• Title/Summary/Keyword: Nitric Oxide(No)

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Role of NO in Activation of $NF{\kappa}B$ by PM2.5 in Lung Epithelial Cells (PM2.5로 자극한 폐상피세포의 $NF{\kappa}B$ 활성화에 NO의 역할)

  • Kim, Kyoung-Ah;Nam, Hae-Yun;Mun, Je-Hyeok;Jeong, Jin-Sook;Lim, Young;Kai, Hirofumi
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.6
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    • pp.616-626
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    • 2002
  • Background : The present study was performed to further improve our understanding of molecular mechanisms involved in the activation of NFkB, a major transcriptional factor involved in the inflammatory response in the lung, by particulate matter in lung epithelial cells with an aerodynamic diameter of less than $2.5{\mu}m$(PM2.5). Materials and Methods : Immediate production of reactive oxygen species (ROS) and nitrogen species (RNS), with the PM2.5 induced expression of inducible nitric oxide synthase (iNOS), $I{\kappa}B$ degradation and $NF{\kappa}B$-dependent transcriptional activity, in 549 cells, were monitored. Addition, we also examined the effect of the iNOS inhibitor, L-N6-(1-iminoethyl) lysine hydrochloride (L-NIL), on the PM2.5-induced $NF{\kappa}B$ activation in A549 cells. Results : The rapid degradation of $I{\kappa}B$ and the increase of transcriptional activity of the $NF{\kappa}B$-dependent promotor were observed in A549 cells exposed to PM2.5. The immediate production of ROS in response to PM2.5 in A549 cells was not clearly detected, although immediate responses were observed in RAW264.7 cells. A 549 cells, cultured in the presence of PM2.5, produced an increase in NO, which was noticeably significant after 15 min of exposure with the expression of iNOS mRNA. The addition of L-NIL, an iNOS inhibitor, significantly inhibited the PM2.5-induced $I{\kappa}B$ degradation and the increase of the $NF{\kappa}B$-dependent transcriptional activity. Conclusion : These results suggest that PM2.5 stimulates the immediate production of RNS, leading to the activation of $NF{\kappa}B$ in the pulmonary epithelium.

Anti-inflammatory Activity of Extracts of Hovenia dulcis on Lipopolysaccharides-stimulated RAW264.7 Cells (LPS로 유도된 RAW264.7 대식세포에 대한 헛개나무(Hovenia dulcis) 추출물의 항염증 효과)

  • Woo, Hyun Sim;Lee, Sun Min;Heo, Jeong Doo;Lee, Min-Sung;Kim, Yeong-Su;Kim, Dae Wook
    • Korean Journal of Plant Resources
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    • v.31 no.5
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    • pp.466-477
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    • 2018
  • In this study, the anti-inflammatory activities of the extracts of different parts of Hovenia dulcis such as leaves, stems, and roots were investigated. Among them, the roots extract (RE) showed the most potent suppressive effect against pro-inflammatory mediators in LPS-stimulated mouse macrophage cells. RE induced dose-dependent reduction of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) and concomitantly reduced the production of NO and $PGE_2$. Additionally, pre-treatment with RE significantly suppressed the production of inflammatory cytokines, such as tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), interleukin $(IL)-1{\beta}$, and IL-6, as well as mRNA levels. Moreover, phosphorylation of mitogen-activated protein kinases (MAPKs) and nuclear translocation of nuclear factor-kappa B (NF-kB) were also strongly attenuated by RE in RAW264.7 cell. Furthermore, RE induced HO-1 expression through nuclear translocation of nuclear factor E2-related factor 2 (Nrf2) and increase HO-1 activity in RAW264.7 macrophages. Therefore, these results indicate that RE strongly inhibits LPS-induced inflammatory responses by blocking NF-kB activation, inhibiting MAPKs phosphorylation, and enhancing HO-1 expression in macrophages, suggesting that RE of H. dulicis and a major component, 27-O-protocatechuoylbetulinic acid could be applied as a valuable natural anti-inflammatory material.

