• Title/Summary/Keyword: Inducible nitric oxide

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Luteolin and Chicoric Acid, Two Major Constituents of Dandelion Leaf, Inhibit Nitric Oxide and Lipid Peroxide Formation in Lipopolysaccharide-Stimulated RAW 264.7 Cells

  • Park, Chung-Mu;Park, Ji-Young;Song, Young-Sun
    • Preventive Nutrition and Food Science
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    • v.15 no.2
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    • pp.92-97
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    • 2010
  • Luteolin and chicoric acid are the most abundant phytochemicals in dandelion (Taraxacum officinale) leaf. In this study, four kinds of extraction methods [hot water, ambient temperature (AT) water, ethanol, and methanol] were applied to analyze the contents of both phytochemicals and verify their anti-inflammatory and antioxidative activities. The methanol extract showed the most potent nitric oxide (NO) inhibitory effect. The luteolin and chicoric acid concentrations were 3.42 and $12.86\;{\mu}g/g$ dandelion leaf in the methanol extract. The NO-suppressive effect of luteolin and chicoric acid was identified in a dose-dependent manner with $IC_{50}$ values of $21.2\;{\mu}M$ and $283.6\;{\mu}M$, respectively, in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells without cytotoxicity. Malondialdehyde (MDA) concentration, as an index for free radical injury on cell membrane, was also dose-dependently inhibited by the two compounds. The suppressive effect was further examined using mRNA and protein expression levels, which were attributable to the inhibition of inducible nitric oxide synthase (iNOS). These results suggest that two phytochemicals in dandelion leaf, luteolin and chicoric acid, may play an important role in the amelioration of LPS-induced oxidative stress and inflammation.

Inhibitory effect of Yongdamsagantang water extract on IL-6 and nitric oxide production in lipopolysaccharide-activated RAW 264.7 cells

  • Lim, Jin-Ho;Lee, Jong-Rok;Kim, Sang-Chan;Jee, Seon-Young
    • Advances in Traditional Medicine
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    • v.7 no.3
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    • pp.321-329
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    • 2007
  • The present study was conducted to evaluate the effect of Yongdamsagantang (YST) on the regulatory mechanism of cytokines and nitric oxide (NO) for the immunological activities in RAW 264.7 cells. After the treatment of YST water extract, cell viability was measured by MTT assay, and NO production was monitored by measuring the nitrite content in culture medium. Inducible nitric oxide synthase (iNOS) and phospholylation of inhibitor of nuclear factor kappa B alpha ($p-I{\kappa}B{\alpha}$) were determined by Immunoblot analysis, and levels of cytokine were analyzed by sandwich immunoassays. Results provided evidences that YST inhibited the production of NO. iNOS, and interleukin-6, and the activation of $p-I{\kappa}B{\alpha}$ in RAW 264.7 cells activated with lipopolysaccharide. These findings showed that YST could have some anti-inflammatory effects which might play a role in therapy in Gram-negative bacterial infections.

Raloxifene, a Selective Estrogen Receptor Modulator, Inhibits Lipopolysaccharide-induced Nitric Oxide Production by Inhibiting the Phosphatidylinositol 3-Kinase/Akt/Nuclear Factor-kappa B Pathway in RAW264.7 Macrophage Cells

  • Lee, Sin-Ae;Park, Seok Hee;Kim, Byung-Chul
    • Molecules and Cells
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    • v.26 no.1
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    • pp.48-52
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    • 2008
  • We here demonstrate an anti-inflammatory action of raloxifene, a selective estrogen receptor modulator, in lipopolysaccharide (LPS)-induced murine macrophage RAW264.7 cells. Treatment with raloxifene at micromolar concentrations suppressed the production of nitric oxide (NO) by down-regulating expression of the inducible nitric oxide synthase (iNOS) gene in LPS-activated cells. The decreased expression of iNOS and subsequent reduction of NO were due to inhibition of nuclear translocation of transcription factor NF-${\kappa}B$. These effects were significantly inhibited by exposure to the phosphatidylinositol 3-kinase (PI 3-kinase) inhibitor, LY294002, or by expression of a dominant negative mutant of PI 3-kinase. In addition, pretreatment with raloxifene reduced LPS-induced Akt phosphorylation as well as NF-${\kappa}B$ DNA binding activity and NF-${\kappa}B$-dependent reporter gene activity. Thus our findings indicate that raloxifene exerts its anti-inflammatory action in LPS-stimulated macrophages by blocking the PI 3-kinase-Akt-NF-${\kappa}B$ signaling cascade, and eventually reduces expression of pro-inflammatory genes such as iNOS.

