• Title/Summary/Keyword: inhibition ELISA

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Glucosamine Inhibits Lipopolysaccharide-induced Inflammatory Responses in Human Periodontal Ligament Fibroblasts

  • Kim, Eun Dam;Park, Hyun-Jung;Baek, Jeong-Hwa
    • International Journal of Oral Biology
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    • v.39 no.4
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    • pp.221-228
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    • 2014
  • Glucosamine is commonly taken by the elderly without prescription as a nutritional supplement to attenuate the progression or symptoms of osteoarthritis. Previous studies demonstrated that glucosamine shows anti-inflammatory effects in tissues such as blood vessels and the heart. However, there have been few reports about the effects of glucosamine on oral inflammatory diseases. Therefore, in this study, the effects of glucosamine on lipopolysaccharide (LPS)-induced inflammatory responses were investigated using human periodontal ligament fibroblasts (HPDLFs). HPDLFs were incubated in the presence and absence of glucosamine (10 mM) for 24 h, followed by treatment with E. coli LPS (100 ng/ml) or vehicle. Quantitative RT-PCR and ELISA results showed that LPS exposure significantly increased the levels of IL-6 and IL-8 mRNA and protein, while the effect was significantly suppressed by glucosamine treatment. Glucosamine did not attenuate, but slightly increased, the LPS-induced activation of mitogen activated kinases (ERK, p38, JNK). However, it suppressed the LPS-induced increase in the DNA binding affinity and transcriptional activity of NF-${\kappa}B$. These results suggest that glucosamine exerts anti-inflammatory effects on HPDLFs exposed to LPS via inhibition of NF-${\kappa}B$ activity, necessitating further studies using animal periodontitis models.

Anti-inflammatory Effect of Mugi-hwan Water Extract in RAW 264.7 Cells (무기환(戊己丸)의 RAW 264.7 세포에 대한 항염증작용 연구)

  • Kim, Ilhyun;Choi, Chonghwan;Lee, Sewon;Song, Yungsun
    • Journal of Korean Medicine Rehabilitation
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    • v.23 no.3
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    • pp.27-35
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    • 2013
  • Objectives The aim of this study was to investigate anti-inflammatory activity of Mugi-hwan (MGH) Water Extract. Methods Cells were treated with 2 ug/ml of LPS 1 hour prior to the addition of MGH. Cell viability was measured by MTS assay. The production of NO was determined by reacting cultured medium with Griess reagent. The expression of COX-2, iNOS and MAPKs was investigated by Western blot, RT-PCR. The content of level of cytokines ($PGE_2$, IL-6, in media from LPS-stimulated Raw 264.7 cells was analyed by ELISA kit. Results MGH inhibited the production of NO, $PGE_2$, IL-6 as well as the expressions of iNOS, COX-2 in the murine macrophage, RAW 264.7 cells. MGH also had suppression effects of LPS induced MAPKs activation. Conclusions These results suggest that MGH has an anti-inflammatory therapeutic potential, which may result from inhibition of MAPK phosphorylation, thereby decreasing the expression of pro-inflammatory genes.

Vp28 of Shrimp White Spot Syndrome Virus Is Involved in the Attachment and Penetration into Shrimp Cells

  • Yi, Guohua;Wang, Zhimin;Qi, Yipeng;Yao, Lunguang;Qian, Juan;Hu, Longbo
    • BMB Reports
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    • v.37 no.6
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    • pp.726-734
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    • 2004
  • White spot disease (WSD) is caused by the white spot syndrome virus (WSSV), which results in devastating losses to the shrimp farming industry around the world. However, the mechanism of virus entry and spread into the shrimp cells is unknown. A binding assay in vitro demonstrated VP28-EGFP (envelope protein VP28 fused with enhanced green fluorescence protein) binding to shrimp cells. This provides direct evidence that VP28-EGFP can bind to shrimp cells at pH 6.0 within 0.5 h. However, the protein was observed to enter the cytoplasm 3 h post-adsorption. Meanwhile, the plaque inhibition test showed that the polyclonal antibody against VP28 (a major envelope protein of WSSV) could neutralize the WSSV and block an infection with the virus. The result of competition ELISA further confirmed that the envelope protein VP28 could compete with WSSV to bind to shrimp cells. Overall, VP28 of the WSSV can bind to shrimp cells as an attachment protein, and can help the virus enter the cytoplasm.

