• Title/Summary/Keyword: Anti-inflammatory Effect

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Antioxidative and Antiallergic Effect of Persimmon Leaf Extracts (감잎(Diospyros kaki Thunb) 추출물의 항산화 및 항알레르기 효과)

  • Yoo, Ki-Hwan;Jeong, Jong-Moon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.12
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    • pp.1691-1698
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    • 2009
  • The purpose of this study was to investigate the antioxidative and antiallergic effects of persimmon leaf extract. Antioxidative and anti-inflammatory effects of the crude persimmon leaf extract (PLE) and the partially purified persimmon leaf extract (PPLE) were determined in in vitro assays by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide anion radicals, and 5-lipoxygenase (5-LO) and cyclooxygenase (COX). Total phenols and total flavonoid levels of PLE and PPLE were $230.0{\pm}19.6$ mg/g and $475.5{\pm}38.7$ mg/g, and $34.8{\pm}6.5$ mg/g and $78.8{\pm}3.6$ mg/g, respectively. DPPH and superoxide radical-scavenging activities ($SC_{50}$) of the PLE and PPLE were $23.8{\pm}3.2$ ppm and $10.0{\pm}1.3$ ppm, and $47.6{\pm}3.4$ ppm and $22.4{\pm}3.3$ ppm, respectively. Inhibitory activities ($IC_{50}$) of PLE and PPLE against 5-LO, COX-1 and COX-2 were $77.1{\pm}11.7$, $38.6{\pm}7.0$ ppm, $47.4{\pm}7.7$, $25.3{\pm}6.3$ ppm, and $129.5{\pm}5.5$, $84.5{\pm}2.3$ ppm, respectively. Moreover, two extracts inhibited dose-dependently NO production in LPS-stimulated RAW 264.7 cells, and also effectively inhibited the cutaneous anaphylaxis reaction activated by anti-dinitrophenyl IgE antibody in mice. These results suggest that PLE and PPLE may be useful for phytochemical materials for prevention and treatment of radical-mediated pathological and allergy diseases.

Physiological Properties of Jeju Traditional Doenjang (제주 재래식된장의 생리적 특성)

  • Hwang, Joon-Ho;Oh, You-Sung;Lim, Ja-Hun;Park, Ji-Eun;Kim, Mi-Bo;Yoon, Hoon-Seok;Lim, Sang-Bin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.12
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    • pp.1656-1663
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    • 2009
  • The antioxidant activities of water extracts from wild vegetables such as Ligularia fischeri (GC), Capsicum annuum L. (GCY), Aster scaber (CNM), Petasites japonicus S. et Z. Max (MYD), Ipomoea batatas L. (Lam) (GGM) were evaluated and compared with water extracts from freeze dried block. The antioxidant properties of water extracts from wild vegetables and their freeze dried block were evaluated using different antioxidant tests; 2.2-diphenyl-1-picryl-hydrazyl (DPPH) radical scavenging, hydroxyl radical scavenging and nitrite scavenging activities. The water extracts from wild vegetables were found to have a higher total phenolic content than water extracts from freeze dried block. Total phenolic contents of water extracts from GC, GCY, CNM, MYD, and GGM were $471.66{\pm}3.52\;{\mu}g/mg,\;141.33{\pm}2.51\;{\mu}g/mg,\;177.33{\pm}2.88\;{\mu}g/mg,\;238.66{\pm}9.50\;{\mu}g/mg\;and\;122.67{\pm}3.51\;{\mu}g/mg$, respectively. At the concentrations of 1000 ppm, water extracts from GC, GCY, CNM, and GGM showed higher activities than water extracts from their freeze dried block on DPPH radical scavenger activity. The activity of water extracts from CNM, GC, GCY, MYD, and GGM were 90.9%, 89.9%, 76.6%, 71.1%, and 57.4%, respectively. When 10000 ppm of GC, GCY, CNM, MYD, and GGM water extracts tested for hydroxyl radical scavenging activity, activities were increased by 38.8%, 33.4%, 35.9%, 34.3%, and 33.8%, respectively and a similar effect was found with water extracts from GCY, CNM, and GGM freeze dried block at 10000 ppm concentration. However, the water extracts from GC and MYD was slightly more effective than freeze dried block extracts. The water extracts from wild vegetables and their freeze dried block had effective DPPH radical scavenger activity and hydroxyl radical scavenging activity at all tested concentrations. Nitrite scavenging activity of GC water extract significantly increased in a dose-dependent manner and the extract had higher nitrite scavenging activity than extracts freeze dried block extracts. We found that freeze dried block maintained antioxidant activities of the wild vegetables.

