• Biomolecules & Therapeutics
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• 제24권6호
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• pp.581-588
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• 2016

Lonchocarpine is a phenylpropanoid compound isolated from Abrus precatorius that has anti-bacterial, anti-inflammatory, antiproliferative, and antiepileptic activities. In the present study, we investigated the antioxidant effects of lonchocarpine in brain glial cells and analyzed its molecular mechanisms. We found that lonchocarpine suppressed reactive oxygen species (ROS) production and cell death in hydrogen peroxide-treated primary astrocytes. In addition, lonchocarpine increased the expression of anti-oxidant enzymes, such as heme oxygenase-1 (HO-1), NAD(P)H:quinone oxidoreductase 1 (NQO1), and manganese superoxide dismutase (MnSOD), which are all under the control of Nrf2/antioxidant response element (ARE) signaling. Further, mechanistic studies showed that lonchocarpine increases the nuclear translocation and DNA binding of Nrf2 to ARE as well as ARE-mediated transcriptional activities. Moreover, lonchocarpine increased the phosphorylation of AMP-activated protein kinase (AMPK) and three types of mitogen-activated protein kinases (MAPKs). By treating astrocytes with each signaling pathway-specific inhibitor, AMPK, c-jun N-terminal protein kinase (JNK), and p38 MAPK were identified to be involved in lonchocarpine-induced HO-1 expression and ARE-mediated transcriptional activities. Therefore, lonchocarpine may be a potential therapeutic agent for neurode-generative diseases that are associated with oxidative stress.

• Journal of Ginseng Research
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• 제32권4호
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• pp.315-323
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• 2008

In the previous studies, we isolated the compound K rich fractions (CKRF) and showed that CKRF inhibited Toll-like receptor (TLR) 4- or TLR9-induced inflammatory signaling. To extend our previous studies,1) we investigated the molecular mechanisms of CKRF in the TLR4-associated signaling via nuclear factor (NF)-${\kappa}B$, and in vivo role of CKRF for induction of tolerance in lipopolysaccharide (LPS)-induced septic shock. In murine bone marrow-dervied macrophages, CKRF significantly inhibited the induction of mRNA expression of proinflammatory mediators such as tumor necrosis factor-${\alpha}$, interleukin-6, cyclooxygenase-2, and inducible nitric oxide synthase. In addition, CKRF significantly attenuated the transcriptional activities of TLR4/LPS-induced NF-${\kappa}B$. Nuclear translocation of NF-${\kappa}B$ in response to LPS stimulation was significantly abrogated by pre-treatment with CKRF. Furthermore, CKRF inhibited the recruitment of p65 to the interferon-sensitive response element flanking region in response to LPS. Finally, oral administration of CKRF significantly protected mice from Gram-negative bacterial LPS-induced lethal shock and inhibited systemic inflammatory cytokine levels. Together, these results demonstrate that CKRF modulates the TLR4-dependent NF-${\kappa}B$ activation, and suggest a therapeutic role for Gram-negative septic shock.

• 대한한의학방제학회지
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• 제24권4호
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• pp.259-269
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• 2016

Objectives : Phyllostachys Folium is leaves of Phyllostachys nigra var. henesis $S_{TAPF}$. In the East Asian traditional medicine, the herb has been used to treat nasal bleeding, dysuria, epilepsy and etc. The present study was conducted to evaluate the anti-inflammatory effects of the Phyllostachys Folium water extracts (PFE) in vitro and in vivo model. Methods : Cell viability was measured by MTT assay after the treatment of PFE and NO production was monitored by measuring the nitrite content in culture medium. iNOS, COX-2, $I{\kappa}B$, $p-I{\kappa}B{\alpha}$ amd $NF{\kappa}B$ were detected by immunoblot analysis, and levels of cytokine were analyzed by sandwich ELISA kit. Anti-edema effect of PFE was determined in the carrageenan-induced paw edema model in rats. Results : LPS increased NO and cytokines levels compared with control, these increases were attenuated by PFE. In addition, LPS-induced pro-inflammatory proteins such as iNOS, COX-2 were down regulated by PFE. These anti-inflammatory effect of PFE results from inhibition of phosphorylation of $I{\kappa}B$ and translocation of $NF-{\kappa}B$. Conclusion : These results show that PFE has some anti-inflammatory effects which might play a role in gram-negative bacterial infection inflammation and $NF{\kappa}B$ activated diseases.