Inhibitory Effect of Lignans from the Rhizomes of Coptis japonica var. dissecta on Tumor Necrosis Factor-${\alpha}$ Production in Lipopolysaccharide-stimulated RAW264.7 Cells

  • Cho, Jae-Youl;Park, Ji-Soo;Yoo, Eun-Sook;Kazuko Yoshikawa;Baik, Kyong-Up;Lee, Jong-Soo;Park, Myung-Hwan
    • Archives of Pharmacal Research
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    • v.21 no.1
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    • pp.12-16
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    • 1998
  • The inhibitory effect of 10 lignan constituents isolated from the rhizomes of Coptis japonica var. dissects on tumor necrosis factor (TNF)-${\alpha}$ production in lipopolysaccharide (LPS)-stimulated macrophage cell line (RAW264.7 cells) has been studied. Among them, pinoresinol, woorenoside-V and lariciresinol glycoside showed significant inhibitory activities in the range from 37% to 55% at the concentration of $25{\mu}g/ml.$ The results are first report that the lignans isolated from Coptis japonica inhibit TNF- ${\alpha}$${\alpha}$

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Anti-adipogenic Effect of Chlorogenic Acid in 3T3-L1 Adipocytes

  • Park, Se-Eun;Choi, Jun-Hui;Lee, Hyo-Jeong;Seo, Kyoungsun;Kim, Seung
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.80-80
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    • 2018
  • Chlorogenic acid is a phenolic compound found in Cudrania tricuspidata fruits. In the present study, the effect of chlorogenic acid on the inhibition of adipogenesis in 3T3-L1 adipocytes was investigated. Cells were stained with Oil red O reagent to detect lipid droplets in adipocytes. The 3T3-L1 cells were lysed and measured for intracellular triglyceride and adipokine by ELISA kit. The protein expression of adipogenesis-related gene was evaluated by Western blot analysis. Chlorogenic suppressed lipid droplet and intracellular triglyceride accumulation in a concentration manner and also decreased secretion of adipokines such as leptin and adiponectin, compared with fully differentiated adipocytes. Treatment of 3T3-L1 cells with chlorogenic acid reduced the protein levels of peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) and, CCAAT/enhancer binding proteins alpha ($C/EBP{\alpha}$). This indicates that chlrogenic acid was effective as an anti-obesity agent by repressing the differentiation of 3T3-L1 into adipocytes and inhibiting triglyceridef formation in adipocyte and that it exerts its role mainly through the significant down-regulation of $PPAR{\gamma}$ and $C/EBP{\alpha}$.

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Anti-inflammatory Effects of Abeliophyllum distichurn Flower Extract

  • Lee, Jin Wook;Kang, Yoon Joong
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.89-89
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    • 2018
  • Abeliophyllum distichum is a medicinal plant used in regional traditional medicine to relieve pain in inflammatory processes. In this study, anti-inflammatory effects of Abeliophyllum distichum stem (ADS) ethyl acetate extract were examined. Furthermore, possible molecular mechanisms of the anti-inflammatory effects were dissected. The anti-inflammatory activity was investigated by inhibition of lipopolysaccharide (LPS) induced pro-inflammatory cytokine production in murine macrophage-like cell line Raw264.7 cells and human microglial cell line BV2 cells. The measurement of the induced pro-inflammatory cytokine levels were carried out by ELISA. The phosphorylation of ERK1/2, JNK, and MAPK, and the nuclear expression of nuclear factor $NF-{\kappa}B$ p65 were investigated by Western blot analysis. The extract of ADS significantly decreased the production of pro-inflammatory cytokines. In addition, the extract suppressed the phosphorylation of ERK1/2, JNK, and p38 MAPK, and the nuclear translocation of $NF-{\kappa}B$ p65 in activated cells. Our findings provide evidence for the popular use of Abeliophylli distichum in inflammation around Goesan region and also suggest that the stem extract has potential therapeutic benefits against several inflammatory diseases.

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Anti-inflammatory Effects of Kochiae Fructus Extract on LPS-stimulated Raw 264.7 Cells

  • Kim, Bit-Na;Seong, Eun-Hwa;Kang, Yoon Joong
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2018.04a
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    • pp.90-90
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    • 2018
  • Kochiae Fructus is a medicinal plant used as medicine of the urinary organs, skin disease and inflammation. In this study, anti-inflammatory activity of Kochiae Fructus extract and its possible mechanisms of action were examined. The anti-inflammatory activity was investigated by inhibition of lipopolysaccharide (LPS) induced nitric oxide (NO), pro-inflammatory cytokine production and matrix metalloprotease-9 (MMP-9) in murine macrophage-like cell line Raw 264.7 cells. The measurement of the induced pro-inflammatory cytokine levels were carried out by ELISA. The phosphorylation of ERK1/2, JNK and MAPK and the nuclear expression of nuclear factor $NF-{\kappa}B$ p65 were investigated by Wesern blot analysis. The extract suppressed the phosphorylation of ERK1/2, JNK, and p38 MAPK, and the nuclear translocation of $NF-{\kappa}B$ p65 in activated cells. As a result, we suggest that the extract of Kochiae Fructus decreased the production of pro-inflammatory cytokines ($TNF-{\alpha}$, IL-6), nitric oxide, MMP-9 in LPS-induced Raw264.7 cells. Kochiae Fructus has possibility to be used as therapeutic benefits against inflammatory diseases.

