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Vibrio Vulnificus Induces the Inflammation of Mouse Ileal Epithelium: Involvement of Protein Kinase C and Nuclear Factor-Kappa B (회장 상피세포에서 비브리오균(Vibrio vulnificus)의 염증 유도 기작 연구: protein kinase C와 nuclear factor kappa-B의 관련성)

  • Han, Gi Yeon;Jung, Young Hyun;Jang, Kyung Ku;Choi, Sang Ho;Lee, Sei-Jung
    • Journal of Life Science
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    • v.24 no.6
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    • pp.664-670
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    • 2014
  • In the present study, we investigate the role of V. vulnificus in promoting the inflammation of mouse ileal ephitelium and its related signaling pathways. ICR mice were infected orally with V. vulnificus ($1{\times}10^9CFU$) for 16 h as a representative model of food-borne infection. To find the major portal of entry of V. vulnificus in mouse intestine, we have measured the levels of bacterial colonization in small intestine, colon, spleen, and liver. V. vulnificus appeared to colonize in intestine and colon in the order of ileum >> jejunum> colon, but lack in the duodenum, spleen, and liver. V. vulnificus in ileum caused severe necrotizing enteritis and showed shortened villi heights accompanied by an expanded width and inflammation, compared with the control mice. V. vulnificus induced ileal epithelium inflammation by activating phosphorylation of PKC and membrane translocation of $PKC{\alpha}$. V. vulnificus induced the phosphorylation of ERK and JNK, but did not affect p38 MAPK phosphorylation. Notably, V. vulnificus stimulated the I-${\kappa}B$-dependent phosphorylation of NF-${\kappa}B$ in mouse ileal epithelium. Finally, the ileal infection of V. vulnificus resulted in a significant increase in expression of proinflammatory cytokines and Toll-like receptors, respectively, compared to the control. Collectively, our results indicate that V. vulnificus induces ileal epithelium inflammation by increasing NF-${\kappa}B$ phosphorylation via activation of PKC, ERK, and JNK, which is critical for host defense mechanism in food-borne infection by V. vulnificus.

Anti-inflammatory effect of Polygonum multiflorum extraction in activated RAW 264.7 cells with lipopolysaccharide (Lipopolysaccharide로 활성화된 RAW 264.7 세포에서 적하수오(Polygonum multiflorum) 추출물의 항염증 효과 검증)

  • Lee, Eunsu;Kim, Hyeongjeong;Yu, Jae-Myo;Cho, Yong-Hun;Kim, Dong-In;Shin, Yuhyeon;Cho, Yeongje;Kwon, O-Jun;An, Bongjeon
    • Korean Journal of Food Preservation
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    • v.21 no.5
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    • pp.740-746
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    • 2014
  • The anti-inflammatory effects of Polygonum multiflorum water extracts (PMWs) and Polygonum multiflorum 70 % ethanol extracts(PMEs) were investigated using lipopolysaccharide-induce by inflammatory response. The inhibitory effects of PMWs and PMEs on the production of nitric oxide (NO) and pro - inflammatory cytokines in LPS - activated Raw 264.7 cells were investigated. The effects were examined after reducing production of Nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), interleukin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein levels. RAW 264.7 cells were cultured with LPS ($1{\mu}g/mL$) in the presence or absence of PMWs and PMEs for 24 h to determine their NO, iNOS, COX-2 levels. During the entire experimental period 10, 25, 50 and $100{\mu}g/mL$ of PMWs and PMEs showed no cytotoxicity. At these concentrations, PMWs and PMEs concentration dependently reduced the production of nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), interleukin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and interleukin-$1{\beta}$ (IL-$1{\beta}$). PMWs and PMEs were inhibited the activittion of iNOS, COX-2 by 89%, 54%, 91% and 57% respectively, at $100{\mu}g/mL$. These results indicate that PMWs and PMEs significantly reduces the effect of oxidative and inflammatory cytokines.

