• Title/Summary/Keyword: NO/iNOS activity

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Evaluation of the Cosmeceutical Activity of Apocynum lancifolium Russanov Extracts (개정향풀 추출물의 향장학적 활성 평가)

  • Park, Ji Hyo;Lee, Ji-An
    • Journal of Convergence for Information Technology
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    • v.11 no.1
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    • pp.236-243
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    • 2021
  • In this study, the functional cosmetic activity of hot-water (AW) and methanol(AM) extracts from Apocynum lancifolium Russanov are examined. In the DPPH antioxidant activity test, the AW extract showed the highest inhibition rate of 90.5% (IC50 37.717 ± 8.209 ㎍/mL) and the antioxidative activity of ABTS showed a high activity in the AW extract with an IC50 of 185.244 ± 12.602 ㎍/mL and 96.2%, respectively. In RAW264.7 macrophages, the two extracts significantly suppressed the LPS-induced nitric oxide (NO), TNF-α, IL-6 production and iNOS expression level. The MTT assay measured by the cell survival rate showed that AW and AM extracts have no toxicity. The astringent activity of the AW extract exhibited the highest astringent activity with 74.336±2.487 mg/mL. Therefore, these results suggest that A. lancifolium extracts have a health promotion potential which can be further developed as food additive, natural products for cosmetic purpose.

Anti-inflammatory and Anti-bacterial Constituents from the Extracts of Daucus carota var. sativa Aerial Parts (당근 지상부 추출물 유래 항염 및 항균 활성 성분)

  • Kim, Jung Eun;Jo, Yeon Jeong;Lee, Nam Ho
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.44 no.4
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    • pp.427-436
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    • 2018
  • In this study, we investigated anti-inflammatory and anti-bacterial constituents from Daucus carota var. sativa (carrot) areal parts. For the extract and solvent fractions, the anti-inflammatory activities were examined by measuring the nitric oxide (NO) production using LPS-stimulated RAW 264.7 cells. Among them, the ethyl acetate (EtOAc) fraction decreased the NO level in a dose-dependent manner. To elucidate further anti-inflammatory mechanisms, EtOAc fraction was evaluated by estimating their effects on the production of prostaglandin $E_2$ and pro-inflammatory cytokines as well as on the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). As a result, the EtOAc fraction was determined to inhibit the production of $PGE_2$, IL-$1{\beta}$, IL-6 and reduce the iNOS, COX-2 protein expression. Upon the anti-bacterial tests using Staphylococcus epidermidis and Propionibacterium acnes, n-hexane (Hex) and EtOAc fractions showed the most potent activities. Three phytochemicals were isolated form the EtOAc fraction; diosmetin (1), diosmin (2), cynaroside (3). The chemical structures of the isolated compounds were elucidated based on the spectroscopic data including $^1H$ and $^{13}C$ NMR spectra, as well as comparison of the data to the literatures. Anti-inflammatory and anti-bacterial effects were studied for the isolates. All of the compounds (1 - 3) decreased the NO production, effectively. Also, compound 3 showed anti-bacterial activity on P. acnes. Based on these results, D. carota var. sativa extract could be potentially applicable as anti-inflammatory and anti-bacterial ingredients in cosmetic formulations.

Effect of Trigonel/ae Semen on Peroxynitrite Scavenging Activity and Protein Expression of Proinflammatory Mediators (호로파(葫蘆巴)추출물의 Peroxynitrite 제거 및 염증 인자 단백질 억제 효과)