Study on the Antioxidant and Human Neutrophil Elastase Inhibitory Activities of Mushroom Ramaria formosa Extracts (붉은싸리버섯 추출물의 항산화 및 Human Neutrophil Elastase 저해활성)

  • Kim, Kwan-Chul;Kwon, Yong-Beom;Jang, Hae-Dong;Kim, Jae Wha;Jeong, Jae Cheol;Lee, Ik-Soo;Ha, Byung-Jo;Yoo, Ick-Dong
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.42 no.3
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    • pp.269-278
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    • 2016
  • In searching for novel agents for skin anti-aging from natural resources, we found that the extract of the fruiting bodies of Ramaria formosa (R. formosa) had significant antioxidant and human neutrophil elastase (HNE) inhibitory activities. R. formosa extract exhibited a considerable DPPH radical scavenging activity with an antioxidant content of 117.0mg/mL (ascorbic acid equivalents) at the concentration of $500{\mu}g/mL$. The capacity of R. formosa extract to scavenge peroxy radicals measured by ORAC assay also showed dose-dependent antioxidant effect with $ORAC_{Roo}$ (trolox equivalents, $1{\mu}M$) values of 0.8, 5.2, and 7.8 at the concentrations of 1, 10, and $20{\mu}g/mL$. The cellular antioxidant capacity of R. formosa extract was investigated by assaying the cellular fluorescence intensity using dichlorodihydrofluorescein (DCF). The cellular oxidative stress induced by AAPH, $Cu^{2+}$ or $H_2O_2$ in HepG2 cells was significantly attenuated by more than 30% at $20{\mu}g/mL$ of R. formosa extract. HNE activity was reduced by treatment with R. formosa extract in a dose-dependent manner, and the $ED_{50}$ value for the ethanol extract of R. formosa was $42.9{\mu}g/mL$. R. formosa extract did not exhibited antimicrobial activity against four microorganisms including Bacillus subtilis (B. subtilis), Escherichia coli (E. coli), Candida albicans (C. albicans), Aspergillus oryzae (A. oryzae). Furthermore, the extract did not affect the inflammatory cytokine production of interleukin-10 and interferon-${\gamma}$ in NK92 cells. From the above results, we found that R. formosa extract has considerable antioxidant and elastase inhibitory effects, and does not stimulate immune cells. These findings suggest that R. formosa extract may be used as a bioactive component in cosmetic composition.

Evaluation of Sprouted Barley as a Nutritive Feed Additive for Protaetia brevitarsis and Its Antibacterial Action against Serratia marcescens (흰점박이꽃무지 사료첨가제로서 새싹보리의 곤충병원성 세균에 대한 항균 효과에 관한 연구)

  • Song, Myung Ha;Kim, Nang-Hee;Park, Kwan-Ho;Kim, Eunsun;Kim, Yongsoon
    • Journal of Life Science
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    • v.31 no.5
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    • pp.475-480
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    • 2021
  • Interest in edible insects such as Protaetia brevitarsis has increased rapidly, and several insect producers use these insects in industrialized mass production. However, mass rearing of insects can cause insect diseases. Sprouted barley is a valuable source of nutrients and has antioxidant, antimicrobial, anti-inflammatory, and anti-cancer effects. This study was conducted to investigate the effect of sprouted barley as a feed additive for producing healthy P. brevitarsis larvae. P. brevitarsis larvae were fed feeds with or without sprouted barley, and their body weight and larval period wewe checked weekly. To confirm the antibacterial effects of sprouted barley, in vitro bioassays were performed by counting Serratia marcescens colonies, and in vivo bioassays were performed by determining the survival rate and body weights of the S. marcescens-infected larvae. Larvae fed different feeds were analyzed for their nutrient compositions (i.e., such as proximate composition, minerals, amino acids, and heavy metals). Larvae fed 5% and 10% sprouted barley had maximum weight increases of 19.2% and 23.1%, respectively. Both treatment groups had significantly shorter larval periods than those of the control group. Sprouted barley markedly inhibited the growth of entomopathogenic S. marcescens. Furthermore, larvae fed sprouted barley exhibited higher Cu, Zn, and K levels. Seventeen amino acids were present in larvae fed sprouted barley, of which, tyrosine and glutamic acid were predominant. No heavy metals were detected in any of the investigated groups. Therefore, sprouted barley may be a suitable feed additive for producing high-quality P. brevitarsis larvae.

