• Title/Summary/Keyword: Nitric oxide reduction

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Evaluation of Biological Activity of Veronica incana Extracts (Veronica incana 추출물의 생물학적 활성 평가)

  • Mi-Rae Shin;Mi Yeong Yoon;Min Ju Kim;Il-Ha Jeong;Hui Yeon An;Ji-Won Jung;Seong-Soo Roh
    • The Korea Journal of Herbology
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    • v.39 no.3
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    • pp.57-67
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    • 2024
  • Objectives : The aim of this study is to evaluate the potential biological activity of Veronica incana extracts (VIE) through in vitro, ex vivo, and in vivo experiments. Methods : In vitro, we conducted analyses on the total polyphenol (TP) and total flavonoid (TF) levels, alongside DPPHand ABTS radical scavenging activities. Ex vivo evaluations on adipose tissue measured glycerol release as a marker of lipolysis. In LPS-induced RAW 264.7 cells, we quantified nitric oxide (NO) production. Following H2O2 induction in U2OS cells, we performed mitochondrial assays such as MitoSox and MitoTracker. Moreover, Bodipy assays were conducted in 3T3-L1 cells. In vivo, we performed anti-osteoarthritis effect of VIE against monosodium iodoacetate (MIA)-induced osteoarthritis in rats. Results : The results presented encompass a myriad of models, from cell culture to animal experiments as well as ex vivo studies. VIE demonstrated high TP and TF contents, potent DPPH and ABTS scavenging activities, and regulated glycerol release. Moreover, the inhibition of NO production in LPS-induced inflammation was notably confirmed and the reduction of lipid droplets was distinctly shown. Furthermore, in H2O2-induced U2OS cells, MitoSox was effectively reduced while MitoTracker noticeably increased. In vivo assays confirmed a significant increase in hindpaw weight distribution (HWD) decreased by MIA after VIE treatment. Additionally, VIE inhibited serum inflammatory cytokines (TNF-𝛼, IL-6, and IL-1𝛽) and MDA levels in joint tissue. Conclusion : In conclusion, Veronica incana exhibited various pharmacological effects including antioxidant, anti-obesity, and anti-inflammatory properties.

Protective effects of Gastrodia rhizoma and steamed & fermented Gastrodiae rhizoma with anti-oxidant efficacy and suppression of NFκB signaling pathway on LPS-induced liver injury (LPS로 유발한 간손상 마우스에서 항산화 및 항염증 효능을 통한 천마와 증숙 발효 천마의 간보호 효과)

  • Kim, Hyoung-Jin;Kwon, O Jun;Lee, Ah Reum;Roh, Seong-Soo;Seo, Young-Bae
    • Journal of Applied Biological Chemistry
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    • v.59 no.3
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    • pp.179-188
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    • 2016
  • This study is aimed to evaluate the protective effect of Gastrodiae rhizoma and steamed, dried & fermented Gastrodiae rhizoma on Lipopolysaccharide (LPS)-induced hepatic injury in the mice model. Sample was selected to GR0F0 (not processed gastrodia rhizome) and GR6F4 (fermented with Saccharomyces cerevisiae before steamed and dried 6 times) based on 1,1-diphenyl-2-picrylhydrazyl, 2,2'-azinobis-3-ethyl-benzothiazoline-6-sulfonic acid, and High-performance liquid chromatography analysis. Mice were randomly divided into 4 groups - Normal group, vehicle group (LPS treated), GR0F0 group (fed GR0F0 before LPS treated) and GR6F4 group (fed GR6F4 before LPS treated) with 6 mice in each group. GR0F0 group and GR6F4 group were fed each extract 200 mg/kg/day during 8 days. LPS 20 mg/kg injected to the experimental groups as abdominal injection. We measured aspartate aminotransferase, alanine amino-transferase in serum. GR0F0 and GR6F4 showed a significant decrease compared to the vehicle group. As a result of measuring the ROS, GR6F4 group showed a significant reduction in both the serum and liver tissues compared to the vehicle group. GR0F0 group showed a significant reduction only in the liver tissues. Activator protein-1, cyclooxygenase-2, and Inducible nitric oxide synthase were significantly decreased GR0F0 group and GR6F4 group. But tumor necrosis factor alpha only showed a significant reduction in GR6F4 group. GR0F0 and GR6F4 groups against liver damage in mice with LPS. That showed significant effects on anti-oxidant and anti-inflammatory action. The effects of GR6F4 group showed superior results compared to GR0F0 group. Therefore, Steamed, dried & fermented Gastrodia rhizoma was might have a protective effect on liver injury.

In vitro Antioxidant and Anti-inflammatory Effects of Ethanol Extracts from Sprout of Evening Primrose (Oenothera laciniata) and Gooseberry (Actinidia arguta) (달맞이순과 다래순 에탄올 추출물의 in vitro 항산화효과 및 항염증효과)

