• Title/Summary/Keyword: nitric oxide synthase activity

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Antioxidant and Anti-inflammatory Effect of Angelica Tenuissima in IFN-${\gamma}$/LPS-stimulated Peritoneal Macrophage (IFN-${\gamma}$와 LPS로 자극된 쥐의 복강 대식세포에서 고본(藁本) 메탄올 추출물의 항염증 효과)

  • Lee, Se-Yeoun;Park, Ho-Jun;Cha, Dong-Seok;Shin, Tae-Yong;Na, Ho-Jeong;Moon, Woo-Sung;Kang, Yang-Gyu;Jeon, Hoon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.6
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    • pp.1549-1556
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    • 2008
  • The objective of this study were to evaluate the antioxidant activity and the anti-inflammatory effects of Angelica tenuissima (AT) which has been used widely as a traditional medicine. The antioxidant activities of AT was tested by DPPH radical scavenging, superoxide anion scavenging and nitric oxide scavenging. AT showed strong antioxidant activity in all experiment. In macrophages nitric oxide (NO) is released as an inflammatory mediator and has been proposed to be an important modulator of many pathophysiological conditions and high concentratin of NO is produced by inducible nitric oxide synthase (iNOS). In this study we have examined the inhibition effects of NO by 85% methanol extracts of AT in mouse (C57BL/6) peritoneal macrophage. AT (100, 1000 ${\mu}g/m{\ell}$) suppressed nitric oxide production and iNOS expression without any notable cytotoxicity and it also inhibited the expression of inflammatoryenzymes like cyclooxygenase-2 (COX-2). These data suggest that 85% methanol extracts of AT may possibly be used as antioxidant and anti-inflammatory agent.

In vitro anti-inflammatory activity of extracts from Potentilla supina in murine macrophage RAW 264.7 cells (개소시랑개비 추출물의 RAW264.7대식세포에서 in vitro 항염효과)

  • Nam, Jung-Hwan;Kim, Hyun-Sam;Kim, Byoumg-Jin;Yu, Hong-Seob;Chang, Dong-Chil;Jin, Yong-Ik;Yoo, Dong-Lim;Choi, Jong-Keun;Park, Hee-Jhun;Lee, Seung-Bin;Lee, Kyung-Tea;Park, Soo-Jin
    • Journal of Plant Biotechnology
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    • v.44 no.1
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    • pp.76-81
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    • 2017
  • Potentilla supina (Rosaceae) has traditionally been used to treat disorders of hemostasis, dysentery, malaria, bloody discharge and arthritis, and it has antinociceptive and anti-inflammatory properties. However, validity of the anti-inflammatory activity has not been scientifically investigated so far. Therefore, the aim of this study was to investigate the anti-inflammatory potential of P. supina using the ethanolic extract of P. supina and its sub-fractions. To evaluate the anti-inflammatory effects of P. supina, we examined the inflammatory mediators such as nitric oxide (NO), inducible nitric oxide synthase (iNOS) and prostaglandin $E_2$ ($PGE_2$) in RAW 264.7 cells. Our results indicated that ethyl acetate fraction significantly inhibited LPS-induced NO, iNOS and $PGE_2$ production in RAW 264.7 cells. This result showed that ethyl acetate fraction of P. supina is expected to be a good candidate for development into a source of anti-inflammatory agents.

Inhibitory Effect of Taraxci Herba Methanol Extract on Pro-inflammatory Mediator in Lipopolysaccharide;Activated Raw 264.7 cells

  • Jo, Mi-Jeong;Chu, Yan-Hui;Back, Young-Doo;Lee, Byung-Wook;Shin, Soon-Shik;Kwon, Young-Kyu;Kim, Sang-Chan
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.4
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    • pp.907-913
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    • 2008
  • Taraxci Herba (TH; Pogongyoung in Korean) has been used in traditional oriental medicine for the treatment of various ailments. The biological activity of this plant is not yet evaluated systematically. This study was conducted to evaluate the inhibitory effects of TH on the production of nitric oxide (NO) and the expression of inducible nitric oxide synthase (iNOS) in lipopolysaccharide (LPS)-activated Raw264.7 cells. The aim of the present work is to investigate a potential anti-inflammatory activity of TH. The Raw264.7 cells were cultured in DMEM medium for 24 h. After serum starvation for 12 h, the cells were treated with TH for 1 h, followed by stimulating NO production with LPS ($2{\mu}g/ml$). As result of this study, TH inhibited the levels of NO, PGE2, $TNF-{\alpha}$, IL-6 and $IL-1{\beta}$, and the expressions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) activated by LPS. These inhibitory effects were mediated though the inhibition of phosphorylation of inhibitory kappa B ($I{\kappa}B$). These findings showed that TH could have some anti-inflammatory effects.

