• Biomolecules & Therapeutics
• /
• v.21 no.2
• /
• pp.146-152
• /
• 2013

This study examined the total polyphenol content of eight wild edible plants from Ethiopia and their effect on NO production in Raw264.7 cells. Owing to its relatively high polyphenol concentration and inhibition of NO production, the methanol extract of Adansonia digitata L. leaf (MEAD) was subjected to detailed evaluation of its antioxidant and anti-inflammatory effects. Antioxidant effects were assessed by measuring free-radical-scavenging activity using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and oxygen-radical-absorbance capacity (ORAC) assays, while anti-inflammatory effects were assessed by measuring inducible nitric oxide synthase (iNOS) expression in lipopolysaccharide (LPS)-stimulated RAW264.7 cells. In the ORAC assay, MEAD was 10.2 times more potent than vitamin C at eliminating peroxyl radicals. In DPPH assay, MEAD also showed a strong ROS scavenging effect. MEAD significantly inhibited iNOS activity ($IC_{50}=28.6{\mu}g/ml$) of LPS-stimulated Raw264.7 cells. We also investigated the relationship between iNOS expression and nuclear factor kappa B (NF-${\kappa}B$) activation. MEAD inhibited $I{\kappa}B{\alpha}$ degradation and NF-${\kappa}B$ translocation from the cytosol to the nucleus in LPS-induced RAW264.7 cells without significant cytotoxic effects, as confirmed by MTT assay. These results suggest that MEAD inhibits anti-inflammatory iNOS expression, which might be related to the elimination of peroxyl radicals and thus the inhibition of $I{\kappa}B{\alpha}$-mediated NF-${\kappa}B$ signal transduction.

• Journal of the Korea Academia-Industrial cooperation Society
• /
• v.15 no.8
• /
• pp.5088-5094
• /
• 2014

Hovenia dulcis extract (HDE) has positive effects on alcohol degradation, recovery of liver damage and antioxidant activities. This study examined whether HDE exerts an ameliorative effect on inflammatory orifacial pain in an animal algesic model with formalin. The animals (rats) were divided into four groups: group I (control), group II (right facial subcutaneous injection of 5% formalin, inflammatory orifacial pain group), group III (5% formalin + distilled water administration), and group IV injection (5% formalin + 4.5 ml/kg of HDE), respectively. The scores from the scratch and effleurage tests were applied to evaluate the differences between three groups. The expression of p38 MAPK, iNOS and Nrf2 in the brain and medulla oblongata, which are involved in pain regulation, inflammation, antioxidation and nitric oxide production, were analyzed by western blot. The degree of orifacial pain was significantly lower in group IV than in groups I, II and, III. The expression of p38MAPK, iNOS and Nrf2 in the brain and medulla were also lower in group IV than in the other groups. These findings suggested that a Hovenia dulcis extract can attenuate inflammatory orifacial pain by suppressing the expression of p38 MAPK, iNOS and Nrf2.

• Korean Journal of Poultry Science
• /
• v.40 no.2
• /
• pp.83-89
• /
• 2013

iNOS (Inducible nitric oxide synthase) and TLR-4 (Toll-like Receptor-4) play crucial roles in innate immunity of poultry. iNOS has been mapped to chicken chromosome 14 and implicated in a variety of chicken diseases. iNOS possesses potent antimicrobial activity, including the inhibition of microbes replication in vitro. TLR-4 is a pathogen associated molecular-pattern receptor for bacterial product, such as LPS (lipopolysaccharides) found in Gram negative bacteria, that triggers pro-inflammatory cytokine expression after engagement with ligands. In the previous studies, genetic analysis of iNOS and TLR-4 revealed the possible association of mutation in these genes with the intestinal microflora of cecum when infected with Salmonella spp. This study was aimed to augment previous findings, which show the association of iNOS (C14513T) and TLR-4 (G4409T) polymorphisms with economic traits in Korean Native Black (KNB), Rhode Island Red (RIR) and Cornish chickens. Investigation in the effect of SNPs on economic traits (layday, layw, layno, bw150, bw270, layw270) was conducted. iNOS (C14513T) had a significant effect on the average body weight at 270 days of age (p<0.05) in both KNB and RIR, whereas TLR-4 (G4409T) showed no significant correlation with all traits (p>0.05). The results obtained from using the candidate genes can be useful for the genetic improvement of body weight in both KNB and RIR breeds.