  • Lee, Cheol-Ho;Jeong, Ji-Cheon
    • The Journal of Internal Korean Medicine
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    • v.27 no.3
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    • pp.688-697
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    • 2006
  • Objectives : Peroxynitrite ($ONOO^-$), $O_2^-$ and nitric oxide (NO) are cytotoxic species that can oxidize several cellular components such as proteins. lipids and DNA. It has been implicated in the aging process and age-related disease such as Alzheimer's disease, rheumatoid arthritis, cancer and atherosclerosis. The aim of this study was to investigate $ONOO^-$ scavenging activities. and that of its precursors. NO and $O_2^-$ of Trigonellae Semen. Methods : To investigate $ONOO^-$. NO. $O_2^-$ scavenging activities, fluorescent probes, namely 2'.7'-dichlorodihydrofluorescein diacetate (DCFDA). 4.5-diaminofluorescein (DAF-2) and dihydrorhodamine 123 (DHR 123) were used. Protein expression levels of iNOS, COX-2, NF-${\kappa}B$, and VCAM-1 were assayed by western blot. Results : Trigonellae Semen markedly scavenged authentic $ONOO^-$, $O_2^-$ and NO. It also inhibited $ONOO^-$ induced by $O_2^-$ and NO which are derived from SIN-1. Furthermore. Trigonellae Semen inhibited $ONOO^-$, $O_2^-$ and NO generation in LPS-treated ICR mouse kidney postmitochondria. Trigonellae Semen inhibited gene expression of iNOS, COX-2, VCAM-l and NF-${\kappa}B$ (p65) activation. Conclusions : These results suggest that Trigonellae Semen is an effective $ONOO^-$, $O_2^-$ and NO scavenger. and that this substance has a potential role as an inhibitor of the aging process, and in therapy against age-related diseases.

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TI-I-174, a Synthetic Chalcone Derivative, Suppresses Nitric Oxide Production in Murine Macrophages via Heme Oxygenase-1 Induction and Inhibition of AP-1

  • Kim, Mi Jin;Kadayat, Taraman;Kim, Da Eun;Lee, Eung-Seok;Park, Pil-Hoon
    • Biomolecules & Therapeutics
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    • v.22 no.5
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    • pp.390-399
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    • 2014
  • Chalcones (1,3-diaryl-2-propen-1-ones), a flavonoid subfamily, are widely known for their anti-inflammatory properties. Propenone moiety in chalcones is known to play an important role in generating biological responses by chalcones. In the present study, we synthesized chalcone derivatives structurally modified in propenone moiety and examined inhibitory effect on nitric oxide (NO) production and its potential mechanisms. Among the chalcone derivatives used for this study, TI-I-174 (3-(2-Hydroxyphenyl)-1-(thiophen-3-yl)prop-2-en-1-one) most potently inhibited lipopolysaccharide (LPS)-stimulated nitrite production in RAW 264.7 macrophages. TI-I-174 treatment also markedly inhibited inducible nitric oxide synthase (iNOS) expression. However, TI-I-174 did not significantly affect production of IL-6, cyclooxygenase-2 (COX-2) and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), implying that TI-I-174 inhibits production of inflammatory mediators in a selective manner. Treatment of macrophages with TI-I-174 significantly inhibited transcriptional activity of activator protein-1 (AP-1) as determined by luciferase reporter gene assay, whereas nuclear factor-${\kappa}B$ (NF-${\kappa}B$) activity was not affected by TI-I-1744. In addition, TI-I-174 significantly inhibited activation of c-Jun-N-Terminal kinase (JNK) without affecting ERK1/2 and p38MAPK, indicating that down-regulation of iNOS gene expression by TI-I-174 is mainly attributed by blockade of JNK/AP-1 activation. We also demonstrated that TI-I-174 treatment led to an increase in heme oxygenase-1 (HO-1) expression both at mRNA and protein level. Transfection of siRNA targeting HO-1 reversed TI-I-174-mediated inhibition of nitrite production. Taken together, these results indicate that TI-I-174 suppresses NO production in LPS-stimulated RAW 264.7 macrophages via induction of HO-1 and blockade of AP-1 activation.

