• Title/Summary/Keyword: iNOS, TNF-$\alpha$

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Inhibitory effects of Coptidis Rhizoma on the LPS-induced production of nitric oxide and $TNF-{\alpha}$ in mouse macrophage cells (황련의 쥐 대식세포로부터 LPS에 의해 유도되는 nitric oxide 및 $TNF-{\alpha}$의 생성억제효과)

  • Jung, Hyo-Won;Park, Yong-Ki
    • The Korea Journal of Herbology
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    • v.21 no.2
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    • pp.165-173
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    • 2006
  • Objectives : Coptidis Rhizoma has been known traditional medicine with antimicrobial activities. We investigated inhibitory effects of Coptidis Rhizoma extract on lipopolysaccharide(LPS)-induced nitric oxide production from mouse macrophages. Methods : After Coptidis Rhizoma extract was pretreated in BV2, mouse brain macrophages and RAW264.7 mouse macrophages, cells were activated with LPS. To investigate cytotoxicity Coptidis Rhizoma extract, cell viability was measured by MTT assay. The production of nitric oxide(NO) and inducible nitric oxide synthase(iNOS) was determined in each culture supernatant and mRNA by Griess reaction and RT-PCR. The production of $TNF-{\alpha}$ from cells was measured by ELISA. Results : Coptidis Rhizoma extract significantly inhibited LPS-induced NO production in BV2 and RAW264.7 cells. Coptidis Rhizoma extract also greatly suppressed mRNA expression of iNOS in BV2 and RAW264.7 cells activated by LPS. Conclusion : These data suggests that Coptidis Rhizoma extract may have an anti-inflammatory effect through the inhibition of NO production.

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Immunostimulating activity of maysin isolated from corn silk in murine RAW 264.7 macrophages

  • Lee, Jisun;Kim, Sun-Lim;Lee, Seul;Chung, Mi Ja;Park, Yong Il
    • BMB Reports
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    • v.47 no.7
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    • pp.382-387
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    • 2014
  • Corn silk (CS) has long been consumed as a traditional herb in Korea. Maysin is a major flavonoid of CS. The effects of maysin on macrophage activation were evaluated, using the murine macrophage RAW 264.7 cells. Maysin was isolated from CS by methanol extraction, and preparative $C_{18}$ reverse phase column chromatography. Maysin was nontoxic up to $100{\mu}g/ml$, and dose-dependently increased TNF-${\alpha}$ secretion and iNOS production by 11.2- and 4.2-fold, respectively, compared to untreated control. The activation and subsequent nuclear translocation of NF-${\kappa}B$ was substantially enhanced upon treatment with maysin ($1-100{\mu}g/ml$). Maysin also stimulated the phosphorylation of Akt and MAPKs (ERK, JNK). These results indicated that maysin activates macrophages to secrete TNF-${\alpha}$ and induce iNOS expression, via the activation of the Akt, NF-${\kappa}B$ and MAPKs signaling pathways. These results suggest for the first time that maysin can be a new immunomodulator, enhancing the early innate immunity.

The Effects of Samhwangseje on Anti-Inflammation in Raw 264.7 cell (삼황세제(三黃洗劑)가 Raw 264.7 cell 에서의 항염 효과에 미치는 영향)

  • Jung, Soon-Young;Seo, Hyeong-Sik
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.22 no.1
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    • pp.108-119
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    • 2009
  • Objectives : This study was carried out to investigate the effects of Samhwangseje(SHSJ) on anti-Inflammation in Raw 264.7 cell. Methods : The effects of SHSJ on anti-Inflammation were measured by the cytotoxicity of Raw 264.7 cell, the inhibition for NO, TNF-$\alpha$, $PGE_{2}$, iNOS and COX-2, the blocking NF-${\kappa}B$ into nucleus. Results : 1. All concentrations of SHSJ had no cytotoxicity in Raw 264.7 cell. 2. All concentrations of SHSJ inhibited the production of NO in the Raw 264.7 cell stimulated with LPS. 3. All concentrations of SHSJ did not inhibit the production of TNF-$\alpha$ in the Raw 264.7 cell stimulated with LPS. 4. All concentrations of SHSJ inhibited the production of $PGE_{2}$ in the Raw 264.7 cell stimulated with LPS. 5. All concentrations of SHSJ did not inhibit the expression of COX-2 but concentrations of 50 ${\mu}g/ml$, 100 ${\mu}g/ml$ SHSJ inhibited iNOS expression in the Raw 264.7 cell stimulated with LPS. 6. Concentrations of 50 ${\mu}g/ml$, 100 ${\mu}g/ml$ SHSJ had the effect of blocking NF-${\kappa}B$ into nucleus in LPS-induced macrophage Raw 264.7 cell.