• Journal of Microbiology and Biotechnology
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• 제15권3호
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• pp.603-608
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• 2005

We recently isolated a novel probiotic strain, Lactobacillus fermentum PL9005 (KCCM-10250), from infant feces and showed that it had a potential immunoenhancing effect. In the present study, a safety assessment of the bacteria was performed using a BALB/c mouse model. Mice were administered with L. fermentum PL9005 daily for 28 days. There were no detectable changes in body weight, feed intake, or clinical signs, and no significant difference in hematological parameters or blood biochemistry between the L. fermentum PL9005-fed and control groups. Bacterial translocation was detected in the mesenteric lymph nodes, liver, and spleen of some mice with and without L. fermentum PL9005 feeding, however, the organisms were not related to ingestion of L. fermentum PL9005; this was confirmed by PCR using a species-specific primer. No gross lesions were detected in the liver, spleen, or intestine of L. fermentum PL9005-fed or control mice. Mucosal thickness in the ileum, cecum, and colon of L. fermentum PL9005-fed mice was not significantly different from that of corresponding organs in control mice. No inflammation or epithelial cell degeneration in the intestines was observed in any mice. These results indicate that ingestion of L. fermentum PL9005 is safe in mice and can be applied in the functional food market.

• The Korean Journal of Physiology and Pharmacology
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• 제19권5호
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• pp.441-449
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• 2015

Flavonoids are plant pigments that have been demonstrated to exert various pharmacological effects including anti-cancer, anti-diabetic, anti-atherosclerotic, anti-bacterial, and anti-inflammatory activities. However, the molecular mechanisms in terms of exact target proteins of flavonoids are not fully elucidated yet. In this study, we aimed to evaluate the anti-inflammatory mechanism of scutellarein (SCT), a flavonoid isolated from Erigeron breviscapus, Clerodendrum phlomidis and Oroxylum indicum Vent that have been traditionally used to treat various inflammatory diseases in China and Brazil. For this purpose, a nitric oxide (NO) assay, polymerase chain reaction (PCR), nuclear fractionation, immunoblot analysis, a kinase assay, and an overexpression strategy were employed. Scutellarein significantly inhibited NO production in a dose-dependent manner and reduced the mRNA expression levels of inducible NO synthase (iNOS) and tumor necrosis factor (TNF)-${\alpha}$ in lipopolysaccharide (LPS)-activated RAW264.7 cells. In addition, SCT also dampened nuclear factor (NF)-${\kappa}B$-driven expression of a luciferase reporter gene upon transfection of a TIR-domain-containing adapter-inducing interferon-${\beta}$ (TRIF) construct into Human embryonic kidney 293 (HEK 293) cells; similarly, NF-${\kappa}B$ nuclear translocation was inhibited by SCT. Moreover, the phosphorylation levels of various upstream signaling enzymes involved in NF-${\kappa}B$ activation were decreased by SCT treatment in LPS-treated RAW264.7 cells. Finally, SCT strongly inhibited Src kinase activity and also inhibited the autophosphorylation of overexpressed Src. Therefore, our data suggest that SCT can block the inflammatory response by directly inhibiting Src kinase activity linked to NF-${\kappa}B$ activation.

• 문화기술의 융합
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• 제9권1호
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• pp.717-724
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• 2023

사플라워와 마더워트 등 에탄올복합추출물(SEC)을 lipopolysaccharide(LPS)로 유도된 마우스 대식세포주인 RAW264.7 세포와 마우스 폐포 대식세포주인 MH-S 세포에서 사플라워와 마더워트 등 에탄올복합추출물((SEC)의 항균과 항염증 활성 효과를 조사하였다. 그 결과 SEC(500 ㎍/mL)의 전처리 LPS 자극 세포에서 iNOS 단백질 및 염증성 사이토카인 mRNA 발현을 크게 억제하였다. 사플라워와 마더워트 등 에탄올복합추출물(SEC)에 의한 항염증활성 효과는 다음과 같이 관찰되었다. 사플라워와 마더워트 등 에탄올복합추출물(SEC)은 IκB-α 인산화의 억제를 통해 시토졸에서 핵으로의 NF-κB의 전좌를 억제하고 또한 LPS로 자극된 NF-κB 전사 활성을 억제하였다. 이러한 결과는 사플라워와 마더워트 등 에탄올복합추출물(SEC)이 항염증 작용을 발휘 하고 사플라워와 마더워트 등 에탄올복합추출물(SEC)의 잠재적인 치료 가치성 및 기본 메커니즘을 규명 할 수 있었다. 결론적으로 사플라워와 마더워트 등 에탄올복합추출물(GRP)이 다양한 항염증 활성성분 함유 및 천연 항염증제의 잠재적 공급원으로 활용 가능성이 있음을 시사 하였다.