m2 Muscarinic Receptors Stimulate Neuronal Nitric Oxide Synthase

  • Lee, Seok-Yong;Park, Sun-Hye;Cho, Tai-Soon
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.208-208
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    • 1996
  • In this work we investigated coupling of the m2 and m4 subtypes of muscarinic acetylcholine receptors expressed in chinese hamster ovary (CHO) cells to activation of neuronal nitric oxide synthase (nNOS). Stimulation of guanylate cyclase activity in detector neuroblastoma cells was used as an index of generation of nitric oxide (NO) in CHO cells. The agonist carbachol induced marked time and concentration-dependent enhancement of the activity of nNOS at m2 receptors. In sharp contrast, the response in CHO cells transfected with the m4 receptor gene was similar in magnitude to that observed in non-transfected cells, suggesting lack of significant coupling of m4 muscarinic receptors to NO signaling. This novel observation of functional divergence of the two muscarinic receptor subtypes at the level of activation of nNOS is quite intriguing, in light of the currently accepted dogma that they belong to the same functional class. This functional selectivity was not due to differential effects on intracellular Ca$\^$2+/ concentration, since activation of both subtypes of muscarinic receptors produced a comparable, albeit quite small, Ca$\^$2+/ signal. Taken together, our present data strongly suggest that the generally assumed functional equivalence of m2 and m4 muscarinic receptors should be carefully reexamined. These data also suggest the presence of alternate mechanisms of activation of nNOS, which might be operative in the absence of large changes in the concentration of cellular Ca$\^$2+/. The latter mechanisms are expected to be activated by m2, but not m4 muscarinic receptors. Both sets of findings are quits important in regards to refining the functional classification of muscarinic receptor subtypes and the cellular mechanisms of activation of NOS.

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The Inhibitory Effect of Lycii Fructus on LPS-stimulated NF-${\kappa}B$ Activation and iNOS Expression in RAW 264.7 Macrophages

  • Kim, Beum-Seuk;Song, Yun-Kyung;Lim, Hyung-Ho
    • The Journal of Korean Medicine
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    • v.29 no.1
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    • pp.47-59
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    • 2008
  • Objective : Anti-inflammatory effects of the extract of Lycii Fructus on lipopolysaccharide (LPS)-induced inflammation in RAW 264.7 macrophage cells were investigated. Method : In order to assess the cytotoxic effect of Lycii Fructus on the raw 264.7 macrophages 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay was performed. Reverse transcription-polymerase chain reaction(RT-PCR) analysis of the mRNA levels of tumor necrosis factor-$\alpha$(TNF-$\alpha$) and inducible nitric oxide synthase(iNOS) was performed in order to provide an estimate of the relative level of expression of these genes. The protein level of the inhibitor of nuclear factor-${\kappa}B(I{\kappa}B)$ and nuclear factor-${\kappa}B$(NF-${\kappa}B$) activity was investigated by Western blot assay. NO production was investigated by NO detection. Result : Lycii Fructus suppressed NO production by inhibiting the LPS-induced expressions of iNOS and TNF-$^-\alpha$ mRNA and iNOS protein in RAW 264.7 macrophage cells. Also, Lycii Fructus suppressed activation of NF-${\kappa}B$ in the nucleus. Conclusion : These results show that the extract of Lycii Fructus has anti-inflammatory effect probably by suppressing iNOS expressions through the down-regulation of NF-${\kappa}B$ binding activity.