Enhancement of Immune Activities of Peptides from Asterias amurensis Using a Nano-encapsulation Process (나노 입자 불가사리 펩타이드의 면역 활성 증진)

  • Jeong, Hyang-Suk;Oh, Sung-Ho;Kim, Seoung-Seop;Jeong, Myoung-Hoon;Choi, Woon-Yong;Seo, Yong-Chang;Choi, Geun-Pyo;Kim, Jin-Chul;Lee, Hyeon-Yong
    • Korean Journal of Food Science and Technology
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    • v.42 no.4
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    • pp.424-430
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    • 2010
  • Immuno-modulatory activities of peptides from Asterias amurensis were investigated using a nano-encapsulation process. The molecular weights of the peptides in the range of 5-7 kDa were separated using Sephadex G-75 gel filtration. Eighty-five percent of the nano-particles were in the 300 nm range using dynamic light scattering. The cytotoxicity of the A. amurensis nano-particles against CCD-986sk human dermal fibroblast cells was 11.64% after adding 1.0 mg/mL of the samples, which was lower than that from the control (13.28% collagen). The secretion of $NO^-$ from macrophages was estimated as $40\;{\mu}M$ after adding 1.0 mg/mL of gelatin nano-particles, which was higher than the others. Prostaglandin $E_2$ production from UV-induced human skin cells decreased greatly to 860 pg/mL after adding 1.0 mg/mL of the samples. Confocal microscopy revealed that nano-particles effectively penetrated the cells within 1 hour. From these results, we consider that nano-encapsulation of the peptides from A. amurensis can improve their biological functions.

Enhancement of Skin Immune Activation Effect of Collagen Peptides Isolated from Asterias amurensis (불가사리 유래 콜라겐 펩타이드의 피부 면역 증진 효과)

  • Jeong, Hyang-Suk;Kwon, Min-Chul;Han, Jae-Gun;Ha, Ji-Hye;Jin, Ling;Kim, Jin-Chul;Kwak, Hyeong-Geun;Hwang, Bo-Young;Lee, Hyeon-Yong
    • Korean Journal of Food Science and Technology
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    • v.40 no.5
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    • pp.522-527
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    • 2008
  • Low molecular peptides were isolated from Asterias amurensis via SDS-PAGE. The peptides were separated via consecutive gel filtration as five fractions (F1-F5) according to molecular weights, based on the results of MALDI-TOF MS analysis. The molecular weight of the most active peptide was estimated as 15,000 daltons. The peptide showed cytotoxicity on normal human fibroblast cells at levels as low as 20% when 1.0 mg/mL of the samples was added. The peptide also exhibited higher levels of nitric oxide production from macrophages than the lipopolysaccaharides. It was determined that prostaglendin $E_2$ production was significantly inhibited, up to 127.8% as compared to the control. The low molecular peptide inhibited hyaluronidase activity as 535.7 ${\mu}g/mL$ of $IC_{50}$. It can be concluded that the relatively low molecular weight peptide, fucoidan, from A. amurensis has excellent cosmetic and immunomodulatory activities, which can be considered as a possible resource of new cosmetic agents for skin immunomodulation.

INFLUENCES OF APICOECTOMY AND RETROGRADE CAVITY PREPARATION METHODS ON THE APICAL LEAKAGE (치근단절제 및 역충전와동 형성방법이 치근단누출에 미치는 영향)

