• Molecules and Cells
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• 제24권3호
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• pp.351-357
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• 2007

Regions (about 3.7-3.8 kb) of the mitochondrial genomes (rrnL-cox1) of two tardigrades, a heterotardigrade, Batillipes pennaki, and a eutardigrade, Pseudobiotus spinifer, were sequenced and characterized. The gene order in Batillipes was $\underline{rrnL}-\underline{V}-\underline{rrnS}-\underline{Q}-\underline{I}$-M-nad2-W-$\underline{C}-\underline{Y}$-cox1, and in Pseudobiotus it was $\underline{rrnL}-\underline{V}-\underline{rrnS}-\underline{Q}$-M-nad2-W-$\underline{C}-\underline{Y}$-cox1. With the exception of the trnI gene, the two tardigrade regions have the same gene content and order. Their gene orders are strikingly similar to that of the chelicerate Limulus polyphemus (rrnL-V-rrnS-CR-I-Q-M-nad2-W-C-Y-cox1), which is considered to be ancestral for arthropods. Although the tardigrades do not have a distinct control region (CR) within this segment, the trnI gene in Pseudobiotus is located between rrnL-trnL1 and trnL2-nad1, and the trnI gene in Batillipes is located between trnQ and trnM. In addition, the 106-bp region between trnQ and trnM in Batillipes not only contains two plausible trnI genes with opposite orientations, but also exhibits some CR-like characteristics. The mitochondrial gene arrangements of 183 other protostomes were compared. 60 (52.2%) of the 115 arthropods examined have the M-nad2-W-C-Y-cox1 arrangement, and 88 (76.5%) the M-nad2-W arrangement, as found in the tardigrades. In contrast, no such arrangement was seen in the 70 non-arthropod protostomes studied. These are the first non-sequence molecular data that support the close relationship of tardigrades and arthropods.

• 동의생리병리학회지
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• 제21권2호
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• pp.491-497
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• 2007

Hyeonggaeyeongyo-tang (HYT; Jingjielianqiao-tang), is known to be effective in lowering wind-heat blended as a pathogen of kidney. HYT has been traditionally used for the treatment of a syndrome in kidney meridian, due to invasion of pathogenic wind and heat. Nowadays, this prescription is used to treat diseases marked by excessive wind and heat in the kidney meridian, such as acute otitis media, empyema, hypertrophic rhinitis, nasal bleeding, nasal obstruction, acne and tonsillitis. The present study was conducted to evaluate the effect of HYT on the regulatory mechanism of cytokines and nitric oxide (NO) for the immunological activities in Raw 264.7 cells. After the treatment of HYT water extract, cell viability was measured by MTT assay, NO production was monitored by measuring the nitrite content in culture medium. Cyclooxygenase-2 (COX-2_ and inducible nitric oxide synthase (iNOS) were determined by immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. The production of No was significantly inhibited by pre-treatment (1h) with HYT(0.1-0.3 mg/ml) on LPS-activated Raw264.7 cells. The expression of iNOS and COX-2 protein were up-regulated by LPS, but the increased levels of iNOS and COX-2 were inhibited by pre-treatment of HYT (0.3-1.0 mg/ml), respectively. And the level of interleukin-6 (IL-6), cytokine released from macrophage, was reduced by HYT pre-treatment (0.3-1.0 mg/ml). Thus, the present data suggest that HYT may play an important role in adjunctive therapy in Gram-negative bacterial infections.

• The Korean Journal of Physiology and Pharmacology
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• 제7권4호
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• pp.239-246
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• 2003

The expression of cyclooxygenase-2 (COX-2) is a characteristic response to inflammation and can be inhibited with sodium salicylate. $TNF-{\alpha}$ plus $IFN-{\gamma}$ can induce extracellular signal-regulated kinase (ERK), IKK, $I{\kappa}B$ degradation and NF-${\kappa}B$ activation. The inhibition of the ERK pathway with selective inhibitor, PD098059, blocked cytokine-induced COX-2 expression and $PGE_2$ release. Salicylate treatment inhibited COX-2 expression induced by $TNF-{\alpha}$/$IFN-{\gamma}$ and regulated the activation of ERK, IKK and $I{\kappa}B$ degradation and subsequent NF-${\kappa}B$ activation in MC3T3E1 osteoblasts. Furthermore, antioxidants such as catalase, N-acetyl-cysteine or reduced glutathione attenuated COX-2 expression in combined cytokines-treated cells, and also inhibited the activation of ERK, IKK and NF-${\kappa}B$ in MC3T3E1 osteoblasts. In addition, $TNF-{\alpha}$/$IFN-{\gamma}$ stimulated ROS release in the osteoblasts. However, salicylate had no obvious effect on ROS release in DCFDA assay. The results showed that salicylate inhibited the activation of ERK and IKK, $I{\kappa}B$ degradation and NF-${\kappa}B$ activation independent of ROS release and suggested that salicylate exerts its anti-inflammatory action in part through inhibition of ERK, IKK, $I{\kappa}B$, $NF-{\kappa}B$ and resultant COX-2 expression pathway.

