• Title/Summary/Keyword: iNOS inhibitor

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Inhibitory Effect of Sunbanghwalmyungeum MeOH Extract on Pro-inflammatory Mediator in Lipopolysaccharide - activated Raw 264.7 Cells (선방활명음(仙方活命飮)메탄올 추출물이 LPS로 유도된 Raw 264.7 Cell에서의 Pro-inflammatory Mediator에 미치는 영향)

  • Choi, Song-I;Jo, Mi-Jeong;Kim, Sang-Chan;Byun, Sung-Hui
    • The Korea Journal of Herbology
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    • v.23 no.3
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    • pp.11-18
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    • 2008
  • SunBangHwalMyungEum (SBH) has the effects of subduing swelling, resolving masses and alleviating pain in traditional oriental medicine. Recent studies showed that SunBangHwalMyungEum produced anti-cancer, anti-metastasis and immuno-modulatory effects. However there is lack of studies regarding the effects of SBH on the immunological activities. The present study was conducted to evaluate the effect of SBH on the regulatory mechanism of cytokines and nitric oxide (NO) in Raw 264.7 cells. Methods : After the treatment of SBH, cell viability was measured by MTT assay, NO production was monitored by measuring the nitrite content in culture medium. Inducible nitric oxide synthase (iNOS) was determined by immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. Results : Results provided evidence that SBH inhibited the production of NO, iNOS, $interleukin-1{\beta}$ ($(IL-1{\beta})$), IL-6, and the activation of phospholylation of inhibitor ${\kappa}B{\alpha}$ in Raw 264.7 cells activated with lipopolysaccharide. Conclusions : These findings suggest that SBH can produce anti-inflammatory effect, which may play a role in adjunctive therapy in Gram-negative bacterial infections.

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An Experimental Study on the Anti-inflammatory, Anti-pruritic and Anti-microbial Effects of the Three Herbal Prescription: Eunhwasagan-tang (EST), Sobokchukeo-tang (SCT), Wandae-tang (WDT) (대하(帶下) 처방 3종의 항염, 항소양, 항균 효능에 관한 실험 연구 : 은화사간탕(銀花瀉肝湯), 소복축어탕(少腹逐瘀湯), 완대탕(完帶湯))

  • Lee, Eun-Kyu;Park, Chan-Wook;Kim, Soo-Hyeon;Choi, Yoo-Jin;Park, Kyung-Mi;Yang, Seung-Jeong;Cho, Seong-Hee
    • The Journal of Korean Obstetrics and Gynecology
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    • v.32 no.3
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    • pp.32-56
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    • 2019
  • Objectives: The purpose of this study was to investigate the in vitro anti-inflammatory, anti-pruritic and antimicrobial effects of the three herbal prescription (EST, SCT, WDT), which has been traditionally used for treating leukorrhea induced by various infections in the female genital tract. Methods: In this experiment, the anti-inflammatory effects were evaluated by Nitric oxide (NO), $Interlukine-1{\beta}$ ($IL-1{\beta}$), Interlukine-2 (IL-2), Interlukine-6 (IL-6), Tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), Prostaglandin $E_2$ ($PGE_2$), Leukotriene $B_4$ ($LTB_4$) production amount and Inducible nitric oxide synthase (iNOS), Nuclear factor kappa B ($NF-{\kappa}B$), Cyclooxygenase-2 (COX-2) gene expression levels in RAW264.7 cells. And the anti-pruritic effects were evaluated by Histamine, Acetylcholine (ACh), Acetylcholinesterase (AChE), Substance P production amount in Mast cell/9 (MC/9) and Pheochromocytoma 12 (PC12) cells. The anti-microbial effect was measured by inhibition zone diameter on Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans and Aspergillus niger. Results: As a result of measuring anti-inflammatory efficacy, $IL-1{\beta}$, IL-2, IL-6, $TNF-{\alpha}$, $PGE_2$, and $LTB_4$ production amounts were significantly reduced in the EST, SCT, WDT extraction groups compared with the control group, and significantly decreased the amount of $NF-{\kappa}B$, iNOS, and COX-2 gene expression and the amount of Phospho-Inhibitor kappa B alpha ($p-I{\kappa}B-{\alpha}$)/Inhibitor kappa B alpha ($I{\kappa}B-{\alpha}$) and $NF-{\kappa}B$ p65 protein expression. In addition, As a result of measuring the anti-pruritic effect, the amounts of histamine, ACh and Substance P were significantly decreased, and AChE production was slightly decreased, but it's significance did not appear. Finally the anti-microbial effects of EST, SCT, WDT extraction groups against Pseudomonas aeruginosa, Candida albicans and Aspergillus niger was inhibited, however the growth of Escherichia coli and Staphylococcus aureus was not inhibited. Conclusions: These data suggest that EST, SCT, WDT can be used to treat patients with leukorrhea.