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Expression of Nitric Oxide Synthase and Endothelin-1 in Human Uterine Artery from Full-Term Pregnancies

  • Choi, Ook-Hwan;Lee, Sun-Hee;Kim, Eun-Jin;Kim, Koan-Hoi;Rhim, Byung-Yong
    • The Korean Journal of Physiology and Pharmacology
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    • v.9 no.3
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    • pp.165-172
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    • 2005
  • The aim of this study was to determine the roles of ET-1 and NO on uterine blood flow in pregnancy. Uterine arteries were isolated from 17 nonpregnant and 12 pregnant women. Nonpregnant group included patients with median age of $48.6{\pm}2.3$ years who underwent hysterectomy, because of myoma. Pregnant group included patients with median age of $31.3{\pm}1.4$ years undergoing cesarean delivery. ET-1 and ET-2 induced concentration-dependent contraction in isolated nonpregnant and pregnant uterine arteries. The contractile response and maximal contraction were increased in pregnant uterine arteries. In nonpregnant uterine arteries, there was no contraction in response to ET-3, whereas pregnancy induced concentration-dependent contraction by ET-3. Tissue nitrite/nitrate level and immunohistochemical staining of eNOS and iNOS were increased in pregnant uterine arteries, compared with nonpregnant uterine arteries. In addition, the expressions of eNOS and iNOS mRNA were significantly increased in pregnancy. Moreover, contractions by ET isopeptides, including ET-1, were enhanced, and immunohistochemical staining of ET-1 and ET-1 mRNA expression was increased in pregnant uterine arteries. These results suggest that NO production by increased NOS activity, especially eNOS activity, is related to placental and uterine blood flow. Furthermore, ET-1 appears to play a pathophysiological role in pregnant complications such as hypertension.

Lonicera Japonioa Inhibits the Production of NO through the Suppression of NF-kB Activity in LPS-stimulated Mouse Peritoneal Macrophages

  • Kim Young-hee;Kim Han-do
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.17 no.1
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    • pp.163-171
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    • 2004
  • The flowers of Lonicera japonica Thunb. (Caprifoliaceae) has been used as anti-inflammatory drug in the folk medicine recipe and been proved its anti-inflammatory effect in the oriental medicine. However, the action mechanism of Lonicera japonica that exhibits anti-inflammatory effects has not been determined. Since nitric oxide (NO) is one of the major inflammatory parameter, we studied the effect of aqueous extracts of Lonicera japonica (AELJ) on NO production in lipopolysaccharide (LPS)-stimulated mouse peritoneal macrophages. NO and inducible NO synthase (iNOS) level were significantly reduced in LPS-stimulated macrophages by AELJ compared to those without Electrophoretic mobility shift assay (EMSA) indicated that AELJ blocked the activation of nuclear factor kappa B (NF-kB), which was considered to be a potential transcription factor for the iNOS expression. AELJ also blocked the phosphorylation and degradation of inhibitor of kappa B-alpha (IkB-${\alpha}$). Furthermore, IkB kinase alpha (IKK${\alpha}$), which is known to phosphorylate serine residues of IkB directly, is inhibited by AELJ in vivo and in vitro. These results suggest that AELJ could exert its anti-inflammatory actions by suppressing the synthesis of NO through inhibition of NF-kB activity.

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Protective Effects of Chlorella vulgaris Extract on Carbon Tetrachloride-induced Acute Liver Injury in Mice

  • Kim, Hyun-Kyung;Li, Li;Lee, Hyeong-Seon;Park, Mi-Ok;Bileha, Dinesh;Li, Wei;Kim, Yong-Ho
    • Food Science and Biotechnology
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    • v.18 no.5
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    • pp.1186-1192
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    • 2009
  • The purpose of this study was to evaluate the protective effects of Chlorella vulgaris extract (CVE) against carbon tetrachloride ($CCl_4$)-induced hepatotoxicity in mice. The mice received silymarin (100 mg/kg), intragastrieally (i.g.) and CVE (50, 100, and 200 mg/kg, i.g.), respectively, every other day, for 4 weeks before $CCl_4$ administration. Twenty-four hr after the administration of $CCl_4$, the serum and liver were analyzed. Our study found that in the CVE groups, aspartate aminotransferase (AST), and alanine aminotransferase (ALT) levels had decreased significantly and the tissue injury was notably diminished compared to the $CCl_4$ group. The antioxidant activities of CVE groups, such as superoxide dismutase (SOD), catalase, and glutathione (GSH), were significantly increased and the activity of nitric oxide synthase (NOS) was remarkably increased in a CVE concentration-dependent manner. In the CVE groups, cytochrome P450 2B1/2B2 (CYP2B1/2) content was decreased. These results indicate that CVE has protective effects against $CCl_4$-induced hepatotoxicity via stimulation of the antioxidant activity and nitric oxide (NO) production, and through inhibition of CYP2B1/2.