β-glucan Stimulates Release of TNF-α in Human Monocytic THP-1 Cells (인간 단핵구 THP-1 세포에서 β-glucan으로 인한 TNF-α 분비 증가 효과)

  • Keum, Bo Ram;Hyeon, Jin Yi;Choe, So Hui;Jin, Ji Young;Jeong, Ji Woo;Lim, Jong Min;Park, Dong Chan;Cho, Kwang Keun;Choi, Eun Young;Choi, In Soon
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1256-1261
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    • 2017
  • ${\beta}$-glucan is a constituent of the cell wall of fungi, yeast and plants. It plays an important role in the immune system such as activation of immunocyte, release of pro-inflammatory and anti-cancer effect. The immune system maintains a healthy immune homeostasis. However, when pathogenic substances enter the body, immune homeostasis can break down and disease can be triggered. Therefore, we studied a substance that regulates immune homeostasis. The purpose of the study we demonstrated whether the ${\beta}$-glucan can be applied to the immune-modulation effects in human monocytic THP-1 cells. ${\beta}$-glucan was incubated in THP-1 cells at various concentrations. The $TNF-{\alpha}$ mRNA expression and protein levels were analyzed by ELISA and Real-time PCR. Additionally, the expression of MAPKs (p38 and JNK), $I{\kappa}B-{\alpha}$ and $NF-{\kappa}B$ p50 were analyzed by western blot. ${\beta}$-glucan enhanced the production of $TNF-{\alpha}$ mRNA expression and protein levels in human monocytic THP-1 cells. In addition, activation of MAPKs (p38 and JNK) and $NF-{\kappa}B$ p50 induced by ${\beta}$-glucan were increased. The study suggests that ${\beta}$-glucan contributes to immune-stimulation effect by production $TNF-{\alpha}$ in human monocytic THP-1 cells, and that MAPKs and $NF-{\kappa}B$ p50 are involved in the process. Synthetically, we have suggested ${\beta}$-glucan may be improved to immune system effect in human monocytic THP-1 cells.

A Study on Bone Formation & Osteoporosis by Taeyoungion-Jahage Extracts

  • Kim, Yi-Geun;Seong, Jun-Ho;Kim, Dong-Il;Lee, Tae-Kyun;Kim, Jun-Ki;Park, Young-Duck
    • The Journal of Korean Obstetrics and Gynecology
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    • v.15 no.4
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    • pp.45-60
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    • 2002
  • Mouse calvarial osteoblast cells were isolated and cultured. To examine whether the cells produce active gelatinases in culture medium or not,the cells were analyzed using by zymograsphic analysis on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). We show that mouse calvarial osteoblasts in culture constitutively synthesize progelatinase- A. Then, mouse osteoblasts, which were stimulated by PTH, $1,25(OH)_2D_3$, mononuclear cell conditioned medium (MCM) and IL-1 as bone resorption agent's, showed increased collagenolysis by producing the active gelatinase. However, treatment of indomethacin and dexamethasone significantly decreased those effects of collagenolysis in mouse osteoblastic cells. On the other hand, IL-1 in stimulating bone resorption was examined using fetal mouse long bone organ culture. IL-1 stimulated bone resorption and produced marked resorption when present simultaneously. Furthermore, when it was examined the effects of indomethacin and dexamethasone on the dose dependent responses of $IL-1{\alpha}$, indomethacin and dexametasone produced a rightward shift in the IL-1 dose response curve. The results of in vitro cytotoxicities showed that Taeyoungjon-Jahage water extracts(T.Y.J-J.H.G extracts) have no any cytotoxicities in concentrations of $1-200\;{\mu}g/ml$ and furthermore there is no any cytotoxicity even in concentration of $300\;{\mu}g/ml$ on mouse calvarial bone cells. T.Y.J.-J.H.G. extracts had protective activity against PTH (2 units/mI), or MCM (5%, v/v), or $rhIL-1{\alpha}$ (1 ng/mI) or $1,25(OH)_2D3$ (10 ng/ml) , $IL-1{\alpha}$ and $IL-1{\beta}-induced$ collagenolysis in the mouse calvarial cells. Pretreatment of the T.Y.J.-J.H.G.extracts for 1 h, which by itself had little effect on cell survival, did not enhance the collagenolysis, nor significantly reduced the collagenolysis by pretreatment. Furthermore. the medicinal extracts were shown to have the protective effects against collagenolysis induced by $IL-1{\alpha}$ and $IL-1{\beta}$. Pretreatment of the extracts for 1 h significantly reduced the collagenolysis. Interestingly, the T.Y.J.-J.H.G. extracts were shown to have the inhibiting effects against gelatinase enzyme and processing activity induced by the bone resortion agents of PTH, $1,25(OH)_2D_3$, $IL-1{\beta}$ and $IL-1{\alpha}$, with strong protective effect in pretreatment with the extracts. T.Y.J.-J.H.G. extracts were shown to have the inhibiting effects against $IL-1{\alpha}-$ and $IL-1{\beta}-stimulated$ bone resorption and the effect of the pretreatment with a various concentrations of the medicinal extracts were significant. The inhibition extent and phenomena of IL-1 stimulated bone resorption by nonsteroidal anti-inflammatory agents of indomethacin and dexamethasone were similar to those obtained by T.Y.J.-J.H.G. extracts treatment in the mouse calvarial tissue culture system. These results indicated that the T.Y.J.-J.H.G.-water extracts are highly stable and applicable to clinical uses in osteoporosis.