Vasodilatory Effects of Samhwangsasim-tang on Vascular Smooth Muscle (삼황사심탕의 혈관이완 효능과 기전)

  • Kim Jong Bong;Kwon Oh Kui;Son Chang Woo;Shin Heung Mook
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.5
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    • pp.1382-1386
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    • 2004
  • This study was performed for the investigation of vasodilatory efficacy and its underlying mechanisms of Samhwangsasim-tang(SST), herbal remedy. SST relaxed vascular strips precontracted with phenylephrine or KCI(51 mM), but the magnitude of relaxation was greater in phenylephrine(PE) induced contraction. The relaxation effects of SST was endothelium-independent. L-NAME, iNOS inhibitor, and methyl en blue(MB), cGMP inhibitor, did not attenuate the relaxation responses of SST. In the absence of extracellular Ca2+, pre-incubation of the aortic rings with SST significantly reduced the contraction by PE, suggesting that the relaxant action of the SST includes inhibition of Ca/sup 2+/ influx and release of Ca/sup 2+/ from intracellular stores (SR). In addition, the cell death was induced by SST in human aortic smooth muscle cells but not that of human umbilical vein endothelial cells. We conclude that in rat thoracic aorta, SST may induce in part vasodilation through inhibition of Ca/sup 2+/ influx and release of Ca/sup 2+/ from intracellular stores.

NSA9, a human prothrombin kringle-2-derived peptide, acts as an inhibitor of kringle-2-induced activation in EOC2 microglia

  • Kim, Ji-Yeon;Kim, Tae-Hyong;Kim, Soung-Soo
    • BMB Reports
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    • v.42 no.6
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    • pp.380-386
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    • 2009
  • In neurodegenerative diseases, such as Alzheimer' and Parkinson', microglial cell activation is thought to contribute to CNS injury by producing neurotoxic compounds. Prothrombin and kringle-2 increase levels of NO and the mRNA expression of iNOS, IL-1$\beta$, and TNF-$\alpha$ in microglial cells. In contrast, the human prothrombin kringle-2 derived peptide NSA9 inhibits NO release and the production of pro-inflammatory cytokines such as IL-1$\beta$, TNF-$\alpha$, and IL-6 in LPS-activated EOC2 microglia. In this study, we investigated the anti-inflammatory effects of NSA9 in human prothrombin- and kringle-2-stimulated EOC2 microglia. Treatment with 20-100 ${\mu}M$ of NSA9 attenuated both prothrombin- and kringle-2-induced microglial activation. NO production induced by MAPKs and NF-$\kappa$B was similarly reduced by inhibitors of ERK (PD98059), p38 (SB203580), NF-$\kappa$B (N-acetylcysteine), and NSA9. These results suggest that NSA9 acts independently as an inhibitor of microglial activation and that its effects in EOC2 microglia are not influenced by the presence of kringle-2.

