scholarly journals Anti-Inflammatory Activity of 4-((1R,2R)-3-Hydroxy-1-(4-hydroxyphenyl)-1-methoxypropan-2-yl)-2-methoxyphenol Isolated from Juglans mandshurica Maxim. in LPS-Stimulated RAW 264.7 Macrophages and Zebrafish Larvae Model

2021 ◽  
Vol 14 (8) ◽  
pp. 771
Author(s):  
Su-Hyeon Cho ◽  
SeonJu Park ◽  
Hoibin Jeong ◽  
Song-Rae Kim ◽  
Myeong Seon Jeong ◽  
...  

Juglans mandshurica Maxim., a traditional folk medicinal plant, is widely distributed in Korea and China. In our previous study, we isolated a new phenylpropanoid compound, 4-((1R,2R)-3-hydroxy-1-(4-hydroxyphenyl)-1-methoxypropan-2-yl)-2-methoxyphenol (HHMP), from J. mandshurica. In the present study, we evaluated the anti-inflammatory activity of HHMP on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells and zebrafish larvae. HHMP significantly inhibited LPS-induced nitric oxide (NO) and prostaglandin E2 production in a dose-dependent manner. Moreover, HHMP treatment considerably suppressed LPS-induced expression of inducible nitric oxide synthase and cyclooxygenase-2. We also demonstrated the mechanisms of HHMP inhibition of inflammatory responses in LPS-stimulated RAW 264.7 cells via Western blot analysis and immunofluorescence staining. Furthermore, HHMP significantly inhibited NO production in LPS-stimulated zebrafish larvae. Consequently, we established that HHMP significantly inhibited the LPS-induced activation of NF-κB and MAPK and the nuclear translocation of p65 in RAW 264.7 cells. Taken together, our findings demonstrate the effect of HHMP on LPS-induced inflammatory responses in vitro and in vivo, suggesting its potential to be used as a natural anti-inflammatory agent.

Author(s):  
Adek Zamrud Adnan ◽  
Muhammad Taher ◽  
Tika Afriani ◽  
Annisa Fauzana ◽  
Dewi Imelda Roesma ◽  
...  

 Objective: The aim of this study was to investigate in vitro anti-inflammatory activity of tinocrisposide using lipopolysaccharides (LPS)-stimulated RAW 264.7 macrophage cells. Tinocrisposide is a furano diterpene glycoside that was isolated in our previous study from Tinospora crispa.Methods: Anti-inflammatory effect was quantified spectrometrically using Griess method by measuring nitric oxide (NO) production after the addition of Griess reagent.Results: The sample concentrations of 1, 5, 25, 50, and 100 μM and 100 μM of dexamethasone (positive control) have been tested against the LPS-stimulated RAW 264.7 cells, and the results showed NO level production of 39.23, 34.00, 28.9, 20.25, 16.3, and 13.68 μM, respectively, and the inhibition level of 22.67, 33.00, 43.03, 60.10, 68.00, and 73%, respectively.Conclusions: From the study, it could be concluded that tinocrisposide was able to inhibit the formation of NO in the LPS-stimulated RAW 264.7 cells in concentration activity-dependent manner, with half-maximal inhibition concentration 46.92 μM. It can be developed as anti-inflammatory candidate drug because NO is a reactive nitrogen species which is produced by NO synthase. The production of NO has been established as a mediator in inflammatory diseases.


2018 ◽  
Vol 46 (06) ◽  
pp. 1281-1296 ◽  
Author(s):  
Sang Yun Han ◽  
Young-Su Yi ◽  
Seong-Gu Jeong ◽  
Yo Han Hong ◽  
Kang Jun Choi ◽  
...  

Lilium bulbs have long been used as Chinese traditional medicines to alleviate the symptoms of various human inflammatory diseases. However, mechanisms of Lilium bulb-mediated anti-inflammatory activity and the bioactive components in Lilium bulbs remain unknown. In the present study, the anti-inflammatory activity of Lilium bulbs and the underlying mechanism of action were investigated in macrophages using Lilium bulb ethanol extracts (Lb-EE). In a dose-dependent manner, Lb-EE inhibited nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7 cells and bone marrow-derived macrophages (BMDMs) without causing significant cytotoxicity. Lb-EE also down-regulated mRNA expression of inflammatory genes in LPS-stimulated RAW264.7 cells, which included inducuble nitric oxide synthase (iNOS), cyclooxygenase-2 (COX2), and tumor necrosis factor-[Formula: see text] (TNF-[Formula: see text]). Furthermore, Lb-EE markedly restored LPS-induced morphological changes in RAW264.7 cells to a normal morphology. HPLC analysis identified quercetin, luteolin, and kaempferol as bioactive components contained in Lb-EE. Mechanistic studies in LPS-stimulated RAW264.7 cells revealed that Lb-EE suppressed MyD88- and TRIF-induced NF-[Formula: see text]B transcriptional activation and the nuclear translocation of NF-[Formula: see text]B transcription factors. Moreover, Lb-EE inhibited IKK[Formula: see text]/[Formula: see text]-induced activation of the NF-[Formula: see text]B signaling pathway and IKK inhibition significantly reduced NO production in LPS-stimulated RAW264.7 cells. Taken together, these results suggest that Lb-EE plays an anti-inflammatory role by targeting IKK[Formula: see text]/[Formula: see text]-mediated activation of the NF-[Formula: see text]B signaling pathway during macrophage-mediated inflammatory responses.