Effect of Immunosuppressants on Lipopolysaccharide-Induced Changes of Nitric Oxide Synthase Activity in Liver and Brain of Mice (면역억제제가 Lipopolysaccharide에 의한 생쥐의 간 및 뇌조직의 Nitric Oxide Synthase 활성도의 변화에 미치는 영향)

  • Min, Byung-Woo;Han, Hyng-Soo;Park, Jung-Sook;Kim, Choong-Young
    • The Korean Journal of Pharmacology
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    • v.31 no.2
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    • pp.233-239
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    • 1995
  • To verify the effect of immunosuppressants on the endotoxin-induced increase in iNOS activity, the action of immunosuppressants, dexamethasone (1.5 mg/kg), azathioprine (5 mg/kg/day) and cyclosporine (10 mg/kg), were evaluated in mice pretreated with LPS. The intraperitoneal injection of lipopolysaccharide (10 mg/kg) increased the nitric oxide synthase (NOS) activity in the brain and liver to maximum at 1 and 3 hours, respectively. The increase in NOS activity was blocked by the treatment with NOS inhibitor, LNAME(300 mg/kg) and aminoguanidine(100 mg/kg); a protein inhibitor, cycloheximide (10 mg/kg); and a transcription inhibitor of inducible NOS(iNOS), dexamethasone(1.5 mg/kg). Immunosuppressants, azathioprine (5 mg/kg) and cyclosporine (10 mg/kg), effectively blocked the increase in NOS activity. These results suggest that iNOS expression plays an important role in LPS-induced the increase in NOS activity and that immunosuppressants can be used as candidate for therapeutic agents in endotoxemia.

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Effects of NOS Inhibitors on Arthritis and Arthritic Pain in Rats

  • Min, Sun-Seek
    • The Korean Journal of Physiology and Pharmacology
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    • v.11 no.6
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    • pp.253-257
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    • 2007
  • Among the arthritis symptoms, chronic pain is the most serious, and it can profoundly affect the quality of human life. Unfortunately, the mechanism of development in arthritis and arthritic pain has not yet been precisely elucidated. Accumulating evidence indicates that nitric oxide (NO) plays a pivotal role in nociceptive processing in the spinal cord. However, the modulation mechanism of NO in the peripheral site of arthritis and arthritic pain has not been clarified. Therefore, I determined in the present study which nitric oxide synthase (NOS) was involved in the induction of arthritis and arthritic pain. Monoarthritis was induced by intra-articular injection of carrageenan (2%, $50{\mu}l$) into rats, and resulted in the reduction of weight load on the injected leg, increase of knee joint diameter and inflammatory response. Pre-treatment of rats with L-N6-(1-iminoethyl)-lysine (L-NIL, $500{\mu}g$, in $50{\mu}l$), an inhibitor of inducible NOS (iNOS), partially prevented the induction of pain-related behavior and partially reduced inflammatory response in the synovial membrane in the knee joint. These results suggest that iNOS in the knee joint may play an important role in the induction of pain-related behavior and inflammation, and that NO produced by iNOS may be associated with nociceptive signaling in the peripheral site.

Inhibitory Effects of Aralia cordata Thunb Extracts on Nitric Oxide Synthesis in RAW 264.7 Macrophage Cells (독활(Aralia cordata Thunb) 추출물의 Nitric Oxide Synthesis 저해효과)

  • Kang, Chang-Ho;Koo, Ja-Ryong;So, Jae-Seong
    • Korean Journal of Food Science and Technology
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    • v.44 no.5
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    • pp.621-627
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    • 2012
  • Assessment was made of the effects of Aralia cordata Thunb (DH) on the cell proliferation, inducible nitric oxide synthase (iNOS) mRNA gene expression and nitric oxide (NO) production in RAW 264.7 macrophage cells. For the screening of anti-inflammatory activities, ethanolic extracts of 55 species of traditional herbal medicines were examined for inhibitory effects, and it was confirmed that DH possessed inhibitory effects on NO production. As a result, DH significantly decreased the production of NO and iNOS gene expression at a concentration of $250{\mu}g/mL$. The chloroformsoluble fractionates have the strongest No synthesis inhibitory effect. It is presumed that the inhibition of NO production in LPS-stimulated RAW 264.7 cells by DH components occurred via the modulation of iNOS and DH, and that the active compound from DH may be useful for therapeutic management of inflammatory-associate diseases.

Inhibitory Effect of Chan-Su on the Secretion of PGE2 and NO in LPS-stimulated BV2 Microglial Cells

  • Kim, Min-Hee;Lyu, Ji-Hyo;Lyu, Sun-Ae;Hong, Sang-Hoon;Kim, Won-Il;Yoon, Hwa-Jung;Ko, Woo-Shin
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.5
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    • pp.1315-1321
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    • 2008
  • Chan-Su (Venenum bufonis) has long been for a variety of other purposes including treatment of inflammation in the folk medicine recipe. Since nitric oxide (NO) is one of the major inflammatory parameters, we first studied the effects of Chan-Su on NO production in lipopolysaccharide (LPS)-stimulated BV2 microglial cells, Chan-Su inhibited the secretion of NO in BV2 microglial cells, without affecting cell viability, The protein level of inducible nitric oxide synthase (iNOS) was decreased by Chan-Su, And Chan-Su also inhibited production of prostaglandin E2 (PGE2) and expression of cyclooxygenase (COX)-2. Proinflammatory cytokines, such as tumor necrosis factor $(TNF)-{\alpha}$, interleukin $(IL)-1{\beta}$ and IL-12, were inhibited by Chan-Su in a dose-dependent manner. And Chan-Su inhibited the degradation of ${IkB-\alpha}$, which was considered to be inhibitor of nuclear factor $(NF)-{\kappa}B$, one of a potential transcription factor for the expression of iNOS, COX-2 and proinflammatory cytokines. These results suggest that Chan-Su could exert its anti-inflammatory actions by suppressing the synthesis of NO through inhibition of $I{\kappa}B-{\alpha}$ degradation.