Effects of Cnidium officinale, Petasites japonicus, Coptis chinensis Extract Mixture on Vasodilation (천궁(川芎), 머위, 황련(黃連) 추출물 조성의 $Ca^{2+}$ 유입 억제를 통한 혈관이완 효능)

  • Kim, Sang-Dae;김길훤, Gil-Whon;Shin, Heung-Mook
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.6
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    • pp.1620-1624
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    • 2006
  • This study was performed to investigate a vasodilatory efficacy and its underlying mechanisms of the mixture of Cnidium officinale, Petasites japonicus and Coptis chinensis (CPC), CPC relaxed rat aortic vascular strips in endothelium-independant manner precontracted with phenylephrine or KCI(50mM), but the magnitude of relaxation was greater in KCI induced contraction. L-NAME, iNOS inhibitor, and methylen blue(MB), cGMP inhibitor, did not attenuate the relaxation responses of CPC. Furthermore, the contraction by increaseing $Ca^{2+}$ concentration (0.3-10.0mM) to a $Ca^{2+}$-free high $K^+$ (60mM) was significantly reduced by CPC pretreatment. These results suggest that the relaxation effect of CPC is related with the block of $Ca^{2+}$ influx via $Ca^{2+}$ channel.

Anti-Inflammatory Effect of Erigeron annuus L. Flower Extract through Heme Oxygenase-1 Induction in RAW264.7 Macrophages (RAW264.7 대식세포에서 Heme Oxygenase-1의 유도에 의한 개망초 (Erigeron annuus L.) 꽃 Methanol 추출물의 항염증 효과)

  • Sung, Mi-Sun;Kim, Young-Hwa;Choi, Young-Min;Ham, Hyeon-Mi;Jeong, Heon-Sang;Lee, Jun-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.11
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    • pp.1507-1511
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    • 2011
  • This study investigated the anti-inflammatory effect of Erigeron annuus L. flower (EAF) methanol extract. We examined the involvement of heme oxygenase-1 (HO-1) in the inhibitory activities of EAF methanol extract on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. Cell viability and NO assays were performed. In addition, inducible nitric oxide synthase (iNOS) and HO-1 expressions were detected by Western blotting and blocking HO-1 activity on NO production. EAF methanol extract (25, 50, 100, 200 ${\mu}g$/mL) significantly inhibited LPS-stimulated NO production (p<0.05; 12.82, 9.61, 6.83, 2.52 ${\mu}m$) in a concentration-dependent manner. EAF methanol extract also reduced the expression of iNOS protein. The EAF methanol extract induced the expression of HO-1 in a dose-dependent manner. Blockage of HO-1 activity by zinc protoporphyrin suppressed EAF methanol extract-induced reductions in the production of NO. The present results suggest that EAF methanol extract has a potent anti-inflammatory effect in RAW264.7 macrophages through the induction of HO-1.

THE EFFECT OF DIFFERENTIAL MODULATION OF N-METHYL-D-ASPARTATE RECEPTOR ON THE PROLIFERATION OF PRIMARY CULTURED NORMAL HUMAN ORAL KERATINOCYTES: DNA SYNTHESIS RATE ANALYSIS (N-methyl-D-aspartate 수용기의 다양한 조절이 일차 배양된 정상사람구강각화세포의 증식에 미치는 영향; DNA 합성율 평가)

  • Kim, In-Soo;Paik, Ki-Suk;Chang, Mi-Sook;Lee, Won;Lee, Seung-Pyo
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.33 no.2
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    • pp.124-130
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    • 2007
  • In the present study, I investigated the effects of N-methyl-D-aspartate (NMDA), arachidonic acid (AA), and Nitric Oxide Synthase Inhibitor (NOSI), alone or in combination, on the proliferation of cultured primary normal human oral keratinocytes (NHOK). The purpose of this study was therefore the preliminary study for the examination of the interaction between these agents and NHOK in order to elucidate the mechanisms by which epithelial growth and regeneration are regulated. NHOK were obtained from gingival tissue of 20 individuals aged 20 to 29, and third passage (P3) cells were used for this study. The DNA synthesis was measured by the BrdU assay. Addition of low concentration of AA ($1{\mu}M$) and high concentration of AA with NMDA group (NMDA+AA $10{\mu}M$) made DNA synthesis rate increase significantly at the early stage. Adding NNA ($10{\mu}M$) affected DNA synthesis rate to increase significantly in 4 hours. At the early stage, DNA synthesis was significantly active in the NOS-I with NMDA groups than in the control and the NMDA-only group, while it didn't become statistically meaningful in 24 hours. AA $1{\mu}M$ and NNA $10{\mu}M$ may induce the proliferation of the NHOK independently and NOS-I may induce the proliferation of the NHOK with NMDA. These reactions might be related to the NMDA receptor in the cell and the change of the intracellular calcium ion concentration.