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Anti-inflammatory Effect Of Extracts from Cheongmoknosang(Morus alba L.) in Lipopolysaccharide-stimulated Raw Cells (청목노상 (Morus alba L.)추출물에 의한 Lipopolysaccharide로 유도된 Raw 246.7 cell에서 염증 억제효과)

  • Cho, Young-Je;An, Bong-Jeun
    • Applied Biological Chemistry
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    • v.51 no.1
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    • pp.44-48
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    • 2008
  • With extracts from Cheongmoknosang, anti-inflammatory effect was examined in LPS-stimulated Raw 264.7 cells. LPS (10 ng/ml) treatment increased the production of inflammatory cytokines, $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ but the ethanol extracts from Cheongmoknosang slightly decreased the production of $TNF-{\alpha}$ and also reduced the expression of iNOS and the production of COX-2. It seems that anti-inflammatory effects of ethanol extracts from Cheongmoknosang is partly due to the inhibition of iNOS and COX-2 expression by inhibiting nuclear translocation of $NF-{\kappa}B$ and AP-l in Raw 264.7 cells.

Studies on the NO Production and Expression Induction Effect of NOS Gene by Salviae Radix (단삼에 의한 NO 생성 및 NOS 유전자의 발현 유도 효과에 관한 연구)

  • 조현주;원진희;문구;문석재;유기원;유봉하
    • The Journal of Korean Medicine
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    • v.21 no.3
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    • pp.20-30
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    • 2000
  • Objective : This experiment was performed in order to study the effect of an aqueous extract of Salviae radix root(SRRAE) on NO production and NOS gene induction from macrophages Methods : To investigate dose-dependent effects of SRRAE for NO release on the $rIFN-{\gamma}-treated$ macrophages, the cells were incubated for 6 hrs in a medium containing $rIFN-{\gamma}$ (5 U/ml), stimulated with SRRAE and incubated in a CO2 incubator. The cells were treated with 5 U/ml $rIFN-{\gamma}$ plus 100 g/ml of SRRAE, Then, the cells were incubated with various concentrations of NGMMA at $37^{\circ}C$ for 48 hrs, Results : SRRAE had no effect on NO production by itself, whereas recombinant $interferon-{\gamma}(rIFN-{\gamma})$ alone showed modest activity, When SRRAE was used in combination with $rIFN-{\gamma}$, there was a marked cooperative induction of NO production in a dose-dependent manner. The optimal effect of SRRAE on NO production was shown at 6hrs after treatment with $rIFN-{\gamma}$. The SRRAE-induced production of NO was inhibited by NG-monomethyl- L-arginine(NGMMA) and arginase. $rIFN-{\gamma}$ in combination with SRRAE showed a marked increase of the expression of the inducible NOS(iNOS) gene. In addition, the effect of SRRAE was mainly dependent on the SRRAE-induced tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ secretion. Conclusions : SRRAE induces NO production from macrophages as a result of SRRAE-induced $TNF-{\alpha}$ secretion. SRRAE may provide a second signal for synergistic induction of NO production in macrophages already induced to express iNOS gene by $rIFN-{\gamma}$.

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Inhibition of nitric oxide and TNF-$\alpha$ production by propenone compound through blockaded of NF-$\kappa$B activation in cultured murine macrophages

  • Ju, Hye-Kyung;Lee, Eun-Kyung;Jahng, Yurng-Dong;Lee, Eung-Seok;Chang, Hyeun-Wook
    • Proceedings of the PSK Conference
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    • 2003.10b
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    • pp.156.2-157
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    • 2003
  • Lipopolysaccharide (LPS)-stimulated macrophages produced a large amounts of nitric oxide (NO) by inducible nitric oxide synthase (iNOS). This is an important mechanism in macrophages-induced septic shock and inflammation. In the present study, we tested a synthetic propenone compound, l-furan-2-yl-3-pyridin-2-yl-propenone (FPP-3) for its ability to inhibit the production of tumor necrosis factor-a (TNF-$\alpha$) and an inducible enzyme, iNOS, in the LPS-stimulated murine macrophage-like cell line, Raw264.7. FPP-3 consistently inhibited nitric oxide (NO) and TNF-$\alpha$ production in a dose dependent manner, with $IC_50$> values of 10.0 and 13.1 $\mu$M, respectively. (omitted)