Author(s):  
Chun Whan Choi ◽  
Ju Young Shin ◽  
Changon Seo ◽  
Seong Su Hong ◽  
Eun-Kyung Ahn ◽  
...  

Background: Plants still remain the prime source of drugs for the treatment of inflammation and can provide leads for the development of novel anti-inflammatory agents. Material and methods: An in vitro bioassay guide revealed that the 80% ethanol (EtOH) extract of the whole plant, Amomum tsao-ko (Zingiberaceae), displayed anti-inflammatory activity after assessing its effects on murine macrophage RAW 264.7 cells. Result: Phytochemical study of the 80% EtOH extract of Amomum tsao-ko led to the isolation of eight compounds: 4-hydroxy-3-methoxy-benzoic acid (1), meso-hannokinol (2), (+)-hannokinol (3), coumaric acid (4), 4-hydroxy-benzoic acid (5), (+)-epicatechin (6), (-)-catechin (7), and myrciaphenone A (8). The results indicated that two of the isolated components, (+)-epicatechin (6) and (-)-catechin (7), inhibited the production of nitric oxide (NO) significantly in lipopolysaccharide treated RAW 264.7 cells. Conclusion: LPS-induced interleukin tumor necrosis factor-alpha (TNF-), IL-1β and IL-10 production was also decreased in a dose-dependent manner. In addition, western blot analysis revealed that (+)-epicatechin (6) and (-)-catechin (7) reduced the expression of inducible nitric oxide synthase and inhibited nuclear localization of nuclear factor kappa-B (NF-κB).


2016 ◽  
Vol 11 (10) ◽  
pp. 1934578X1601101
Author(s):  
Alev Tosun ◽  
Jaemoo Chun ◽  
Igor Jerković ◽  
Zvonimir Marijanović ◽  
Maurizio A. Fenu ◽  
...  

The anti-inflammatory activity of the essential oils from Seseli corymbosum subsp. corymbosum Pall. ex Sm. (SC) and Seseli gummiferum Boiss. & Heldr. subsp. corymbosum (SG) was investigated for the first time on lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. The main constituents (determined by GC-FID and GC-MS analyses) were germacrene D (54.1%) and sabinene (22.4%) in SG oil and β-phellandrene (29.2%), α-phellandrene (8.2%) and germacrene D (2.5%) in SC oil. SC and SG oils inhibited nitric oxide (NO) production with IC50 values of 56.1 and 108.2 μg/mL, respectively. The oils also inhibited prostaglandin E2 (PGE2) with IC50 values of 49.4 μg/mL (SC oil) and 95.5 μg/mL (SG oil). The inhibitory effect of SC and SG oils was accompanied by dose-dependent decreases of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein expressions in LPS-induced RAW 264.7 cells. The research of the reporter gene assay on nuclear factor κB (NF-κB) showed that SC and SG oils inhibited NF-κB transcriptional activity. The obtained results suggest that SC and SG oils exert the anti-inflammatory effects in LPS-stimulated RAW 264.7 cells by suppressing NF-κB activation.


2018 ◽  
Vol 43 (4) ◽  
pp. 375-384 ◽  
Author(s):  
Chang-Gu Hyun ◽  
Min-Jin Kim ◽  
Sang Suk Kim ◽  
Ji Hye Ko ◽  
Young Il Moon ◽  
...  

Abstract Objective In this study, we evaluated the anti-inflammatory effect of Shiranuhi flower in RAW 264.7 cells. Methods The effects of the extracts and solvent fractions on cell viability and LPS-induced inflammatory responses were investigated in RAW 264.7 cells. Results The results showed that the ethyl acetate fraction (HEF) significantly decreased NO production in RAW 264.7 cells; however, cell viability was not affected. In addition, ELISA assay revealed that HEF significantly inhibited the productions of PGE2, TNF-α, and IL-6. As well, using Western blot analysis, it was observed that HEF significantly reduced the expression levels of iNOS and COX-2 in a dose dependent manner. Furthermore, we detected a reduced phosphorylation of mitogen-activated protein kinases such as p38, JNK, and ERK1/2. This indicates that HEF regulates LPS-induced inflammatory responses, at least in part, via suppressing the MAPK signaling pathway. Correlation analysis also showed that anti-inflammatory activities were highly correlated to antioxidant activities in this study. Characterization of the Shiranuhi flowers for flavonoid contents using HPLC showed varied quantity of narirutin and hesperidin. Conclusion Overall, the results demonstrate that HEF may be a potential anti-inflammatory agent. In addition, our findings contribute to understanding the molecular mechanism underlying the anti-inflammatory effect of Shiranuhi flower.