Antioxidant and Anti-inflammatory Effects of Bletilla striata Reichenbach fil. Fractions as Cosmetic (백급(Bletilla striata Reichenbach fil.) 분획물의 항산화, 항염증 효과와 화장품소재로서의 연구)

  • Yoon, Ji-Hun;Park, Seong-Geun;Lee, Mi-Ji;Park, Jin-Young;Seo, Kyo-Seong;Woo, Kyung-Chul;Lee, Chang-Eon
    • Journal of Life Science
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    • v.23 no.9
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    • pp.1073-1078
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    • 2013
  • The antioxidant and anti-inflammatory effects of active ingredients in cosmetics are very important. The effects are closely related to the prevention of skin aging. Among medicinal plants, Bletilla striata Reichenbach fil. has well-known pharmacological activity. Extracted samples were prepared using sequential fractionation of ethyl acetate, butanol, and water. The antioxidant effect of the fractions was confirmed by DPPH and ABTS+ radical scavenging. Among the various fractions of B. striata Reichenbach fil., ethyl acetate was associated with a reduction in nitric oxide production, which was induced by LPS (lipopolysaccharide) treatment in a dose-dependent manner. In addition, the production of iNOS (inducible nitric oxide synthase) and COX-2 (cyclooxygenase-2), which are upstream regulators of nitric oxide production, was also inhibited. Thus, the ethyl acetate fraction of B. striata Reichenbach fil. appears to be a potential active ingredient for use in cosmetics.

Inhibition of Nitric Oxide Production by Coumarins from Peucedanum japonicum in LPS-Activated RAW 264.7 cells (갯기름나물의 쿠마린에 의한 RAW 264.7 세포주의 Nitric Oxide 생성 저해활성)

  • Choi, Hee-Cheol;Rho, Tae-Cheol;Kim, Bo-Yeon;Ko, Hack-Ryong;Oh, Won-Keun;Seong, Chang-Keun;Mheen, Tae-Ick;Ahn, Jong-Seog;Lee, Hyun-Sun
    • Korean Journal of Pharmacognosy
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    • v.30 no.2
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    • pp.99-104
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    • 1999
  • During the screening for inhibitors of nitric oxide production in LPS-activated macrophage, RAW 264.7 cells. Five coumarins were isolated from chloroform extract of the root of Peucedanum japonicum. They were identified as praeruptorin A (1), xanthotoxin (2), psoralen (3), isopimpinellin (4), bergapten (5) on the basis of spectroscopic methods. The $IC_{50}$ values for nitrite production by activated macrophages were about $1.5\;{\mu}g/ml$ (1), $0.3\;{\mu}g/ml$ (2), $1.0\;{\mu}g/ml$ (3), $25\;{\mu}g/ml$ (4), $25\;{\mu}g/ml$ (5), respectively. However, the inducible nitric oxide synthase (iNOS) was not inhibited by treatment with these compounds. Their inhibitory effect on nitric oxide production was resulted from the supperssion of iNOS expression.

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Inhibitory effect of Chelidonii Herba water extract on production of Nitric Oxide, Expression of iNOS and COX-2 in lipopolysaccharide-activated Raw 264.7 cells (백굴채(白屈菜)의 물추출물이 lipopolysaccharide로 유도된 Nitric Oxide의 생성 및 iNOS와 COX-2의 발현에 미치는 영향)

  • Zhao Rong-Jie;Kim Young-Woo;Byun Sung-Hui;Kim Sang-Chan
    • Herbal Formula Science
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    • v.12 no.2
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    • pp.163-173
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    • 2004
  • Chelidonii Herba (CHE, Baek-gul-chae in Korean), which has its original description in Gu-Hwang-Bon-Cho, a classic book of oriental Herbal book, is widely used in the treatment of stomach cancer, jaundice, gasrtic ulcer, edema and stomach pain, in Korea, Japan and China. The present study was conducted to evaluate the effect of CHE on the nitric oxide (NO) production, iNOS and COX-2 expression in lipopolysaccharide - activated Raw 264.7 cells. After the treatment of CHE, NO production was monitored by measuring the nitrite content in culture medium, cell viability was measured by MIT assay. COX-2 and iNOS were determined by lmmunoblot analysis. The production of nitric oxide was significantly inhibited by pretreatment (1h) with CHE (0.1-0.3 mg/ml) on LPS-activated Raw264.7 cells. The expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) protein were up-regulated by LPS, but the increased levels of iNOS and COX-2 were inhibited by pretreatment of CHE (0.1-0.3 mg/ml), respectively. Thus, the present data suggest that CHE may play an important role in adjunctive therapy in Gram-negative bacterial infections.

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