Synergistic Anti-inflammatory Effect of Rosmarinic Acid and Luteolin in Lipopolysaccharide-Stimulated RAW264.7 Macrophage Cells (Rosmarinic acid와 luteolin의 항염증에 대한 상승효과)

  • Cho, Byoung Ok;Yin, Hong Hua;Fang, Chong Zhou;Ha, Hye Ok;Kim, Sang Jun;Jeong, Seung Il;Jang, Seon Il
    • Korean Journal of Food Science and Technology
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    • v.47 no.1
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    • pp.119-125
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    • 2015
  • The aim of this study was to investigate the synergistic anti-inflammatory effect of rosmarinic acid (RA) and luteolin from perilla (Perilla frutescens L.) leaves in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages. A combination of RA and luteolin more strongly inhibited the production of nitric oxide (NO), inducible NOS (iNOS), prostaglandin $E_2$ ($PGE_2$), and COX-2 than higher concentrations of RA or luteolin alone in LPS-stimulated RAW264.7 macrophages. The combined RA and luteolin synergistically inhibited the production of pro-inflammatory cytokines, such as tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-6 (IL-6), and interleukin-$1{\beta}$ (IL-$1{\beta}$), in LPS-stimulated RAW264.7 macrophages. Furthermore, combined RA and luteolin more strongly suppressed NF-${\kappa}B$ activation than RA or luteolin alone, by inhibiting the degradation of inhibitor of NF-${\kappa}B(I{\kappa}B)$-${\alpha}$ and nuclear translocation of the p65 subunit of NF-${\kappa}B$ in LPS-stimulated RAW264.7 macrophages. Collectively, these results suggest that RA and luteolin in combination exhibit synergistic effects in suppression of LPS-induced inflammation in RAW264.7 macrophages.

Inhibition of LPS-induced NO Production and NT-$\textsc{k}B$ Activation by a Sesquiterpene from Saussurea lappa

  • Jin, Mirim;Lee, Hwa-Jin;Ryu, Jae-Ha;Chung, Kyu-Sun
    • Archives of Pharmacal Research
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    • v.23 no.1
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    • pp.54-58
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    • 2000
  • To elucidate the molecular mechanisms for the suppression of LPS-induced nitric oxide (NO) production by a dehydrocostus lactone (DL) from Saussurea lappa, we examined the preventive effect of this compound on $NF-{\kappa}B$ activation in LPS-treated RAW 264.7 macrophages and U937 human monocytic cells. The results suggest that the suppression of NO production is mediated by the inhibitory action on the i-NOS gene expression through the inactivation of $NF-{\kappa}B$ and this sesquiterpene lactone can act as a pharmacological inhibitor of the $NF-{\kappa}B$ activation.

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Inhibitory Constituents of LPS-induced Nitric Oxide Production from Arctium lappa

  • Park, So-Young;Hong, Seong-Su;Han, Xiang-Hua;Ro, Jai-Seup;Hwang, Bang-Yeon
    • Natural Product Sciences
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    • v.11 no.2
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    • pp.85-88
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    • 2005
  • The methanolic extract from the seeds of Arctium lappa was found to inhibit the LPS-induced nitric oxide (NO) production in murine macrophage RAW264.7 cells. Bioassay-guided fractionation of a methylene chloride soluble fraction led to the isolation of three lignan compounds, arctiin (1), arctigenin (2), and lappaol B (3). Their structures were elucidated by UV, IR, MS, and NMR data, as well as by comparison with those of the literatures. Arctigenin (2) and lappaol B (3) had an iNOS inhibitory activity with $IC_{50}$ values of 12.5 and $25.9\;{\mu}M$, respectively.