• 대한한의학방제학회지
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• 제17권2호
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• pp.151-162
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• 2009

Herba Artemisiae Capillaris is the dried bud of Artemisia capillaris Thunb, which has been used for expelling heat to loosen the bowels and normalizing gallbladder function to cure jaundice in traditional oriental medicines. In the present study, we evaluated the anti-inflammatory effects of the aqueous extracts of Herba Artemisiae Capillaris (HAC) in LPS-activated Raw 264.7 cells. Cells were treated with $1\;{\mu}g/ml$ of LPS 1 h before adding HAC extract. Cell viability was determined by MTT assay, and the relative level of NO was measured with Griess reagent. TNF-$\alpha$, IL-$1{\beta}$, and IL-6 cytokines were detected by ELISA. During the entire experimental period, all three doses of HAC extract (0.03, 0.10 and 0.30 mg/ml) had no significant cytotoxicity. LPS-activated cells showed increased NO levels and iNOS expressions compared to control. However, these increases were dramatically attenuated by treatment with HAC extract. Moreover, the inhibitory effects of HAC extract occurred in a dose-dependent manner. In addition, HAC extract reduced the translocation of $NF{\kappa}B$ into nuclear. HAC reduced production of IL-$1{\beta}$ and IL-6 by LPS, although it had no effects on TNF-$\alpha$. These results demonstrate that liquiritigenin exerts anti-inflammatory effects, which results from the inhibition of $NF{\kappa}B$ activation in macrophages, thereby decreasing production of iNOS and proinflammatory cytokines. Taken together, these results indicate that the aqueous extracts of Herba Artemisiae Capillaris warrant further development as an anti-inflammatory agent for the treatment of gram-negative bacterial infections.

• Fisheries and Aquatic Sciences
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• 제14권4호
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• pp.267-274
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• 2011

Bacterial lipopolysaccharide (LPS) induces expression of pro-inflammatory cytokines and enzymes producing nitric oxide (NO) and prostaglandins (PGs) in immune cells. This process is mediated by the activation of nuclear factor kappaB (NF-${\kappa}B$). In this study, we investigated the anti-inflammatory characteristics of Codium fragile ethanolic extract (CFE) mediated by the regulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) using LPS-stimulated murine macrophage RAW 264.7 cells. CFE significantly inhibited LPS-induced NO and $PGE_2$ production in a dose-dependent manner and suppressed the expression of iNOS and COX-2 proteins in LPS-stimulated RAW 264.7 cells with no cytotoxicity. Pro-inflammatory cytokines, such as interleukin (IL)-$1{\beta}$, IL-6, and tumor necrosis factor-${\alpha}$, were significantly reduced by treatment of CFE in LPS-stimulated RAW 264.7 cells. CFE inhibited the promoter activity of (NF)-${\kappa}B$ in LPS-stimulated macrophages. Treatment with CFE suppressed translocation of the NF-${\kappa}B$ p65 subunit by preventing proteolytic degradation of inhibitor of ${\kappa}B-{\alpha}$. These results indicate that the CFE-mediated inhibition of NO and $PGE_2$ production in LPS-stimulated RAW 264.7 cells is mediated through the NF-${\kappa}B$-dependent transcriptional downregulation of iNOS and COX-2, suggesting the potential of CFE as a nutraceutical with anti-inflammatory activity.