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Inhibitory Effects of Eutigosides Isolated from Eurya emarginata on the Inflammatory Mediators in RAW264.7 Cells

  • Park Soo-Yeong;Lee Hye-Ja;Yoon Weon-Jong;Kang Gyoung-Jin;Moon Ji-Young;Lee Nam-Ho;Kim Se-Jae;Kang Hee-Kyoung;Yoo Eun-Sook
    • Archives of Pharmacal Research
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    • v.28 no.11
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    • pp.1244-1250
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    • 2005
  • The anti-inflammatory activity of Eurya emarginata (Thumb) Makino, of which leaves have been traditionally used to treat ulcers or diuretic in Jeju Island, has been investigated in the present study. Through the phytochemical study from the methanol extract of E. emaginiata, eutigosides Band C were isolated as the active components. Sseveral inflammatory markers including TNF-$\alpha$, IL-1$\beta$, IL-6, NO, iNOS, and COX-2 were examined. Eutigosides Band C potentially inhibited production of pro-inflammatory cytokines (IL-6 and TNF-$\alpha$) in a dose-dependent manner. Additionally, the intracellular contents of iNOS protein were markedly decreased after treatment with eutigosides Band C. The inhibition of iNOS activity was correlated with the decrease in nitrite levels. These results suggest that eutigoside Band C from E. emarginata may have anti-inflammatory activity through the inhibition of pro-inflammatory cytokines (TNF-$\alpha$ and IL-6), iNOS and COX-2.

The Effectiveness of Ulmus Davidiana Planch Herbal Acupuncture to Inhibit MIF Activation on Lipopolysaccharide-induced Rheumatoid Arthritis Model (유근피(楡根皮) 약침이 lipopolysaccharide 유발 류마티스 관절염 모델에서 MIF 활성 억제에 미치는 영향)

  • Byun, Hyuk;Park, In-Shik;Cho, Hyun-Seok;Kim, Kap-Sung;Lee, Seung-Deok
    • Journal of Acupuncture Research
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    • v.23 no.6
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    • pp.117-132
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    • 2006
  • Objectives : The purpose of this study is to investigate the effect of Ulmus davidiana Planch herbal acupuncture solution in LPS-stimulated RAW 264.7 cells and mouse knee joints, perfom1ed several experimental items: those are MIF mRNA, MIF, $TNF-{\alpha}$, $NF-{\kappa}B$ p65, iNOS mRNA, iNOS, NO, synovial hyperplasia, angiogenesis and fibrosis. Methods : In order to observe mRAN expression of MIF and iNOS in LPS-stimulated RAW 264.7 cells, RT-PCR was used. NO production in LPS-stimulated RAW 264.7 cells was measured by nitrite assay. All the female BALB/c mice were bred and maintained in pathogen-free mouse colonies and were 6 weeks of age on beginning of the experiment. The experimental model of RA was induced by injection of $50{\mu}g/kg$ LPS. Ulmus davidiana Planch herbal acupuncture solution was injected into either S 35 (犢鼻) or EX-LE 202 (內膝眼) of mice in turn daily for 19 days. Immunohistochemical staining was carried out to assess $TNF-{\alpha}$, $NF-{\kappa}B$ p65 and iNOS expression in synovial membrane. Synovial hyperplasia, angiogenesis and fibrosis in synovial membrane was observed with a microscope. Results : 1. Ulmus davidiana Planch herbal acupuncture solution inhibited mRNA expression of MIF and iNOS in dependence on a density of it in LPS-stimulated RAW 264.7 cells. 2. Ulmus davidiana Planch herbal acupuncture solution decreased synovial hyperplasia, angiogenesis and fibrosis in LPS-stimulated mouse knee joints. 3. Ulmus davidiana Planch herbal acupuncture solution curtailed production of MIF, $TNF-{\alpha}$, $NF-{\kappa}B$ p65, iNOS in LPS-stimulated mouse knee joints. Conclusion : On the basis of these results, It was shown that Ulmus davidiana Planch herbal acupuncture solution is significantly able to inhibit the production of MIF as a top in cytokines related to inflammatory or irrlll1une responses. Our results may provide that Ulmus davidiana Planch herbal acupuncture solution has beneficial effect in not only RA but other inflammatory or immune deases.