• Journal of Acupuncture Research
• /
• v.29 no.1
• /
• pp.15-24
• /
• 2012

Objectives : In recent years, many studies have been widely researching anti-inflammation effect of various medicinal plants. $Cinnamomi$ $Cortex$ was not enough in researching of the anti-inflammation. Moreover, there is no comparative study about extraction methods. Therefore, we investigated the inhibitory effects of $Cinnamomi$ $Cortex$ pharmacopuncture by EtOH and Hot water extraction on Nitric oxide(NO), Prostaglandin E2(PGE2) production, Cyclooxygenase(COX)-2, inducible NOS(iNOS) expression and extracellular signal regulate kinase(ERK)1/2 phosphorylation in lipopolysaccharide(LPS) induced RAW 264.7 macrophage cell. Methods : $Cinnamomi$ $Cortex$ was extracted by EtOH and Hot water. RAW 264.7 macrophage cell viability was measured by MTT assay. Effect of $Cinnamomi$ $Cortex$ pharmacopuncture on NO and PGE2 production in LPS induced macrophages was accessed by Griess assay and enzyme-linked immunospecific assay(ELISA), respectively. Inhibition effect on COX-2, iNOS expression and ERK1/2 phosphorylation was examined by Immunoblotting assay. Results : 1. Cytotoxic effect of $Cinnamomi$ $Cortex$ pharmacopuncture by Hot water extraction in RAW 264.7 macrophages was not appeared, except $3125{\mu}g/m{\ell}$. And cytotoxic effect was not appeared in EtOH extraction method. 2. $Cinnamomi$ $Cortex$ pharmacopuncture by EtOH and Hot water extraction inhibited NO production in LPS induced macrophages significantly. 3. $Cinnamomi$ $Cortex$ pharmacopuncture by EtOH and Hot water extraction inhibited PGE2 production in LPS induced macrophages significantly. 4. $Cinnamomi$ $Cortex$ pharmacopuncture by EtOH and Hot water extraction inhibited COX-2, iNOS expression in LPS induced macrophages. Especially, it has been confirmed that COX-2, iNOS expression were effectively inhibited in Hot water extraction. 5. $Cinnamomi$ $Cortex$ pharmacopuncture by EtOH and Hot water extraction inhibited ERK1/2 phosphorylation in LPS induced macrophages. Especially, it has been confirmed that ERK1/2 phosphorylation was effectively inhibited in Hot water extraction. Conclusions : According to the results, $Cinnamomi$ $Cortex$ pharmacopuncture suppresses NO, PGE2 production, COX-2, iNOS expression and ERK1/2 phosphorylation in LPS induced macrophages. It has a potential for treating various inflammatory diseases, and Hot water extraction method could be used more extensively than EtOH extraction method.

• Food Science and Biotechnology
• /
• v.15 no.2
• /
• pp.270-276
• /
• 2006

We evaluated the effects of extracts of Tsunokaori tangor peel on lipopolysaccharide (LPS)-induced nitric oxide (NO) and prostaglandin $E_2\;(PGE_2)$ in RAW 264.7 cells. The ethyl acetate fraction of Tsunokaori tangor peel (EA-TTP) markedly inhibited the production of NO and $PGE_2$ in LPS-stimulated RAW 264.7 cells. Consistent with these findings, the expression levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) proteins were down-regulated in a dose-dependent manner. Additionally, EA-TTP decreased the expression iNOS mRNA but not COX-2 mRNA. To determine the upstream signaling mechanism for the down-regulation of LPS-induced iNOS expression, we investigated the effect of EA-TTP on the degradation and re-synthesis of $I{\kappa}B{\alpha}$. EA-TTP dose-dependently delayed $I{\kappa}B{\alpha}$ degradation and increased $I{\kappa}B{\alpha}$ re-appearance following degradation, suggesting this as the mechanism by which EA-TTP suppressed iNOS gene expression. The EA-TTP also dose-dependently reduced the expression of the cellular stress-response protein heme oxygenase-1, and inhibited the LPS-induced sustained activation of extracellar signal-regulated kinase (ERK).

• Tuberculosis and Respiratory Diseases
• /
• v.60 no.4
• /
• pp.426-436
• /
• 2006

Background : Particulate matters (PM) when inhaled is known to induce pulmonary diseases including asthma and chronic bronchitis when inhaled. Despite the epidemiological proofevidence, the pathogenesis of PM-related pulmonary diseases is unclearremain poorly understood. Methods : Primary alveolar macrophages were harvested from the SPF and inflammatory rats by bronchioalveolar lavage (BAL). The cultured primary alveolar macrophages were treated with the medium only, PM only ($5{\sim}40{\mu}g/cm^2$), LPS (5ng/ml) only, and PM with LPS for 24 and 48 hours. The level of secreted nitric oxide (NO) was assayed from the cultured medium by using the Griess reaction. The cultured cells were utilized for the western blotting against the inducible nitric oxide synthase (iNOS) proteins. Immunocyto- chemical staining against the iNOS and NT-proteins were performed in cells that cultured in the $Lab-Tek^{(R)}$ chamber slide after treatments. Results : The PM that utilizein this experiments induced NO formation with iNOS expression in the cultured SPF and inflammatory rats alveolar macrophages, by itself. When the cells were co-treated with PM and LPS, there was a statistically significant synergistic effect on NO formation and iNOS expression over the LPS effect. The cells from the sham control showed minimal immunoreactivity for the NT-proteins. Significantly higher quantities of NT-proteins were detected in the PM and PM with LPS co-treated cells than from the sham control. Conclusion : Increased iNOS expression and NO formation with increased NT-proteins formation might be involved in the pathogenesis of PM-induced lung injury.