• Archives of Pharmacal Research
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• 제26권10호
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• pp.832-839
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• 2003

Activated macrophages express inducible isoforms of nitric oxide synthase (iNOS) and cyclooxygenase (COX-2), and produce excessive amounts of nitric oxide (NO) and prostaglandin E$_2$ (PGE$_2$), which play key roles in the processes of inflammation and carcinogenesis. The root of Paeonia lactiflora Pall., and the root cortex of Paeonia suffruticosa Andr., are important Chinese crude drugs used in many traditional prescriptions. 1,2,3,4,6-penta-O-galloyl-$\beta$-D-glucose (PGG) is a major bioactive constituent of both crude drugs. PGG has been shown to possess potent anti-oxidant, anti-mutagenic, anti-proliferative and anti-invasive effects. In this study, we examined the inhibitory effects of 1,2,3,4,6-penta-O-galloyl-$\beta$-D-glucose (PGG) isolated from the root of Paeonia lactiflora Pall. on the COX-2 and iNOS activity in LPS-activated Raw 264.7 cells, COX-1 in HEL cells. To investigate the structure-activity relationships of gallate and gallic acid for the inhibition of iNOS and COX-2 activity, we also examined (-)-epigallocatechin gallate (EGCG), gallic acid, and gallacetophenone. The results of the present study indicated that PGG, EGCG, and gallacetophenone treatment except gallic acid significantly inhibited LPS-induced NO production in LPS-activated macrophages. All of the four compounds significantly inhibited COX-2 activity in LPS-activated macrophages. Among the four compounds examined, PGG revealed the most potent in both iNOS ($IC_{50}$ = 18 $\mu\textrm{g}/mL$) and COX-2 inhibitory activity (PGE$_2$: $IC_{50}$ = 8 $\mu\textrm{g}/mL$ and PGD$_2$: $IC_{50}$ = 12 $\mu\textrm{g}/mL$), respectively. Although further studies are needed to elucidate the molecular mechanisms and structure-activity relationship by which PGG exerts its inhibitory actions, our results suggest that PGG might be a candidate for developing anti-inflammatory and cancer chemopreventive agents.

• 한국식품과학회지
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• 제44권6호
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• pp.759-762
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• 2012

이번 실험을 통하여 PEIC가 OVA에 의해 유도된 NF-${\kappa}B$ 활성과 iNOS, COX-2 발현에 어떠한 영향을 미치는지 알아 보았다. PEIC는 OVA에 의해 유도된 NF-${\kappa}B$ 활성을 억제시켰다. 또한 PEIC는 OVA에 의해 유도된 iNOS의 발현도 억제시켰다. 하지만 PEIC는 OVA에 의해 유도된 COX-2 발현은 억제시키지 못하였다. 이러한 결과는 iNOS와 COX-2가 서로 다른 메커니즘에 의해 조절된다는 것을 암시한다. 또한 PEIC는 알러지와 같은 만성적인 질병들을 조절할 수 있는 치료제 개발 및 백신 제조에 중요한 역할을 할 것으로 기대한다.

• 대한한방내과학회지
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• 제29권3호
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• pp.752-764
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• 2008

Objectives : This study was carried out to investigate the effects of Doche-tang on colonic mucosal ulcer induced by dextran sulfate sodium(DSS). Method : The group was divided into three. The normal group consisted of mice that were not inflammation-induced. The control group was composed of untreated colitis elicited mice. The sample group was administered Doche-tang after colitis elicitation. The effects on colonic mucosal ulcers were evaluated by the morphological change of colonic mucosa, the anti-oxidant effect, HSP 70, $NF-{\kappa}B$, COX-1, COX-2 and iNOS. Results : In terms of immunohistochemical findings, the distribution of COX-1 in mice treated with Doche-tang noticeably increased more than that in the control group. The distribution of HSP70, $NF-{\kappa}B$, COX-2, iNOS in mioe treated with Doche-tang decreased more than that in the control group. Regeneration of surface epithelial cell and goblet cell in mucosa was observed by transmission electron microscope. Conclusion : According to the results, Doche-tang is practicable treatment for colonic mucosal ulcer.