Anti-inflammatory Effects of Lemon Myrtle (Backhousia citriodora) Leaf Extracts in LPS-induced RAW 264.7 Cells (LPS로 유도된 RAW 264.7 세포에 대한 레몬 머틀 잎 추출물의 항염증 효과)

  • Kim, Pan Kil;Jung, Kyung Im;Choi, Young Ju;Gal, Sang Wan
    • Journal of Life Science
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    • v.27 no.9
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    • pp.986-993
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    • 2017
  • Lemon myrtle (Backhousia citriodora) has been identified as one of the plants that are likely to undergo important commercial exploitation. This study was carried out to investigate the anti-inflammatory activities and nitric oxide synthase (iNOS) expression of hot water (LMW) and 80% ethanol (LME) extracts from lemon myrtle leaf in lipopolysaccharide-induced (LPS) RAW 264.7 cells. The total phenol content of LMW and LME was 207.44 and $331.54{\mu}g$ tannic acid equivalents (TAE)/mg, respectively (p<0.01). DPPH radical scavenging activities of LMW and LME were remarkably increased in a dose-dependent manner, and were about 90.69% and 92.50% at 0.5 mg/ml, respectively. Superoxide dismutase (SOD) activities of LMW and LME were 106.22% and 103.58% at 1 mg/ml, respectively. The highest activity (91.03%) of nitrite-scavenging was observed for LME at 1 mg/ml at pH 1.2, while the activity for LMW was about 81.03% under the same conditions (p<0.05). Anti-inflammatory effect was examined in LPS stimulated RAW 264.7 cells. Nitric oxide (NO) production were reduced to 35.41% and 78.39% by addition of LMW and LME at 0.5 mg/ml, respectively (p<0.05). LMW and LME reduced protein expression of inducible nitric oxide synthase (iNOS) in a dose-dependent manner (p<0.05). These results, we conclude that lemon myrtle may be a highly valuable natural product owing to its high-quality functional components as well as its anti-oxidant and anti-inflammatory activities.

Scrophularia Buergeriana inhibits the Production of NO through the Suppression of NF-kB adivity in LPS-stimulated Mouse Peritoneal Macrophages

  • Ha Mi Suk;Kim Young Hee;Ko Woo Shin;Kim Han Do
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.6
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    • pp.1284-1290
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    • 2002
  • Scrophularia buergeriana Miquel (Scrophulariaceae) has been used as an anti-inflammatory drug in the folk medicine recipe and been proved its anti-inflammatory effect in the oriental medicine. Since nitric oxide (NO) and superoxide anion (O/sub 2//sup -/) are ones of the major inflammatory parameters, we studied the effect of aqueous extracts of Scrophularia buergeriana (SB) on NO and O/sub 2//sup -/ production in lipopolysaccharide (LPS)-stimulated mouse peritoneal macrophages. NO, O/sub 2//sup -/production and inducible NO synthase (iNOS) level were significantly reduced in LPS-activated macrophages by SB compared to those without. Electrophoretic mobility shift assay (EMSA) indicated that SB blocked the activation of NF-kB, which was considered to be a potential transcription factor for the iNOS expression. SB also blocked degradation of IkBα. Furthermore, IkB kinase alpha (IKKα), which phosphorylates serine residues of IkB directly, is inhibited by SB. These results suggest that SB could exert its anti-inflammatory actions by suppressing the activation of NF-kB through inhibition of IKK activity.

Anti-inflammatory Effect of Inonotus obliquus Extracts in Lipopolysaccharide-induced Mouse Peritoneal Macrophage (LPS로 유도된 마우스 복강 대식세포에서 차가버섯 열수 추출물의 염증 억제 효과)