Medicinal Herb Extracts Attenuate 1-Chloro-2,4dinitrobenzene-induced Development of Atopic Dermatitis-like Skin Lesions (한약재 단일 추출물 및 복합 추출물을 이용한 아토피성 피부염 억제 효과)

  • Lee, Moon Hee;Han, Min Ho;Yoon, Jung Jeh;Song, Myung Kyu;Kim, Min Ju;Hong, Su Hyun;Choi, Byung Tae;Kim, Byung Woo;Hwang, Hye Jin;Choi, Yung Hyun
    • Journal of Life Science
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    • v.24 no.8
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    • pp.851-859
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    • 2014
  • The present study was designed to investigate whether ethanol extracts of Sophora flavescens (GS), Glycyrrhiza uralensis (GC), Dictamnus dasycarpus (BSP), and their mixtures (GGB-1, -2, -3, and -4) inhibit 1-chloro-2,4-dinitrobenzene (DNCB)-induced atopic dermatitis (AD) in a mouse model. DNCB was topically applied on the dorsal surface of Balb/c mice to induce AD-like skin lesions. The pathological phenotypes of AD, such as erythema, ear thickness, edema, scabs, and discharge, were significantly decreased in the GGB (DNCB + GS:GC:BSP = 3:1:1 mixture)-1-treated groups compared with the other treated groups. The weight of the spleen in immune organs was significantly decreased in the GGB-1-treated groups, whereas the weight of the liver in a control group was similar to that of the groups treated with the samples. Furthermore, toluidine blue staining analysis, a method used to specifically identify mast cells, showed that master cell infiltration into the dermis of the GGB-1-treated group was significantly decreased. The immunoglobulin E concentration was lower in the GGB-1-treated group. In addition, the levels of inflammatory cytokines (interferon-${\gamma}$, interleukin-1, 4, 5, 6, and 13, $1{\beta}$, and tumor necrosis factor-${\alpha}$) were also significantly reduced in the GGB-1-treated group. Taken together, these results suggest that a mixture of GS, GC, and BSP in a proportion of 3:1:1 (GGB-1) may contribute to the relief of AD symptoms and may be considered an excellent candidate for an AD therapeutic drug.

Anti-Inflammatory Effects of Extracts from Ligustrum ovalifolium H. Leaves on RAW264.7 Macrophages (RAW264.7 대식세포에서 왕쥐똥나무잎 추출물의 항염증 효과)

  • Kim, Yon-Suk;Lee, Seung-Jae;Hwang, Jin-Woo;Kim, Ee-Hwa;Park, Pyo-Jam;Jeong, Jae-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.9
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    • pp.1205-1210
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    • 2012
  • This study investigated the anti-inflammatory effects of Ligustrum ovalifolium H. (LOH) leaf extracts on RAW264.7 macrophages. Cell toxicity was determined by MTT assay. We evaluated the anti-inflammatory effects of LOH extracts by measuring nitric oxide (NO), reactive oxygen species (ROS), inducible NOS (iNOS) production, and cyclooxygenase-2 (COX-2) expression by Western blotting. LOH ethanolic extracts (0.05, 0.1, and 0.2 mg/mL) significantly suppressed LPS-stimulated production of NO. The intracellular ROS level also significantly decreased. LOH ethanolic extracts reduced the expression of iNOS and COX-2 proteins. The present results show that LOH ethanol extract has potent anti-inflammatory effects on RAW264.7 macrophages. These results also suggest that the anti-inflammatory effects of LOH extracts may be related to the inhibition of LPS-stimulated ROS and NO production. Therefore, ethanolic extracts of LOH leaves may be utilized as a good source of functional foods for protection against inflammatory diseases.