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Studies of Xanthium strumarium Extract Suppressing Th17-cell Differentiation and Anti-dermatitic Effect in BMAC-induced Atopy Dermatitis of NC/Nga Mice (창이자 추출물이 아토피 피부염 유발 생쥐의 비장 세포 Th17의 세포분화 억제에 따른 아토피 피부 상태에 미치는 영향)

  • Kim, Kum-Lan;Choe, Tae-Boo
    • KSBB Journal
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    • v.24 no.4
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    • pp.383-392
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    • 2009
  • Xanthii fructus which is well known as "Chang-ihjah" in Korea is the dried fruit of Xanthium strumarium L. (or Xanthium sibiricum PATR. Ex WIDD., Asteraceae. XS). Water extract of this fruit has been used for treatment of various inflammatory diseases such as tympanitis, allergic rhinitis, or ozena as alternative therapy material usually by oral administration in far Eastern countries including Korea. In this study, the effect of XS extract (XS-E) or XS-30% acetone fraction layer (XS-30% AFL) on the differentiation of $CD4^+$ T cells isolated from NC/Nga mouse and the production of IL-17 was investigated. The experimental results showed that $100\;{\mu}g$/mL of XS-E could decrease the production of IL-17 by $CD4^+$ Th17 cells by 2 fold and only $20\;{\mu}g$/mL of XS-30% AFL could inhibit 3.5 fold. The amount of IL-17A and IL-22 mRNA determined by real-time PCR was decreased remarkably when XS-E or XS-30% AFL was treated on $CD4^+$ Th17 cells(p<0.01, p<0.001). The amount of IL-17A protein determined by ELISA was also decreased remarkably(p<0.05, p<0.001). To study the effect of XS-E or XS-30% AFL on the proliferation of Th17 cells, $CD4^+$ T cells of a NC/Nga mouse was firstly differentiated by rIL-6/TGF-$\beta$ and then stimulated by rIL-23. The control group of Th17 cells were doubled every each day, while those of XS-E or XS-30% AFL treated group were shown to be delayed remarkably by these extracts. In conclusion, XS can inhibit the differentiation of Th17 cells of NC/Nga mouse and the production of IL-17 successfully, which may be a beneficial result for the treatment of atopic skin dermatitis.