  • Kwak, Chung Shil;Lee, Ji Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.2
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    • pp.207-215
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    • 2014
  • To investigate the biological benefits of Korean traditional vegetables, anti-oxidative and anti-inflammatory effects of ethanol extracts from blanched and dried sprouts of evening primrose (Oenothera laciniata, OL) and gooseberry (Actinidia arguta, AA) were measured. Total polyphenol and flavonoid contents of OL were higher than those of AA; OL contained 60.4 mg tannic acid/g dry weight and 31.9 mg rutin/g dry weight, while AA contained 33.0 mg tannic acid/g dry weight and 20.3 mg rutin/g dry weight. The $IC_{50}$ value for DPPH radical scavenging activity was $58.2{\mu}g/mL$ for OL ethanol extract and $122.1{\mu}g/mL$ for AA ethanol extract. The reducing power upon $500{\mu}g/mL$ of ethanol extract treatment was as strong as $52.1{\mu}g$ ascorbate eq./mL for OL and $45.3{\mu}g$ ascorbate eq./mL for AA. Regarding anti-inflammatory effects, inhibition rate against 5-lipoxygenase (LOX) and cyclooxygenase (COX)-2 activities were 29.5% and 79.5% for OL, as well as 11.5% and 39.1% for AA, respectively at a concentration of $250{\mu}g/mL$. Lipopolysaccaride ($1{\mu}g/mL$)-treated RAW 264.7 macrophage cells subjected to OL ethanol extract at various concentrations ($0{\sim}25{\mu}g/mL$) showed significantly reduced synthesis of nitrite oxide (NO), prostaglandin (PG) E2, and IL-6 in a dose-dependent manner without cytotoxicity, although TNF-${\alpha}$ synthesis was not affected. In conclusion, both OL and AA sprouts showed strong antioxidative activity, whereas OL showed very strong anti-inflammatory activity via effective reduction of NO, PGE2, and IL-6 synthesis in LPS-activated macrophage cells.

Kinetics of Nitric Oxide Reduction with Alkali Metal and Alkali Earth Metal Impregnated Bamboo Activated Carbon (알칼리금속과 알칼리 토금속 촉매 담지 대나무 활성탄의 NO 가스 반응 특성)

  • Bak, Young-Cheol;Choi, Joo-Hong
    • Korean Chemical Engineering Research
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    • v.54 no.5
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    • pp.671-677
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    • 2016
  • The impregnated alkali metal (Na, K), and the alkali earth metal (Ca, Mg) activated carbons were produced from the bamboo activated carbon by soaking method of alkali metals and alkali earth metals solution. The carbonization and activation of raw material was conducted at $900^{\circ}C$. The specific surface area and the pore size distribution of the prepared activated carbons were measured. Also, NO and activated carbon reaction were conducted in a thermogravimetric analyzer in order to use for de-NOx agents of the used activated carbon. Carbon-NO reactions were carried out in the nonisothermal condition (the reaction temperature $20{\sim}850^{\circ}C$, NO 1 kPa) and the isothermal condition (the reaction temperature 600, 650, 700, 750, 800, $850^{\circ}C$, NO 0.1~1.8 kPa). As results, the specific volume and the surface area of the impregnated alkali bamboo activated carbons were decreased with increasing amounts of the alkali. In the NO reaction, the reaction rate of the impregnated alkali bamboo activated carbons was promoted to compare with that of the bamboo activated carbon [BA] in the order of BA(Ca)> BA(Na)> BA(K)> BA(Mg) > BA. Measured the reaction orders of NO concentration and the activation energy were 0.76[BA], 0.63[BA(Na)], 0.77[BA(K)], 0.42[BA(Ca)], 0.30 [BA(Mg)], and 82.87 kJ/mol[BA], 37.85 kJ/mol[BA(Na)], 69.98 kJ/mol[BA(K)], 33.43 kJ/mol[BA(Ca)], 88.90 kJ/mol [BA(Mg)], respectively.

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.

L-AHG-mediated Suppression of M1 Polarization and Pro-inflammatory Signaling Pathways in LPS-stimulated RAW264.7 Macrophages (LPS에 의해 자극된 RAW264.7 대식세포에서 L-AHG에 의한 M1 분극화 및 친염증 신호 경로의 억제)

  • Won Young Jang;Shin Young Park;Ki Youn Kim;Do Youn Jun;Young-Seuk Bae;Young Ho Kim
    • Journal of Life Science
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    • v.34 no.7
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    • pp.443-452
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    • 2024
  • This study aimed to examine the influence of 3,6-anhydroxygalactose (L-AHG) on the pro-inflammatory M1 polarization and pro-inflammatory responses observed in the RAW264.7 mouse macrophage cell line following stimulation with lipopolysaccharides (LPS). L-AHG exhibited a significant and dose-dependent inhibition of inducible nitric oxide synthase (iNOS) expression, a hallmark of M1 polarization, and subsequent NO production in LPS-stimulated RAW264.7 cells. Furthermore, the LPS-induced upregulation of cyclooxygenase-2 (COX-2), which drives the production of prostaglandin E2, an inflammatory mediator, was also inhibited by L-AHG. L-AHG did not affect the LPS-triggered Toll-like receptor 4 (TLR4)-mediated pro-inflammatory signaling pathway, which culminated in the activation of transforming growth factor-β-activated kinase 1 (TAK1). However, it was observed to inhibit the generation of reactive oxugen species (ROS) in a dose-dependent manner, as well as the TAK1-driven activation of JNK and p38 MAPK. Given that the active p38 MAPK is known to contribute to the assembly of active nicotinamide adenine dinucleotide phosphate (NADPH) oxidase, which catalyzes the intracellular generation of pro-inflammatory ROS in LPS-stimulated macrophages, the dose-dependent reduction in the LPS-induced ROS generation by L-AHG may be mainly due to the prevention of TAK1-driven activation of p38 MAPK. Together, these results demonstrate that the L-AHG-mediated inhibition of the TAK1-JNK/p38 MAPK activation phase of the pro-inflammatory signaling pathway in LPS-stimulated RAW264.7 cells by L-AHG represents a promising mechanism for suppressing M1 polarization and pro-inflammatory responses in macrophages.

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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