The Effect of Samul-tanggahyangbuja on Anti-Depressive Behavior and Immunity (사물탕가향부자가 항우울행동 및 면역기능에 미치는 영향)

  • Jang, Yoon-Jeong;Kim, Song-Baek;Choi, Chang-Min;Seo, Yun-Jung;Cho, Han-Baek
    • The Journal of Korean Obstetrics and Gynecology
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    • v.26 no.4
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    • pp.14-29
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    • 2013
  • Objectives: Samul-tang and Rhizome of Cyperus rotundus L. have been frequently used in gynecologic disease. The purpose of the present study is to explore the behavioral and neurobiological effects of Samul-tanggahyangbuja (SGH) on ovariectomized rats and to form a basis for clinical treatment. Methods: Ovariectomized rats were repeatedly stressed for over 2 weeks. After orally medicated with SGH (200 or 400 mg/kg/day), the anxiety response was tested using the Elevated Plus Maze (EPM) in rats. The serum levels of estradiol and IL-4, and immunohistochemical changes of IL-4 in the Locus coeruleus (LC) and Paraventricular Nucleus (PVN) were measured. Results: 1. In the EPM, SGH 400 mg significantly increased time spent on the open arms and decreased time spent on the closed arms, compared with the control group (p<0.05). 2. SGH tended to increase numbers of crossings in the open and closed arms in the EPM. However, it did not reach statistical significance. 3. SGH significantly increased the serum levels of estradiol compared with the control group (p<0.05). 4. SGH 400 mg significantly increased the serum levels of IL-4 compared with the control group (p<0.05). 5. IL-4 immunoreactivity was reduced in the control group compared with the normal group (p<0.05). However, SGH groups (200 and 400 mg) did not produce any significant effects on levels of IL-4 in the LC and PVN. Conclusions: These results suggest that SGH possesses the anti-depressant and immuno-modulatory effects on ovariectomized rats.

Anti-inflammatory Effect of Flavonoids Kaempferol and Biochanin A-enriched Extract of Barnyard Millet (Echinochloa crus-galli var. frumentacea) Grains in LPS-stimulated RAW264.7 Cells (마우스 대식 세포주 RAW264.7에 있어서 LPS처리에 의해 유도되는 염증반응에 대한 식용피(Echinochloa crus-galli var. frumentacea)의 저해효과)

  • Lee, Ji Young;Jun, Do Youn;Yoon, Young Ho;Ko, Jee Youn;Woo, Koan Sik;Woo, Mi Hee;Kim, Young Ho
    • Journal of Life Science
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    • v.24 no.11
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    • pp.1157-1167
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    • 2014
  • In order to compare the anti-inflammatory effects of five selected cereal grains-proso millet, hwanggeumchal sorghum, foxtail millet, barnyard millet, and adlay-the inhibitory activities of 80% ethanol (EtOH) extracts obtained from the individual grains on lipopolysaccharide (LPS)-induced nitric oxide (NO) generation were investigated in RAW264.7 cells. The EtOH extract of barnyard millet (Echinochloa crus-galli var. frumentacea) grains exhibited more potent anti-inflammatory activity than that of the other grains. When the EtOH extract of barnyard millet grains was sequentially fractionated with n-hexane, methylene chloride (MC), ethyl acetate (EtOAc), and n-butanol, the majority of the anti-inflammatory activity was detected in the MC fraction, followed by the EtOAc fraction. Pretreatment with the MC fraction caused downregulation of the expression levels of iNOS- and COX-2-specific transcripts and proteins, as well as proinflammatory cytokine gene transcripts (IL-$1{\beta}$, IL-6, and TNF-${\alpha}$) in LPS-stimulated RAW264.7 cells. Additionally, the MC fraction could suppress not only the LPS-induced nuclear translocation of cytosolic NF-kB, but also the LPS-induced activation of MAPKs, such as ERK, JNK, and p38MAPK. Further analysis of the MC fraction by HPLC identified kaempferol, biochanin A, and formononetin as the major phenolic components. Both kaempferol and biochanin A, but not formononetin, could exert anti-inflammatory effect at the same concentrations as those of the MC fraction. Consequently, these results indicate that kaempferol and biochanin A are among the most effective anti-inflammatory phenolic components in barnyard millet grains. This finding suggests that barnyard millet grains and the MC extract enriched in kaempferol and biochanin A could be beneficial functional food sources that have an anti-inflammatory effect.