• The Korean Journal of Physiology and Pharmacology
• /
• v.8 no.5
• /
• pp.273-280
• /
• 2004

Nitric oxide (NO) has been suggested to act as a mediator of cytokine-induced effects of turn over of bone. Activation of the inducible nitric oxide synthase (iNOS) by inflammation has been related with apoptotic cell death in osteoblast. YS 49, a synthetic isoquinoline alkaloid, inhibits NO production in macrophages activated with cytokines. In the present study, we investigated the molecular mechanism of YS 49 to inhibit iNOS expression in ROS 17/2.8 cells, which were activated with combined treatment of inflammatory cytokines $(TNF-{\alpha},\;IFN-{\gamma})$ and lipopolysaccharide (LPS). Results indicated that YS 49 concentration-dependently reduced iNOS mRNA and protein expression, as evidenced by Northern and Western blot analysis, respectively. The underlying mechanism by which YS 49 suppressed iNOS expression was not to affect iNOS mRNA stability but to inhibit activation and translocation of $NF-_kB$ by preventing the degradation of its inhibitory protein $I_kB_{\alpha}$. As expected, YS 49 prevented NO-induced apoptotic cell death by sodium nitroprusside. Taken together, it is concluded that YS 49 inhibits iNOS expression by interfering with degradation of phosphorylated inhibitory $_kB_{\alpha}\;(p-I_kB_{\alpha})$. These actions may be beneficial for the treatment of inflammation of the joint, such as rheumatoid arthritis.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.48 no.3
• /
• pp.213-223
• /
• 2022

In this study, the anti-inflammatory and antioxidant effects of aerial parts of Rumex acetosa L. extract were confirmed to prevent various inflammatory diseases and skin aging caused by excessive oxidative stress. As a result of ABTS assay, it was confirmed that the radical scavenging ability increased in a concentration-dependent manner. ROS inhibitory ability was confirmed through DCF-DA assay, and concentration-dependent inhibition of ROS production was confirmed. The effect of inhibiting cell nuclear damage according to ROS was confirmed through DAPI staining. In addition, it was confirmed that the mRNA expression levels of iNOS and COX-2 were inhibited in a concentration-dependent manner through qPCR. As a result of confirming the protein levels of iNOS and COX-2 by western blotting, iNOS was significantly decreased at all concentrations, and COX-2 was significantly decreased at 800 ㎍/mL. The inhibitory effect on the production of NO generated by iNOS was confirmed by NO assay, and NO was decreased in a concentration-dependent manner. In addition, phosphorylation of ERK and JNK in the MAPKs signaling pathway were inhibited. Therefore, Rumex acetosa L. has the potential to be used as an anti-inflammatory and antioxidant cosmetic raw material by showing anti-inflammatory and antioxidant effects through the MAPKs pathway.

• Journal of The Korean Association For Science Education
• /
• v.36 no.1
• /
• pp.147-158
• /
• 2016

This study examined the changes of preservice biology teachers' NOS pedagogical content knowledge through two consecutive science methods courses: NOS understandings; attitudes toward teaching science; difficulties of NOS teaching; NOS teaching strategies; and views of orientation of NOS teaching. During the science methods course I, the preservice teachers engaged in discussions and reflections on what science is and how scientific knowledge has produced, drawing NOS aspects from episodes of history of science, and planning the lessons cooperating NOS instructional objectives. Then the next semester, through the science methods course II, the preservice teachers had a chance of the simulated teaching by adopting NOS teaching and learning activities in the context of the secondary biology context. The preservice teachers, further, reflected on their NOS teaching. The results showed that the preservice teachers constructed the NOS pedagogical content knowledge. They significantly improved their views of NOS and its teaching(p<.05) after the science methods course I, and retained their understanding after the science methods course II(p>.05). The preservice teachers mentioned the difficulties of teaching NOS in the secondary biology context, and further suggested effective NOS teaching methods in their reflective journals.

• Biomedical Science Letters
• /
• v.14 no.2
• /
• pp.127-130
• /
• 2008

To investigate if there are tandem repeats in iNOS gene in Korean genome we applied PCR amplification followed by DNA sequencing. Tandem repeats we were looking at were (AAAT)n in the promoter region. Totally, 65 people were subjected for this experiment. Twenty of them were patients with metabolic disease. Only $(AAAT)_4$ was found in all of these Korean samples. This result was somewhat different trom the data for Caucasians and other Asian people. So, we assume this is specific VNTR (variable number of tandem repeat) in Korean which can be used for the purpose of diagnosis and for the differentiation of ethnic groups.