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Anti-inflammatory Effect of Nypa fruticans Wurmb. on tumor necrosis factor (TNF)-α-induced Inflammatory response in HaCaT cells (TNF-α로 유도된 HaCaT 각질형성세포의 염증반응에서 해죽순의 항염증 효과)

  • Bae, Gi-Sang;Park, Sung-Joo
    • The Korea Journal of Herbology
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    • v.34 no.1
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    • pp.51-57
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    • 2019
  • Objectives : Nypa fruticans Wurmb. (NF) have been used as a traditional medicine to treat inflammatory diseases in East-South Asia. However, it is largely undiscovered whether NF water extract could exhibit anti-inflammatory activities against tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$)-induced inflammatory responses on human keratinocytes, HaCaT cells. Therefore, this study was aimed to investigate the anti-inflammatory activity of NF water extract on TNF-${\alpha}$-induced inflammatory responses in HaCaT cells. Methods : To investigate the anti-inflammatory activites of NF water extract in HaCaT cells, the inflammatory model of HaCaT cells was established under a suitable concentration (10 ng/ml) of human TNF-${\alpha}$ (hTNF-${\alpha}$). HaCaT keratinocyte cells were pre-treated with NF water extract for 1 h, and then stimulated with hTNF-${\alpha}$. Then, the cells were harvested to measure the inflammatory mediators such as inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 and prostaglandin $E_2$ ($PGE_2$), and pro-inflammatory cytokine including TNF-${\alpha}$ and interleukin (IL)-6. In addition, we examined the inhibitory mechanisms of NF, mitogen activated protein kinases (MAPKs) and inhibitory kappa B alpha ($I{\kappa}-B{\alpha}$) Results : The treatment of NF inhibited the hTNF-${\alpha}$-induced elevation of iNOS, COX-2, and $PGE_2$ in HaCaT cells. In addition, NF treatment inhibited the hTNF-${\alpha}$-induced elevation of TNF-${\alpha}$ and IL-6. Furthermore, NF treatment inhibited the activation of MAPKs but not degradation of $I{\kappa}-B{\alpha}$. Conclusions : Taken together, our result suggest that treatment of NF could inhibit the hTNF-${\alpha}$-induced inflammatory responses via deactivation of MAPKs in HaCaT cells. This study could suggest that NF could be a beneficial agent to prevent skin damage or inflammation.

Inhibition of TNF-α-mediated NF-κB Transcriptional Activity in HepG2 Cells by Dammarane-type Saponins from Panax ginseng Leaves

  • Song, Seok-Bean;Tung, Nguyen Huu;Quang, Tran Hong;Ngan, Nguyen Thi Thanh;Kim, Kyoon-Eon;Kim, Young-Ho
    • Journal of Ginseng Research
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    • v.36 no.2
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    • pp.146-152
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    • 2012
  • Panax ginseng (PG) is a globally utilized medicinal herb. The medicinal effects of PG are primarily attributable to ginsenosides located in the root and leaf. The leaves of PG are known to be rich in various bioactive ginsenosides, and the therapeutic effects of ginseng extract and ginsenosides have been associated with immunomodulatory and anti-inflammatory activities. We examined the effect of PG leaf extract and the isolated ginsenosides, on nuclear factor (NF)-${\kappa}B$transcriptional activity and target gene expression by applying a luciferase assay and reverse transcription polymerase chain reaction in tumor necrosis factor (TNF)-${\alpha}$-treated hepatocarcinoma HepG2 cells. Air-dried PG leaf extract inhibited TNF-${\alpha}$-induced NF-${\kappa}B$transcription activity and NF-${\kappa}B$-dependent cyclooxygenase (COX)-2 and inducible nitric oxide synthase (iNOS) gene expression more efficiently than the steamed extract. Of the 10 ginsenosides isolated from PG leaves, Rd and Km most significantly inhibited activity in a dose-dependent manner, with $IC_{50}$ values of $12.05{\pm}0.82$ and $8.84{\pm}0.99\;{\mu}M$, respectively. Furthermore, the ginsenosides Rd and Km inhibited the TNF-${\alpha}$-induced expression levels of the COX-2 and iNOS gene in HepG2 cells. Air-dried leaf extracts and their chemical components, ginsenoside Rd and Km, are involved in the suppression of TNF-${\alpha}$-induced NF-${\kappa}B$ activation and NF-${\kappa}B$-dependent iNOS and COX-2 gene expression. Consequently, air-dried leaf extract from PG, and the purified ginsenosides, have therapeutic potential as anti-inflammatory.