  • Yang, Jeong-Ok;Kim, Sung-Kyo;Kwon, Tae-Kyung
    • Restorative Dentistry and Endodontics
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    • v.23 no.2
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    • pp.537-549
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    • 1998
  • The purpose of this study was to evaluate the influence of root resection and retrograde cavity preparation methods on the apical leakage in endodontic surgery. To investigate the effect of various root resection and retrograde cavity preparation methods on the apical leakage, 71 roots of extracted human maxillary anterior teeth and 44 mesiobuccal roots of extracted human maxillary first molars were used. Root canals of the all the specimens were prepared with step-back technique and filled with gutta-percha by lateral condensation method. Three millimeters of each root was resected at a 45 degree angle or perpendicular to the long axis of the tooth according to the groups. Retrograde cavities were prepared with ultrasonic instruments or a slow-speed round bur, and occlusal access cavities were filled with zinc oxide eugenol cement. Three coats of clear nail polish were placed on the lateral and coronal surfaces of the specimens except the apical cut one millimeter. All the specimens were immerged in 2% methylene blue solution for 7 days in an incubator at $37^{\circ}C$. The teeth were dissolved in 14 ml of 35% nitric acid solution and the dye present within the root canal system was returned to solution. The leakage of dye was quantitatively measured via spectrophotometric method. The obtained data were analysed statistically using two-way ANOVA and Duncans Multiple Range Test. The results were as follows: 1. No statistically significant difference was observed between ultrasonic retrograde cavity preparation method and slow-speed round bur technique, without apical bevel (p>0.05). 2. Ultrasonic retrograde preparation method showed significantly less apical leakage than slow-speed round bur technique, with bevel (p<0.0001). 3. No statistically significant difference was found between beveled resected root surface and non-beveled resected root surface, with ultrasonic technique (p>0.05). 4. Non-beveled resected root surface showed significantly less apical leakage than beveled resected root surface, with slow-speed round bur technique (p<0.0001). 5. No statistically significant difference in apical leakage was found between the group of retrograde cavity prepared parallel to the long axis of the tooth and the group of one prepared perpendicular to the long axis of the tooth (p>0.05). 6. Regarding isthmus preparation, ultrasonic retrograde preparation method showed significantly less apical leakage than slow-speed round bur technique, in the mesiobuccal root of maxillary molar, without bevel (p<0.0001).

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Anti-inflammatory Effects of Rebaudioside A in LPS-stimulated RAW264.7 Macrophage Cells (LPS에 의해 자극된 RAW264.7 대식세포에서 Rebaudioside A의 항염 효과)

  • Cho, Uk Min;Hwang, Hyung Seo
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.43 no.2
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    • pp.157-164
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    • 2017
  • Stevia (Stevia rebaudiana) is a perennial plant of the genus Stevia, originated in South America. It stores many forms of glycosides, mainly stevioside and rebaudioside A, in which steviol is the basic structure. Steviol glycosides, widely used as sweeteners, are superior to sugar in sweetness. Recently, it has been reported that steviol glycosides are involved not only in the skin whitening and anti-inflammatory effect but also in enhancing skin barrier function through tight junction regulation. Thus, we examined anti-inflammatory effect of rebaudioside A and tried to identify its potential for improving atopic dermatitis as cosmetic ingredients. To investigate the anti-inflammatory effect, cell viability and mRNA expression level of inflammation-related cytokines were measured using mouse macrophage RAW264.7 cells. Cell counting kit 8 (CCK-8) assay was carried out to measure cell viability and the maximum concentration without cytotoxicity was set to $250{\mu}M$. A quantitative real-time RT-PCR method was used for the study of the inflammatory suppression of rebaudioside A. Rebaudioside A inhibited expression of inducible nitric oxide synthase (iNOS) up to 47% and COX-2 up to 41% compared to LPS treated condition. NO synthesis was decreased by rebaudioside A. Also, mRNA expression of interleukin (IL)-$1{\alpha}$, IL-$1{\beta}$ and IL-6 in LPS-stimulated RAW264.7 cells was decreased to 40%, 45% and 59%, respectively, as a concentration-dependent manner. In conclusion, rebaudioside A inhibited the inflammatory response by regulation of cytokine gene expression. From these results, we expect that steviol glycoside, such as rebaudioside A, can be used as a material for improving atopic dermatitis in the future.