  • Ko, Suk-Kyung;Pyo, Myoung-Yun
    • Korean Journal of Pharmacognosy
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    • v.42 no.3
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    • pp.253-259
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    • 2011
  • Macrophages play a vital role in the innate immune system involving defensive cytokines such as TNF (tumor necrosis factor)-${\alpha}$ and nitric oxide (NO). Therefore, we try to elucidate the anti-inflammatory activity of Chaga mushroom (Inonotus Obliquus, IO) in murine macrophages. Raw 264.7 cells and peritoneal macrophages of mice were cultured with or without LPS/LPS + IFN-${\gamma}$ in the presence of IO aqueous extracts (IOE 0.2, 2, 20, 100 ${\mu}g$/mL) for 24 hr and 48 hr, respectively. Exposure of IOE caused the decrease of NO production and increase of TNF-${\alpha}$ production in dose-dependent manner in activated peritoneal macrophage in vitro. To further investigate anti-inflammatory effects of IO ex vivo, we orally administrated capsaicin (PC, 3 mg/kg/day) and IOE (100, 200, 400 mg/kg/day) for 4 consecutive days to C57BL/6 mice (7~9 weeks old, female), then observed the NO secretion and cytokine (TNF-${\alpha}$) production of LPS/LPS + INF-${\gamma}$-stimulated peritoneal macrophages. IOE inhibits NO secretion in dose-dependent manner both ex vivo and in vitro and increases the production of TNF-${\alpha}$ in vitro. In addition, we found that IOE possessed suppressive effects of LPS-stimulated TNF-${\alpha}$, IL-$1{\beta}$, COX-2, as well as iNOS expressions in Raw 264.7 cells. These findings indicate that IOE suppress not only the LPS-induced NO overproduction of murine peritoneal macrophages, but also iNOS, COX-2, TNF-${\alpha}$, and IL-$1{\beta}$ overexpression of LPS-induced Raw 264.7 cells. Consequently, our results suggest that IO may have the anti-inflammatory effects via suppression of the inflammatory cytokines and mediators, and be useful for the treatment of inflammatory diseases.

Anti-inflammatory Function of the Sophora japonica Extract Rutin: The Inhibitory Effect of Rutin of Korean Sophora japonica on the Productions of NO and TNF-alpha from Mouse Peritoneal Macrophages (괴화 추출물 루틴의 소염작용: 쥐의 복강대식세포로부터 NO와 TNF-alpha 생산에 있어서 괴화 루틴에 의한 억제효과)

  • Lee, Mu-Hong;Jeong, Jae-Hyun;Jeong, Myeong-Soo;Chang, Sung-Ho;Her, Erk
    • Korean Journal of Medicinal Crop Science
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    • v.18 no.2
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    • pp.105-112
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    • 2010
  • Korean Sophora japonica has been found to posses an anti-inflammatory activity. In this study, Korean Sophora japonica extract, rutin, was used to know whether rutin inhibits to produce inflammatory mediators NO and TNF-$\alpha$ from the mouse peritoneal macrophages that were treated an inflammatory agent LPS. The rutin-1 hr pretreated macrophages were incubated with LPS for 0.5~5 hrs, and then collected the supernatant and the cell lysate for measurements of the level of iNOS, NO, TNF-$\alpha$ mRNA, TNF-$\alpha$, and p-NF-${\kappa}B$. Minimal and maximal effective doses of the rutin on them were 1 and $100{\mu}g/ml$, respectively. The maximal effective dose of rutin certainly inhibted the productions of iNOS, NO, TNF-$\alpha$ mRNA, TNF-$\alpha$and p-NF-${\kappa}B$ from the LPS-treated macrophages (p<0.0001). Its $ED_{50}$ for inhibition of TNF-$\alpha$ mRNA and p-NF-${\kappa}B$ was $5{\mu}g/m{\ell}$, and for iNOS, NO, and TNF-$\alpha$ was $10{\mu}g/m{\ell}$. The rutin did not have any cytotoxic effect. As the results, the Sophora japonica rutin could be a good candidate for an anti-inflammatory action.

Effect of Trolox on Altered Vasoregulatory Gene Expression in Hepatic Ischemia/Reperfusion