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Anti-inflammatory activity of methanol extract isolated from stem bark of Albizia julibrissin

  • Na, Ho-Jeong;Cha, Dong-Seok;Jeon, Hoon
    • Advances in Traditional Medicine
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    • v.9 no.2
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    • pp.157-163
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    • 2009
  • Albizia julibrissin (AJ) has been used widely as a traditional medicine. In macrophages nitric oxide (NO) is released as an inflammatory mediator and has been proposed to be an important modulator of many pathophysiological conditions including inflammation and carcinogenesis. In this study we have examined the NO inhibition effect of 85% methanol extracts of AJ in mouse macrophage. Lipopolysaccharide (LPS) has been reported to induce production of NO. Extracts of AJ (1, 10, $100{\mu}g/ml$) suppressed nitric oxide production in LPS-stimulated ($100{\mu}g/ml$) mouse (C57BL/6) macrophages and analyzed by ELISA. In addition, it also attenuated the expression of inflammatory products like Interleukin-6 (IL-6), cyclooxygenase-2 (COX-2), and inducible NOS (iNOS) as assessed by immunoblotting with specific antibodies. These results suggest that 85% methanol extracts of AJ would be useful in inflammatory diseases.

The Effect of Silibinin Extracted from Cirsium Japonicum on Allergic Inflammation (대계(大薊)의 주성분인 Silibinin이 알레르기 염증반응에 미치는 효과(效果))

  • Kim, Beom-Rak;Kim, Koung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.23 no.1
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    • pp.44-58
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    • 2010
  • Silibinin is the major active molecule of silymarin, the mixture of flavonolignans extracted from Cirsium japonicum (CJ). It has been used for treatment of hepatitis and inflammation related diseases. The aim of this study was to prove whether Silibinin has effectiveness for allergic inflammation. Silibinin processes the inflammatory reaction in phorbol 12-myristate 13-acetate (PMA) plus calcium ionophore A23187 (PMA plus A23187) stimulated human mast cell line (HMC-1). Its effect was examined by ELISA, RT-PCR, Western blot, and Luciferase assay. The results were Silibinin inhibited the expression of histamine, TNF-$\alpha$ (tumor necrosis factor-$\alpha$), IL-6 (interleukin-6), and IL-8 (interleukin-8). Silibinin suppressed NF-${\kappa}B$ (nuclear factor kappa B) activation in stimulated HMC-1 (human mast cell-1). This effect was mediated through inhibition of phosphorylation and degradation of $IkB{\alpha}$, an inhibitor of NF-kB. Silibinin significantly inhibited induction of NF-kB promoter mediated Luciferase assay. These results suggest that Silibinin has a potential molecule for therapy of mast cell-derived allergic inflammatory diseases.

Inhibitory Effect of Rhododendron Mucronulatum Root Extract on Allergic Inflammation (진달래 뿌리 추출물의 알레르기 염증 억제 효과)

  • Jang, Si Sung;Lee, DaeJoong;Song, Jihoon;Park, Do Hwi;Jeon, Chan Yong;Hwang, Gwi Seo
    • The Journal of Internal Korean Medicine
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    • v.43 no.1
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    • pp.68-78
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    • 2022
  • Objective: In this study, we investigated the protective effect of rhododendron mucronulatum extract (RME) on allergic reactions and inflammation. Methods: The effect of RME was determined using ELISA and RT-PCR in RBL-2H3 mast cells and RAW 264.7 macrophage cells. We determined cell viability, β-hexosaminidase release, and the synthesis of IL-4 and TNF-α in RBL-2H3 cells. In addition, we determined NO from RAW 264.7 and the gene expression of IL-1β, iNOS, IL-6, TNF-α, and IL-10. Results: RME inhibited β-hexosaminidase release and synthesis of IL-4 and TNF-α in RBL-2H3 by the anti-DNP IgE plus DNP-HSA stimulation. In addition, RME inhibited the production of NO and the gene expression of IL-1β, iNOS, IL-6, and TNF-α in LPS-stimulated RAW 264.7 cells. Conclusion: From these results, we concluded that RME possesses anti-allergic activity and anti-inflammatory activity due to the inhibition of mast cells and macrophage function.