Studies on the Comparative Analysis of Immunofunction of Agaricus blazei Murill Cultivated with Fermented Media Containing Pueraria thunbergiana (칡혼합 발효배지로 생산된 신령버섯의 면역기능성 비교 분석에 관한 연구)

  • 김주남;서정식;박동철
    • Food Science and Preservation
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    • v.9 no.1
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    • pp.114-119
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    • 2002
  • This research was performed to investigate the immnomodulative effects of ploysaccharides extracted from the fruiting body of Agarcus blazei cultivated with the media which are fermented with sugar cane bagasse containing Pueraria thunbergiana in open-air storage. In MTT test, methanol extracts from the fruiting body of A. blazei cultivated with P. thunbergiana media showed in colon carcinoma line(HT29) by 1.5∼3.5 fold and human heptoma cell line (HepG2) by 1.3 ∼2.4 fold antitumor activites compared to two types media (rice straw plus sugar cane bagasse, rice straw only) often used in the fauns. To clarify the antimutagenic principles, three extracts, Ab-l, Ab-2 and Ab-3, were separated by the solvent fractionations such as hot water, cold & hot sodium hydroxide respectively, and their antimutagenic effects was determined against N-methyl-N'-nitro-N-cnitrso-guanidine(MNNG) using Salmonella typhymurium. There was no significant differencies of inhibition levels among the used media, but Ab-3 tractions still showed a high antimutagenicity in the Ames test regardless of cultivating areas or media. To prove the cell immunofunction, nitric oxide (NO) produced from Raw 264.7 matrophage cultured with three fractions (Ab-l, Ab-2, Ab-3) was measured, and showed generally increase about 45 ∼58 percent compared to another two media (rice straw plus sugar cane bagasse, rice straw only), in the fraction of hot alklai extracts of the fruiting body cultivated with P. thunbergiana, which means that the media selection could be very important factors for improving medicinal effects in agaricus blazei fruiting body.

Sargassum sp. Attenuates Oxidative Stress and Suppresses Lipid Accumulation in vitro (모자반추출물의 항산화활성 및 지방세포 생성억제 효과)

  • Kim, Jung-Ae;Karadeniz, Fatih;Ahn, Byul-Nim;Kwon, Myeong Sook;Mun, Ok-Ju;Kim, Mihyang;Lee, Sang-Hyeon;Yu, Ki Hwan;Kim, Yuck Yong;Kong, Chang-Suk
    • Journal of Life Science
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    • v.24 no.3
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    • pp.274-283
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    • 2014
  • Oxidative stress causes tissue damage and facilitates the progression of metabolic diseases, including diabetes, cardiovascular heart diseases, and obesity. Lipid accumulation and obesity-related complications have been observed in the presence of extensive oxidative stress. As part of an ongoing study to develop therapeutic supplements, Sargassum sp. were tested for their ability to scavenge free radicals and intracellular reactive oxygen species (ROS), as well as to suppress lipid accumulation. Three species, S. hemiphyllum, S. thunbergii, and Sargassum horneri, were shown to scavenge free radicals in a di(phenyl)-(2,4,6-trinitrophenyl)iminoazanium (DPPH) assay. In addition, Sargassum sp. was shown to scavenge intracellular ROS and to decrease nitric oxide (NO) production in $H_2O_2$ and lipopolysaccharide (LPS)-induced in RAW264.7 mouse macrophages, respectively. Taken together, the results suggest that Sargassum sp. possess huge potential to relieve oxidative stress and related complications, as well as lipid-induced oxidation. They indicate that S. hemiphyllum, S. thunbergii, and S. horneri are potent functional supplements that can produce beneficial health effects through antioxidant and antiobesity activities, with S. hemiphyllum being the most potent among the Sargassum sp. tested. A potential mechanism for the effect of Sargassum sp. on the suppression of lipid accumulation in differentiating 3T3-L1 mouse preadipocytes through deactivation of the peroxisome proliferator-activated receptor ${\gamma}$ (PPAR ${\gamma}$) is presented.