  • Eum, Hyun-Ae;Lee, Sun-Mee
    • Archives of Pharmacal Research
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    • v.27 no.2
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    • pp.225-231
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    • 2004
  • This study was designed to investigate the effect of Trolox, a hydrophilic analogue of vitamin E, on the alteration of vasoregulatory gene expression during hepatic ischemia and reperfusion (I/R). Rats were subjected to 60 min of hepatic ischemia in vivo. The rats were treated intravenously with Trolox (2.5 mg/kg) or the vehicle as a control 5 min before reperfusion. Liver samples were obtained 5 h after reperfusion for a RT-PCR analysis on the mRNA for the genes of interest. These mRNA peptides are endothelin-1 (ET -1), potent vasoconstrictor peptide, its receptor $ET_A$ and $ET_B$, vasodilator endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase (iNOS), heme oxygenase-1 (HO-1), tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and cyclooxygenase-2 (COX-2). It was seen that serum alanine aminotransferase and lipid peroxi-dation levels were markedly increased after I/R and Trolox significantly suppressed this increase. In contrast, the glutathione concentration decreased in the I/R group, and this decrease was inhibited by Trolox. ET-1 mRNA expression was increased by I/R, an increase which was prevented by Trolox. The mRNA levels for $ET_A$ receptor was significantly decreased, whereas ET$_{B}$ receptor transcript increased in the I/R group. The increase in $ET_A$ was prevented by Trolox. The mRNA levels for iNOS and HO-1 significantly increased in the I/R group and Trolox attenuated this increase. There were no significant differences in eNOS mRNA expression among any of the experimental groups. The mRNA levels for COX-2 and TNF-$\alpha$ significantly increased in I/R group and Trolox also attenuated this increase. Our findings suggest that I/R induces an imbalanced hepatic vasoregulatory gene expression and Trolox ameliorates this change through its free radical scavenging activity.y.

The Experimental Study on Antioxidant, Anti-inflammatory Effects of the Boeum-jeon (BEJ) (보음전의 항산화, 항염증 효능에 관한 실험연구)

  • Su-Kyung Kim;Seong-Hee Cho;Seung-Jeong Yang
    • The Journal of Korean Obstetrics and Gynecology
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    • v.36 no.3
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    • pp.1-24
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    • 2023
  • Objectives: Boeum-jeon (BEJ) is a herbal formula composed 8 Korean medicinal herbs and is traditionally used to treat inflammatory diseases. In this study, the authors tried to confirm the antioxidant efficacy of BEJ and its anti-inflammatory effect in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Methods: In this experiment, results of BEJ on the following two were measured as follows: (1) Antioxidant effects was measured by DPPH Radical scavenging Activity, ABTS Radical scavenging Activity. (2) Anti-Inflammatory effects were evaluated by the production amount of ROS, NO, Cytokine (PGE2, IL-1β, IL-6, TNF-α), COX-2, iNOS, TNF-α, IL-1β, IL-6, HO-1, NQO1 (the previous seven are "mRNA Expression"), COX-2, iNOS, TNF-α, IL-1β, IL-6, HO-1, NQO1, NRF2 (the previous eight are "Protein Expression") ERK, JNK, p38 (the previous three are "Protein Phosphorylation") in LPS-stimulated RAW 264.7 cells. Results: The experimental measurement results are as follows: (1) DPPH Radical scavenging Activity, ABTS Radical scavenging Activity increased in a concentration -dependent manner in the BEJ-treated group. (2) As a result of measuring anti-inflammatory efficacy, the production of ROS, NO, and Cytokine (PGE2, TNF-α, IL-1β, IL-6) in the BEJ-administered group was significantly reduced compared to the control group. (3) Among mRNA Expression levels, COX-2, iNOS, IL-1β, IL-6 and TNF-α was significantly decreased in a concentration-dependent manner than in the control group, and HO-1 and NQO1 were significantly increased in a concentration -dependent manner than in the control group. (4) Among the Protein Expression levels, COX-2, iNOS, IL-1β, IL-6 and TNF-α was significantly decreased in a concentration -dependent manner compared to the control group, and HO-1, NQO1 and NRF2 was significantly increased in a concentration-dependent manner compared to the control group. (5) As a result of Protein Phosphorylation, ERK, p38 and JNK was significantly decreased compared to the control group in a concentration-dependent manner. Conclusions: Boeum-jeon has been experimentally confirmed to have antioxidant and anti-inflammatory effects, and if the evidence for efficacy is reinforced through further studies such as in vivo studies and clinical trials in the future, it can be effectively used to treat various inflammatory diseases such as bladder inflammation and chronic pelvic inflammation.