• Preventive Nutrition and Food Science
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• 제14권4호
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• pp.269-276
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• 2009

Overproduction of reactive oxygen species (ROS), including nitric oxide (NO), could be associated with the pathogenesis of various diseases such as cancer and chronic inflammation. Inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) are known to play key roles in the development of these diseases. Cedrela sinensis leaves have been used in Asian countries as a traditional remedy for enteritis, dysentery and itching. In the present study, we investigated the anti-inflammatory effects of Cedrela sinensis leaves in lipopolysaccharide (LPS)- stimulated RAW 264.7 macrophages. Powder of C. sinensis leaves was extracted with 95% ethanol and fractionated with a series of organic solvents including n-hexane, dichloromethane, ethyl acetate, n-butanol, and water. The dichloromethane (DCM) fraction strongly inhibited NO production possibly by down-regulating iNOS and COX-2 expression, as determined by Western blotting. Hydrogen peroxide-induced generation of reactive oxygen species (ROS) was also effectively inhibited by the DCM fraction from C. sinensis leaves. In addition, C. sinensis inhibited LPS-mediated p65 activation via the prevention of IκB-$\alpha$ phosphorylation. Furthermore, mitogen-activated protein kinases (MAPKs) such as ERK 1/2 and p38 were found to affect the expression of iNOS and COX-2 in the cells. Taken together, our data suggest that leaves of C. sinensis could be used as a potential source for anti-inflammatory agents.

• 동의생리병리학회지
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• 제30권4호
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• pp.266-273
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• 2016

Ohmae-hwan has been used to treat the symptoms of loose stools mixed with blood accompanied by abdominal pain due to heat dysentery in Korean medicine. However, its mechanism of action on inflammatory disease using macrophages was not known. The author was to find out about the influence of Ohmae-hwan water extract on expressions of NO, cytokine IL-6 and TNF-α in LPS-stimulated RAW 264.7 cell, on expressions of mRNA of iNOS, IL-6 and COX-2 and on suppression of expressions of protein of iNOS and COX-2, and was to find out about the mechanism. As a result of the study, it was found that Ohmae-hwan suppressed generation of NO activated by LPS depending on its concentrations, suppressed expressions of cytokine IL-6, and TNF-α and suppressed expressions of mRNA of iNOS, IL-6, and COX-2 and suppressed expressions of protein of iNOS, COX-2. Considering the above results, it can be known that Ohmae-hwan water extract has anti-inflammatory effect against inflammatory response mediated by macrophages by suppressing generation of NO, TNF-α, and IL-6 and expressions of iNOS, COX-2 as it acts on macrophages.

• Fisheries and Aquatic Sciences
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• 제14권4호
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• pp.275-282
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• 2011

Inflammatory mediators such as inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) have been implicated in various inflammatory diseases. In this study, we investigated the anti-inflammatory activities of Chondria crassicaulis ethanolic extract (CCE) by measuring its effects on the expression of iNOS and COX-2 proteins in lipopolysaccharide (LPS)-treated RAW 264.7 murine macrophages. CCE significantly and dose-dependently inhibited the LPS-induced release of nitric oxide and prostaglandin $E_2$, and suppressed the expression of iNOS and COX-2 proteins in LPS-stimulated RAW 264.7 cells, without causing any cytotoxicity. It also inhibited the production of the pro-inflammatory cytokines such as interleukin (IL)-$1{\beta}$, IL-6, and tumor necrosis factor (TNF)-${\alpha}$ in LPS-stimulated RAW 264.7 cells. Moreover, treatment with CCE strongly suppressed nuclear factor-${\kappa}B$ (NF-${\kappa}B$) promoter-driven expression in LPS-treated RAW 264.7 cells. CCE treatment blocked nuclear translocation of the p65 subunit of NF-${\kappa}B$ by preventing proteolytic degradation of inhibitor of ${\kappa}B-{\alpha}$. These results indicate that CCE regulates iNOS and COX-2 expression through NF-${\kappa}B$-dependent transcriptional control, and identifies potential candidates for the treatment or prevention of inflammatory diseases.

• 생약학회지
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• 제39권2호
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• pp.95-103
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• 2008

In the present study, we investigated the anti-inflammatory effects by kaempferol-3-O-${\beta}$-D-sophoroside (KS) isolated from Sophora japonica (Leguminosae) on the lipopolysaccharide (LPS)-induced nitric oxide (NO) and prostaglandin ($PGE_2$) production by RAW 264.7 cell line compared with kaempferol. KS significantly inhibited the LPS-induced NO and $PGE_2$ production. Consistent with these observations, KS reduced the LPS-induced expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) at the protein and mRNA levels in a concentration-dependent manner. In addition, the release and the mRNA expression levels of tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) and interleukin-6 (IL-6) were also reduced by KS. Moreover, KS attenuated the LPS-induced activation of nuclear factor-kappa B ($NF{-\kappa}B$), a transcription factor necessary for pro-inflammatory mediators, iNOS, COX-2, $TNF-{\alpha}$ and IL-6 expression. These results suggest that the down regulation of iNOS, COX-2, $TNF-{\alpha}$, and IL-6 expression by KS are achieved by the downregulation of $NF{-\kappa}B$ activity, and that is also responsible for its anti-inflammatory effects.