Apoptotic Effect of Pinosylvin at a High Concentration Regulated by c-Jun N-Terminal Kinase in Bovine Aortic Endothelial Cells (혈관내피세포에서 c-Jun N-terminal kinase에 의해 조절되는 세포사멸에 고농도의 피노실빈이 미치는 효과)

  • Song, Jina;Park, Jinsun;Jeong, Eunsil;So, A-Young;Pyee, Jaeho;Park, Heonyong
    • Journal of Life Science
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    • v.25 no.4
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    • pp.416-424
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    • 2015
  • Pinosylvin is a stilbenoid found in the Pinus species. Pinosylvin at ~pM to ~nM concentrations induces cell proliferation, cell migration and anti-inflammatory activity in endothelial cells. However, it was recently reported that pinosylvin at high concentrations (50 to 100 μM) induces cell death in bovine aortic endothelial cells. In this study, we conducted a series of experiments to discover how pinosylvin at a high concentration (50 μM) induces endothelial cell death. Pinosylvin at the high concentration was shown to induce endothelial cell apoptosis through enhancing caspase-3 activity, flip-flop of phosphatidyl serine, and nuclear fragmentation. We found that pinosylvin at the high concentration additively increased caspase-3 activity enhanced by serum-starvation or treatment with 100 μM etoposide. We also determined that pinosylvin at the high concentration promoted activations of c-Jun N-terminal kinase (JNK) and endothelial nitric oxide synthetase (eNOS). We further ran a series of experiments to find out which signaling molecule plays a critical role in the pinosylvin-induced apoptosis. We finally found that SP-600125, a JNK inhibitor, had an inhibitory effect on the pinosylvin-induced endothelial cell death, but L-NAME, an eNOS inhibitor, had no effect. These data indicate that JNK is involved in the pinosylvin-induced apoptosis. Collectively, pinosylvin at high doses induces cell apoptosis via JNK activation.

Inhibition of Lipopolysaccharide-Inducible Nitric Oxide Synthase, $TNF-{\alpha}$, $IL-1{\beta}$ and COX-2 Expression by Flower and Whole Plant of Lonicera japonica (금은화(金銀花) 및 금은화전초(金銀花全草)가 Raw 264.7 cell에서 LPS로 유도된 NO의 생성, iNOS, COX-2 및 cytokine에 미치는 영향)

  • Lee, Dong-Eun;Lee, Jae-Ryung;Kim, Young-Woo;Kwon, Young-Kyu;Byun, Sung-Hui;Shin, Sang-Woo;Suh, Seong-Il;Kwon, Taeg-Kyu;Byun, Joon-Seok;Kim, Sang-Chan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.2
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    • pp.481-489
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    • 2005
  • Lonicerae Flos has antibacterial effects against Staphylococcus aureus, streptococci, pneumococci, Bacillus dysenterii, Salmonella typhi, and paratyphoid. It is an antiviral agent. The herb has a cytoprotective effect against $CCl_{4}-induced$ hepatic injury. It has antilipemic action, interfering with lipid absorption from the gut. Nowadays this herb is used mainly in the treatment of upper respiratory infections, such as tonsillitis and acute laryngitis. It is also used in the treatment of skin suppurations, such as carbuncles, and to treat viral conjunctivitis, influenza, pneumonia, and mastitis. Lonicerae Flos is dried flower buds of Lonicera japonica, L. hypoglauca, L. confusa, or L. dasystyla. But, for the most part, we use whole plant of Lonicera japonica, as a flower bud of it. And, little is known of the original copy of effects of whole plant, except for the 'Bon-Cho-Gang-Mok', which is written the effects of flower of Lonicera japonica are equal to effects of leaves and branch of it. The present study was conducted to evaluate the effect of flower and whole plant of Lonicera japonica on the regulatory mechanism of cytokines, inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2) for the immunological activities in Raw 264.7 cells. In Raw 264.7 cells stimulated with lipopolysaccharide (LPS) to mimic inflammation, flower and whole plant of Lonicera japonica water extracts inhibited nitric oxide production in a dose-dependent manner and abrogated iNOS and COX-2. Flower and whole plant of Lonicera japonica water extract did not affect on cell viability. To investigate the mechanism by which flower and whole plant of Lonicera japonica water extract inhibits iNOS and COX-2 gene expression, we examined the on phosphorylation of inhibitor ${\kappa}B{\alpha}$ and assessed production of $TNF-{\alpha}$, $interleukin-1{\beta}$ $(IL-1{\beta})$ and interleukin-6 (IL-6). Results provided evidence that flower and whole plant of Lonicera japonica inhibited the production of $IL-1{\beta}$, IL-6 and activated the phosphorylation of inhibitor ${\kappa}B{\alpha}$ in Raw 264.7 cells activated with LPS. These findings suggest that flower and whole plant of Lonicera japonica can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections, respectively.