• Journal of Microbiology and Biotechnology
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• 제25권10호
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• pp.1751-1760
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• 2015

Mucin2 (MUC2), an important regulatory factor in the immune system, plays an important role in the host defense system against bacterial translocation. Probiotics known to regulate MUC2 gene expression have been widely studied, but the interactions among probiotic, pathogens, and mucin gene are still not fully understood. The aim of this study was to investigate the role of MUC2 in blocking effects of probiotics on meningitic E. coli-induced pathogenicities. In this study, live combined probiotic tablets containing living Bifidobacterium, Lactobacillus bulgaricus, and Streptococcus thermophilus were used. MUC2 expression was knocked down in Caco-2 cells by RNA interference. 5-Aza-2'-deoxycytidine (5-Aza-CdR), which enhances mucin-promoted probiotic effects through inducing production of Sadenosyl-L-methionine (SAMe), was used to up-regulate MUC2 expression in Caco-2 cells. The adhesion to and invasion of meningitic E. coli were detected by competition assays. Our studies showed that probiotic agents could block E. coli-caused intestinal colonization, bacteremia, and meningitis in a neonatal sepsis and meningitis rat model. MUC2 gene expression in the neonatal rats given probiotic agents was obviously higher than that of the infected and uninfected control groups without probiotic treatment. The prohibitive effects of probiotic agents on MUC2-knockdown Caco-2 cells infected with E44 were significantly reduced compared with nontransfected Caco-2 cells. Moreover, the results also showed that 5-Aza-CdR, a drug enhancing the production of SAMe that is a protective agent of probiotics, was able to significantly suppress adhesion and invasion of E44 to Caco-2 cells by upregulation of MUC2 expression. Taken together, our data suggest that probiotic agents can efficiently block meningitic E. coli-induced pathogenicities in a manner dependent on MUC2.

• 생명과학회지
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• 제24권6호
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• pp.664-670
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• 2014

비브리오균(Vibrio vulnificus)은 심각하고 치명적인 감염을 일으킬 수 있는 호염성의 식중독균이지만, 숙주세포 내에서 염증반응을 일으키는 분자적 기작은 아직 잘 알려지지 않았다. 본 연구에서는 오염된 식품 섭취를 통해 유입되는 비브리오균의 소장 특이적 염증 반응 위치와 기작을 알아보기 위해, 7주령의 수컷 마우스에 비브리오균($1{\times}10^9CFU$)을 16시간 동안 경구 투여하였다. 그 결과 비브리오균은 주로 회장(ileum) 부위에서 비브리오균(WT) 수가 가장 많이 증가하였고, 공장(Jejunum), 근위부대장(proximal colon), 원위부 대장(distal colon)에도 유의적으로 군집현성이 이루어짐을 알 수 있었지만, 십이지장(duodenum)과 비장(spleen), 그리고 간(liver) 조직들에서는 관찰되지 않았다. 특히 비브리오균의 표적기관인 회장상피조직에서는 비브리오균(WT)이 침입 시 융모 안으로 다량의 염증세포들이 유입되었고, 융포의 폭이 넓어지고 길이가 짧아지는 전형적인 조직학적 염증 반응을 보여주었다. 비브리오균이 유도한 조직 특이적 염증반응기작을 알아보기 위해, 비브리오에 감염된 회장상피조직으로부터 단백질과 mRNA를 분리하였다. 비브리오균은 숙주세포의 중추적 신호전달 단백질인 protein kinase C (PKC)의 인산화 및 $PKC{\alpha}$의 세포막이동을 촉진시켰고, mitogen-activated protein kinase (MAPK) 중 extracellular signal-regulated kinases (ERK)와 c-Jun N-terminal kinases (JNK)의 인산화를 유도하였지만, p38 MAPK 인산화에는 영향을 미치지 않았다. 특히 비브리오균은 inhibitory factor-kappa B (I-${\kappa}B$)의 활성을 촉진시킴으로써 nuclear factor-kappa B (NF-${\kappa}B$)의 인산화를 유도하였다. 마지막으로 비브리오균(WT)에 감염된 회장의 경우, 정상마우스에 비해 염증성 cytokine인 interleukin (IL)-6, IL-8, tumor necrosis factor (TNF)-${\alpha}$의 mRNA 수준이 유의적으로 증가되었다. 염증매개 수용체인 toll like receptor (TLR)-4, TLR-5, TLR-9의 mRNA의 발현 또한 비브리오균 처리에 의해 증가되어 있음이 관찰되었다. 종합적으로 오염된 식품 섭취를 통해 유입되는 비브리오균은 회장상피세포를 표적으로 염증반응을 일으키며, 그 기작은 PKC, ERK1/2, 그리고 JNK의 인산화를 통한 NF-${\kappa}B$ 활성의 촉진이며, 이로 인한 다양한 염증 매개 단백질 발현의 증가를 통해 이루어진다고 할 수 있다.