Anti-inflammatory Effect of Lactuca sativa L. Extract in Human Umbilical Vein Endothelial Cells and Improvement of Lipid Levels in Mice Fed a High-fat Diet (상추 추출물(Lactuca sativa L.)의 혈관내피세포에서 항염증 작용과 고지방 식이 생쥐에서 혈중 지질농도 개선에 미치는 영향)

  • Hwang-Bo, Jeon;Jang, Kyung Ok;Chung, Hayoung;Park, Jong-Hwa;Lee, Tae Hoon;Kim, Jiyoung;Chung, In Sik
    • The Korean Journal of Food And Nutrition
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    • v.29 no.6
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    • pp.998-1007
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    • 2016
  • The objective of this study was to investigate the effects of a lettuce (Lactuca sativa L.) extract on the inflammation of human umbilical vein endothelial cell (HUVEC) and blood lipid improvement in hypercholesterolemic mice fed a high cholesterol diet. The lettuce extract (100% ethanol extract) inhibited the expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 in HUVEC treated with tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$). The lettuce extract suppressed the adhesion of THP-1 to TNF-${\alpha}$-treated HUVEC. The lettuce extract decreased the TNF-${\alpha}$-stimulated production of proinflammatory cytokine interleukin-6, interleukin-8 and chemokine monocyte chemotactic protein 1. In hypercholesterolemic mice, the lettuce extract reduced serum total cholesterol, triglyceride, and low-density lipoprotein-cholesterol level, while the lettuce extract elevated high-density lipoprotein-cholesterol level, resulting in the decrease of atherogenic index and cardiac risk factor level. These results suggested that lettuce extract can be an useful resource to show an anti-inflammatory effect and improve lipid metabolism.

Anti-Inflammatory Effect of Ethanol Extract from Grateloupia crispata on Lipopolysaccharide-Induced Inflammatory Responses in RAW 264.7 Cells and Mice Ears (LPS로 유도된 RAW 264.7 세포와 마우스 귀 조직에 대한 주름까막살 에탄올 추출물의 항염증 효과)

  • Bae, Nan-Young;Kim, Min-Ji;Kim, Koth-Bong Woo-Ri;Park, Sun-Hee;Jang, Mi-Ran;Ahn, Dong-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.8
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    • pp.1090-1098
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    • 2016
  • The anti-inflammatory effects of ethanol extract from Grateloupia crispata (GCEE) were investigated in lipopolysaccharide (LPS)-stimulated murine macrophages. Anti-inflammatory effects were detected by enzyme-linked immunosorbent assay, Western blotting, and immunohistochemistry. There was no cytotoxic effect on proliferation of macrophages treated with GCEE compared to the control. GCEE significantly inhibited production of pro-inflammatory cytokines [interleukin (IL)-6, tumor necrosis $factor-{\alpha}$, and $IL-1{\beta}$] as well as nitric oxide in LPS-stimulated RAW 264.7 cells. In addition, GCEE suppressed expression of inducible nitric oxide synthase, cyclooxygenase-2, and nuclear $factor-{\kappa}B$ in a dose-dependent manner. GCEE significantly reduced activation of mitogen-activated protein kinases. In the in vivo test, evaluation of anti-inflammatory activity of GCEE was performed using croton oil-induced ear edema in ICR mice. Oral administration of 10 mg/kg to 250 mg/kg of GCEE significantly reduced ear edema in a dose-dependent manner compared to croton oil-induced mice. Moreover, GCEE reduced ear thickness and the number of mast cells compared to croton oil-induced mice in the histological analysis. These data suggest that GCEE could be used as a potential source for anti-inflammatory agents.