The Study of anti-inflammatory Mechanism with Cobra Venom on Astrocytes of Rats (뇌(腦) 성상세포(星狀細胞)를 대상으로 한 Cobrotoxin의 염증(炎症) 치료(治療) 기전(機轉) 연구(硏究))

  • Yoo, Jae-ryong;Song, Ho-sueb
    • Journal of Acupuncture Research
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    • v.22 no.3
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    • pp.155-167
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    • 2005
  • Objectives : The purpose of this study was to investigate the anti-inflammatory effect of Cobrotoxin on binding affinity of cobrotoxin with P50, $IKK{\alpa}$ and $IKK{\beta}$, activities of NF-${\kappa}B$, Cell viability of astrocyte, expressions of protein molecules of NF-${\kappa}B$ such as P50, P-$1{kappa}B$, $1{\kappa}B$ and iflammation related genes such as Cox-2, iNOS, cPLA2 in the SNP or LPS induced Inflammatory pathway of Rats' astrocytes. Methods : In this study, The expression of cytosolic phospholipase A2, Nitric oxcide, Cyclooxygenase-2 and inducible nitrogen oxide synthase was determined by western blotting with corresponding antibodies, and the generation of NF-${\kappa}B$ was assayed by EMSA method in astrocytes of rats. The Cell viability of astrocytes was determined by MTT assay, and Binding affinity of Cobrotoxin with P50, $IKK{\alpha}$ and $IKK{\beta}$ was assayed by Surface plasmon resonance analysis, and NF-${\kappa}B$ dependent luciferase activity was determined by luciferase analysis, and Uptake of cobrotoxin in astrocytes was identified by Confocal laser scanning microscope Results : 1. Compared with control, LPS-induced NF-${\kappa}B$ DNA binding activity was decreased significantly by 0.1, $0.5{\mu}g/m{\ell}$ of Cobrotoxin in Astrocyte. 2. Compared with control, LPS-induced NF-kB dependent luciferase expression was decreased significantly by 0.1, 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin in Astrocyte. 3. Compared with control, SNP induced P50, $I{\kappa}B$ expressions in astrocyte were decreased significantly by 0.1, 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin and P-$1{\kappa}B$ expression was decreased significantly by 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin. 4. Compared with control, LPS induced P50, $1{\kappa}B$ expressions in astrocyte were decreased significantly by 0.5 and $1{\mu}g/m{\ell}$ of Cobrotoxin. 5. Compared with control, SNP induced Cox-2, iNOS, CPLA2 expressions in astrocyte were decreased significantly by $1{\mu}g/m{\ell}$ of Cobrotoxin. 6. Compared with control, LPS induced Cox-2, cPLA2 expressions in astrocyte were decreased significantly by 0.1, 0.5, $1{\mu}g/m{\ell}$ of Cobrotoxin and iNOS expression was decreased significantly by 0.5, $1{\mu}g/m{\ell}$ of Cobrotoxin. 7. Compared with $0.5{\mu}g/m{\ell}$ of Cobrotoxin, SNP-induced NF-${\kappa}B$ DNA bindins activity in astrocyte was increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM. 8. Compared with $0.5{\mu}g/m{\ell}$ of Cobrotoxin, LPS-induced NF-${\kappa}B$ DNA binding activity in astrocyte was increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM, Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM, Cobrotoxin $0.5{\mu}g/m{\ell}$with GSH 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 5mM 9. Compared with $0.1{\mu}g/m{\ell}$ of cobrotoxin, SNP induced P50 expressions in astrocyte were increased significantly by Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 1mM, Cobrotoxin $0.5{\mu}g/m{\ell}$ with DTT 5mM Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 1mM and Cobrotoxin $0.5{\mu}g/m{\ell}$ with GSH 5mM. 10. The uptake of the labeled cobrotoxin into the cells was shown under a confocal laser scanning microscope. cobrotoxin was uptaken into the membrane and nucleus of astrocytes. Conclusions : In summary, the present results demonstrate that cobrotoxin directly binds to sulfhydryl group of p50 and IKKS resulting In the reduction of translocation of p50 and IkB release, thereby inhibits activation of NF-${\kappa}B$, and suggest that pico to nanomolar range of cobrotoxin could inhibit the expression of genes in the NF-${\kappa}B$ signal pathway.

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