Study on the effects of herbal extracts on tooth whitening, antioxidant, nitric oxide synthesis and Streptoccocus mutans. (數種 韓藥材의 齒牙美白 效果에 관한 實驗的 硏究)

  • Jeong, Hyun-A;Rho, Seok-Sun;Lim, Seok-Yin
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.15 no.2
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    • pp.1-19
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    • 2002
  • Fifteen herbs used in oral medicine were extracted in ethanol and screened for tooth whitening effect, radical scavenging activity, inhibition of inducible nitric oxide synthase activity and anti-bacterial activity on Streptoccocus mutans. The results are as followed: 1. The tooth whitening effect of toothpaste containing 1 $\%$ ethanol extracts of 15 herbs was tested in brushing method using artificially stained HAT tablet and bovine tooth specimens. Toothpaste containing 1$\%$ of Vucia unijuga, Illicium verum, Angelica dahurica and Piper longum extracts showed tooth whitening effects on both HAT tablet and bovine tooth specimens. But the others did not show a considerable activity. 2. The antioxidant activity of ethanol extracts of 15 herbs was tested using the method of 1,1-diphenyl-2-picryl hydrazyl (DPPH) reactivity. Two ethanol extracts of Cimicifuga heracleifolia and Phyllostachys nigra var. henonis were found to be the most effective on radical scavenging activity. C. heracleifolia and P. nigra var. henonis extracts removed 86$\%$ and 81 $\%$ of DPPH radical at 0.01$\%$, compared with butylated hydroxy toluene as positive control. 3. The inhibition activity of ethanol extracts on nitrate production In RAW264.7 cell stimulated by lipopolysacchride was tested using the Griess reagent. But all extracts did not inhibit nitrate production. 4. The antibacterial activity on Streptoccocus mutans was tested by paper disk method. But no one extract showed any anti - bacterial activity.

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Thymosin Beta-4, Actin-Sequestering Protein Regulates Vascular Endothelial Growth Factor Expression via Hypoxia-Inducible Nitric Oxide Production in HeLa Cervical Cancer Cells

  • Ryu, Yun-Kyoung;Lee, Jae-Wook;Moon, Eun-Yi
    • Biomolecules & Therapeutics
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    • v.23 no.1
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    • pp.19-25
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    • 2015
  • Vascular endothelial growth factor (VEGF) is an important regulator of neovascularization. Hypoxia inducible nitric oxide (NO) enhanced the expression of VEGF and thymosin beta-4 ($T{\beta}4$), actin sequestering protein. Here, we investigated whether NO-mediated VEGF expression could be regulated by $T{\beta}4$ expression in HeLa cervical cancer cells. Hypoxia inducible NO production and VEGF expression were reduced by small interference (si) RNA of $T{\beta}4$. Hypoxia response element (HRE)-luciferase activity and VEGF expression were increased by the treatment with N-(${\beta}$-D-Glucopyranosyl)-N2-acetyl-S-nitroso-D, L-penicillaminamide (SNAP-1), to generate NO, which was inhibited by the inhibition of $T{\beta}4$ expression with $T{\beta}4$-siRNA. In hypoxic condition, HRE-luciferase activity and VEGF expression were inhibited by the treatment with $N^G$-monomethyl-L-arginine (L-NMMA), an inhibitor to nitric oxide synthase (NOS), which is accompanied with a decrease in $T{\beta}4$ expression. VEGF expression inhibited by L-NMMA treatment was restored by the transfection with pCMV-$T{\beta}4$ plasmids for $T{\beta}4$ overexpression. Taken together, these results suggest that $T{\beta}4$ could be a regulator for the expression of VEGF via the maintenance of NOS activity.