Anti-inflammatory Effect of Myricetin from Rhododendron mucronulatum Turcz. Flowers in Lipopolysaccharide-stimulated Raw 264.7 Cells (Lipopolysaccharide로 유도된 Raw264.7 cell에서 Rhododendron mucronulatum Turcz. Flower으로부터 분리한 myricetin에 의한 염증 억제효과)

  • Choi, Moo-Young;Hong, Shin-Hyup;Cho, Jun-Hyo;Park, Hye-Jin;Jo, Jae-Bum;Lee, Jae-Eun;Kim, Dong-Hee;Kim, Byung-Oh;Cho, Young-Je
    • Journal of Life Science
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    • v.26 no.11
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    • pp.1245-1252
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    • 2016
  • As a research of inflammation inhibitory activity using natural resource, the inflammation inhibitory activity by purified active compound from Rhododendron mucronulatum flower was experimented. Rhododendron mucronulatum flower components were purified and separated with Sephadex LH-20 and MCI gel CHP-20 column chromatography, Purified compound was confirmed as myricetin by $^1H-NMR$, $^{13}C-NMR$ and Fast atom bombardment (FAB)-Mass spectrum to have inhibition activity on inflammatory factors secreted by Raw 264.7 cells in response to lipopolysaccharide stimulation. Myricetin inhibited nitric oxide (NO) expression in a concentration dependent manner, approximately 40% inhibition was observed at a concentration of $50{\mu}M$. The inhibition effect of myricetin on inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 protein expression was 20% and 80%, respectively, at a concentration of $25{\mu}M$. Myricetin also inhibited expression of the inflammatory cytokines, tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\beta}$, IL-6 and prostaglandin $E_2(PGE_2)$ in a concentration dependent manner; a concentration of $50{\mu}M$, 70%, 80%, 80% and 95% inhibition was observed, respectively. Therefore myricetin isolated from Rhododendron mucronulatum flowers is expected to have an anti-inflammatory effect in Raw 264.7 cell induced by lipopolysaccharides. The results can be expected myricetin from Rhododendron mucronulatum flower to use as functional resource for anti-inflammatory activity.

Anti-Inflammatory Activity of Ethanol Extracts from Hizikia fusiformis Fermented with Lactic Acid Bacteria in LPS-Stimulated RAW264.7 Macrophages (유산균 종류에 따른 발효톳 추출물의 항염증 활성)

  • Kwon, Myeong Sook;Mun, Ok-Ju;Bae, Min Joo;Lee, Seul-Gi;Kim, Mihyang;Lee, Sang-Hyeon;Yu, Ki Hwan;Kim, Yuck Yong;Kong, Chang-Suk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.10
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    • pp.1450-1457
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    • 2015
  • The anti-inflammatory effect of ethanol extracts from Hizikia fusiformis fermented with and without lactic acid bacteria was compared in lipopolysaccharide (LPS)-stimulated RAW 264.7 mouse macrophages. The fermentation was done using Weissella sp. SH-1 and Lactobacillus casei in a mixture of glucose and lactate source at $30^{\circ}C$ for 30 days. As a result, we confirmed that the fermentation of H. fusiformis with lactic acid bacteria inhibited LPS-stimulated nitric oxide (NO) production and the expression of inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2, interleukin (IL)-6, tumor necrosis factor ${\alpha}$, and IL-$1{\beta}$ as important inflammatory factors. During a comparison analysis, we found that L. casei fermented groups significantly suppressed NO production by regulating iNOS and COX-2 expression. Also, the effective suppression of pro-inflammatory cytokine and LPS-induced activation of mitogen- activated protein kinase indicated that the fermentation using Weissella sp. SH-1 and L. casei may provide an increment towards the extraction of active components, which are effective anti-inflammatory agents.