ◽  
Anti Inflammatory

Two new 12,23-epoxydammarane-type saponins, notoginsenosides NL-I (1) and NL-J (2), were isolated and identified from Panax notoginseng leaves through the combination of various chromatographies and extensive spectroscopic methods, as well as chemical reactions. Among them, notoginsenoside NL-J (2) had a new skeleton. Furthermore, the lipopolysaccharide (LPS)-induced RAW 264.7 macrophage model was used to identify the in vitro anti-inflammatory activity and mechanisms of compounds 1 and 2. Both of them exerted strong inhibition on nitric oxide (NO) productions in a concentration-dependent manner at 1, 10, and 25 μM. Moreover, these two compounds significantly decreased the secretion of tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL-6), cyclooxygenase-2 (COX-2), nuclear factor kappa-B (NF-κB/p65), and nitric-oxide synthase (iNOS) in LPS-activated RAW 264.7 cells.

scholarly journals Anti-Inflammatory Activity and ROS Regulation Effect of Sinapaldehyde in LPS-Stimulated RAW 264.7 Macrophages

Molecules ◽  
2020 ◽  
Vol 25 (18) ◽  
pp. 4089
Author(s):  
Seung-Hwa Baek ◽  
Tamina Park ◽  
Myung-Gyun Kang ◽  
Daeui Park
Keyword(s):  
Nitric Oxide ◽  
Polymerase Chain Reaction Analysis ◽  
Inos Mrna ◽  
Raw 264.7 ◽  
Raw 264.7 Macrophages ◽  
Docking Simulations ◽  
Tnf Α ◽  
Cox 2 ◽  
Anti Inflammatory ◽  
Oxide Synthase

We evaluated the anti-inflammatory effects of SNAH in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages by performing nitric oxide (NO) assays, cytokine enzyme-linked immunosorbent assays, Western blotting, and real-time reverse transcription-polymerase chain reaction analysis. SNAH inhibited the production of NO (nitric oxide), reactive oxygen species (ROS), tumor necrosis factor (TNF)-α, and interleukin (IL)-6. Additionally, 100 μM SNAH significantly inhibited total NO and ROS inhibitory activity by 93% (p < 0.001) and 34% (p < 0.05), respectively. Protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) stimulated by LPS were also decreased by SNAH. Moreover, SNAH significantly (p < 0.001) downregulated the TNF-α, IL-6, and iNOS mRNA expression upon LPS stimulation. In addition, 3–100 µM SNAH was not cytotoxic. Docking simulations and enzyme inhibitory assays with COX-2 revealed binding scores of −6.4 kcal/mol (IC50 = 47.8 μM) with SNAH compared to −11.1 kcal/mol (IC50 = 0.45 μM) with celecoxib, a known selective COX-2 inhibitor. Our results demonstrate that SNAH exerts anti-inflammatory effects via suppression of ROS and NO by COX-2 inhibition. Thus, SNAH may be useful as a pharmacological agent for treating inflammation-related diseases.

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