Anti-Inflammatory Effects of Volatile Flavor Extract from Herbal Medicinal Prescriptions Including Cnidium officinale Makino and Angelica gigas Nakai (천궁 및 당귀를 함유한 한방처방제 휘발성 향기추출물의 항염증 효과)

  • Leem, Hyun-Hee;Kim, Eun-Ok;Seo, Mi-Jae;Choi, Sang-Won
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.37 no.3
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    • pp.199-210
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    • 2011
  • This study was conducted to develop functional sources of herbal cosmetics for treatment of skin aging and inflammatory disorders using volatile flavor extracts of four different herbal medicinal prescriptions including Cnidium officinale Makino (COM), Angelica gigas Nakai (AGN), Mentha arvense L. (MAL), Artemisiae argyi Folium (AAF), Paeonia lactiflora Pall (PLP), Rehmanniae Radix Preparata (RRP), Scutellaria baicalensis Georgi (SBG), Panax ginseng C.A. Meyer (PGM), Glycyrrhiza uralensis Fisch (GUF). The volatile flavor extracts of four different herbal medicinal prescriptions (HH-1: COM, AGN, PLP, RRP, HH-2: COM, AGN, PLP, RRP, SBG, PGM, GUF, HH-3: COM, AGN, MAL, AAF, HH-4: COM, AGN, MAL, AAF, SBG, PGM, GUF) were extracted using SDE and their antioxidant and anti-inflammatory effects were measured by using DPPH radical and SLO, respectively. As a result, HH-2 showed moderate DPPH radical scavenging activity (68.24 %) and the strongest SLO inhibitory activity (83.96 %) at 100 ${\mu}g$/mL. Moreover, HH-2 of four different prescriptions significantly inhibited NO production on LPS-stimulated RAW 264.7 cells in a dose-dependent manner without considerable cell cytotoxicity at range of 2.0 ~ 50 ${\mu}g$/mL. Additionally, HH-2 also effectively suppressed the production of $PGE_2$ and IL-6, which are responsible for promoting the inflammatory process. Major volatile components of HH-2 were identified as eugenol, paeonol, butyl phthalide, ${\beta}$-eudesmol and butylidene dihydrophthalide by GC-MS analysis. Thus, these results suggest that HH-2 may be useful as a potential source of anti-inflammatory agents in herbal medicinal cosmetics.

Anti-inflammatory and Antioxidant Effects of Spiraea prunifolia Sieb. et Zucc. var. simpliciflora Nakai in RAW 264.7 Cells (조팝나무 뿌리 열수 추출물이 RAW264.7 세포에서 미치는 항산화 및 항염증 활성)

  • Sim, Mi-Ok;Lee, Hyun Joo;Jang, Ji Hun;Lee, Hyo Eun;Jung, Ho-Kyung;Kim, Tae-Muk;No, Jong hyun;Jung, Jakyun;Jung, Da Eun;Cho, Hyun-Woo
    • Korean Journal of Plant Resources
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    • v.30 no.4
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    • pp.335-342
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    • 2017
  • Spiraea prunifolia Sieb. et Zucc. var. simpliciflora Nakai (SSN) has been used for the anti-inflammation in traditional folk medicine. To compare water and methanol extracts of SSN, we analyzed major components using LC IT TOF MS. The major components of hot water extract were identified as caffeic acid and p-coumaric acid, but methanol extract was not well established. However, methanol extract was detected with less polarity compounds compared to hot water extract. Next, we investigated the inhibitory effects of SSN water extract on the lipopolysaccharide (LPS)-induced inflammatory response or $H_2O_2-induced$ oxidative stress in Raw 264.7 macrophage cells. SSN strongly suppressed the production of nitric oxide in LPS-induced inflammatory response without cytotoxcity. The SSN possessed free radical scavenging activities such as DPPH ($IC_{50}=320.2{\mu}g/m{\ell}$), ABTS ($IC_{50}=124.0{\mu}g/m{\ell}$), and superoxide anion radical ($IC_{50}=122.6{\mu}g/m{\ell}$). The total phenol and flavonoid content of SSN was 56.7 mg/g, and 15.1 mg/g, respectively. Furthermore, SSN decreased the $H_2O_2-induced$ cytotoxicity by enhancing the cell viability, and SSN significantly reduced the intracellular reactive oxygen species (ROS) level. Therefore, SSN may be recommended as an effective strategy to prevent and/or treat various inflammation and ROS-induced diseases.