Molecules ◽  
2020 ◽  
Vol 25 (17) ◽  
pp. 3784
Author(s):  
Jingya Ruan ◽  
Ying Zhang ◽  
Wei Zhao ◽  
Fan Sun ◽  
Lifeng Han ◽  
...  

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.


Molecules ◽  
2019 ◽  
Vol 24 (2) ◽  
pp. 275 ◽  
Author(s):  
Fanhui Kong ◽  
Bae Lee ◽  
Kun Wei

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.


2015 ◽  
Vol 77 (25) ◽  
Author(s):  
Siti Aisha Mohd Radzi ◽  
Yosie Andriani ◽  
Habsah M. ◽  
Tengku Sifzizul Tengku Mohamad ◽  
Jasnizat Saidin

In recent years, high numbers of research focusing on secondary metabolites produced by terrestrial bacteria towards anti-inflammatory properties but, minority of them used bacteria associated to marine organism. In this study, 25 bacteria colonies were isolated from sponge Theonella sp. which was collected from Bidong Island, Terengganu. Cytotoxicity and anti-inflammatory activity of secondary metabolites produced by these bacteria were studied using macrophage cell lines: RAW 264.7 .Macrophage plays a role in mammalian immune system by providing defense against a foreign harmful substance for example Lipopolysaccharides (LPS) which induces production of pro-inflammatory mediators such as nitric oxide (NO). Inhibition of NO production in LPS stimulated RAW 264.7 cells is highly established assay to screen for anti-inflammatory activity. All isolates were cultured and supernatants were collected for the extraction of secondary metabolites using diaion HP-20 to obtain crude extracts. Evaluations of the cytotoxicity effect of the extracts on RAW 264.7 were done using MTT assay. The highest concentration sample producing lowest toxicity effect was used for anti-inflammatory assay. Macrophages were induced with LPS before treated with different concentrations of crude metabolites in anti-inflammatory assay. The effects of crude secondary metabolites extracted to the induced macrophage were monitored by observing the inhibition level of Nitric Oxide (NO) released by RAW 264.7 into supernatant and estimated from nitrate standard curve. A preliminary assay revealed that isolate label with TM1.8 and TM 1.9 possesses anti-inflammatory activities of 112.06% and 109.7% respectively for every 1ug/L sample. The result suggested that these two crude extracts were more effective in inhibiting NO production compared to positive control N-Monomethyl-L-arginine Monoacetate (L-NMMA) which showed an inhibitory activity at 87.41% for every 1ug/L sample.


Author(s):  
Krishna Chaithanya K ◽  
Gopalakrishnan V K ◽  
ZenebeHagos . ◽  
Nagaraju B ◽  
Kamalakararao K ◽  
...  

Objective: Mesuaferrea L is a medicinal plant belongs to the family Clusiace, it is extensively used in folk medicine for treatment of chronic inflammatory diseases.The present study was aimed to evaluate in vitro and in vivo anti-inflammatory activity of M. ferrea L. Methods: The in vitro anti-inflammatory activities such as nitric oxide, PGE2, pro-inflammatory cytokines (TNF-α and IL-1β) were studied in RAW 264.7 cells and in vivo studies were carried out on carrageenan -induced inflammation in Wistar rats. The sequentially extracted M. ferreaL bark extracts (MFBHE, MFBEE, and MFBME) exhibited inhibitory effects on pro-inflammatory mediators such as nitric oxide, prostaglandin E2, tumour necrosis factorαandinterleukin-1βproduction in concentration dependent manner in LPS induced RAW 264.7 cells andCarrageenan induced paw oedema in Wistar rats. Conclusion: The result of the present study indicated that M. ferrea L ethyl acetate bark extract exhibited significant in vitroand in vivoanti-inflammatory activity.


Molecules ◽  
2019 ◽  
Vol 24 (21) ◽  
pp. 3910 ◽  
Author(s):  
Min-Seon Kim ◽  
Jin-Soo Park ◽  
You Chul Chung ◽  
Sungchan Jang ◽  
Chang-Gu Hyun ◽  
...  

Biorenovation is a microbial enzyme-catalyzed structural modification of organic compounds with the potential benefits of reduced toxicity and improved biological properties relative to their precursor compounds. In this study, we synthesized a novel compound verified as formononetin 7-O-phosphate (FMP) from formononetin (FM) using microbial biotransformation. We further compared the anti-inflammatory properties of FMP to FM in lipopolysaccharide (LPS)-treated RAW264.7 macrophage cells. We observed that cell viabilities and inhibitory effects on LPS-induced nitric oxide (NO) production were greater in FMP-treated RAW 264.7 cells than in their FM-treated counterparts. In addition, FMP treatment suppressed the production of proinflammatory cytokines such as prostaglandin-E2 (PGE2), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in a dose-dependent manner and concomitantly decreased the mRNA expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2). We also found that FMP exerted its anti-inflammatory effects through the downregulation of the extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and nuclear factor kappa B (NF-κB) signaling pathways. In conclusion, we generated a novel anti-inflammatory compound using biorenovation and demonstrated its efficacy in cell-based in vitro assays.


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