• Title/Summary/Keyword: RAW 264.7 murine macrophage

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Stimulation of the Immune Response by Yanggyuksanhwa-tang (소양인 양격산화탕의 면역 활성화 연구)

  • Jung, Da-Young;Ha, Hye-Kyung;Lee, Ho-Young;Lee, Jin-Ah;Lee, Nam-Hun;Lee, Jun-Kyoung;Huang, Dae-Sun;Shin, Hyeun-Kyoo
    • Journal of Sasang Constitutional Medicine
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    • v.22 no.4
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    • pp.77-84
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    • 2010
  • 1. Objectives Yanggyuksanhwa-tang for Soyangin was applied to investigate the immunological activities on antigen (Ag)-specific or Ag-non-specific immune responses on murine macrophage cell line (RAW 264.7) and ovalbumin/aluminium (OVA/Alum)-immunized mice. 2. Methods This study were carried out in nitric oxide (NO) synthesis on RAW 264.7 cells and cellular proliferation on mouse splenocytes. C57BL/6 mice were immunized intraperitonially with OVA/Alum (100 ${\mu}g$/200 ${\mu}g$) on day 1, 8, and 15. Yanggyuksanhwa-tang was administrated to mice orally for 3 weeks from day 1. On day 22, OVA-, lipopolysaccharide (LPS)-, and concanavalin A (Con A)-stimulated splenocyte proliferation and antibodies (OVA-specific antibodies of the IgG, IgG1, and total IgM classes) in plasma were measured. 3. Results Yanggyuksanhwa-tang significantly enhanced cellular proliferation by LPS and Con A on splenocytes from OVA/Alum-immunized mice (p<.001). Yanggyuksanhwa-tang also significantly enhanced plasma OVA-specific IgG (p<.001), IgG1 (p<.001), and total IgM (p<.01) levels compared with the OVA/Alum group. 4. Conclusions These results suggested that Yanggyuksanhwa-tang for Soyangin could be used as immunopotent.

Codium fragile Ethanol Extraction Inhibited Inflammatory Response through the Inhibition of JNK Phosphorylation

  • Han, Sin-Hee;Kim, Young-Guk;Lee, Su-Hwan;Park, Chung-Berm;Choi, Han-Gil;Jang, Hye-Jin;Lee, Young-Seob;Kwon, Dong-Yeul
    • Preventive Nutrition and Food Science
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    • v.15 no.3
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    • pp.206-212
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    • 2010
  • Codium fragile (CF) is an edible green alga consumed as a traditional food source in Korea. In this study, the ethanol extract of CF was evaluated to determine if it has anti-inflammatory activity. Lipopolysaccharide (LPS), a toxin from bacteria, is a potent inducer of inflammatory cytokines, such as tumor necrosis factor (TNF)-$\alpha$ and interleukin (IL)-6. Therefore, we studied whether CF extracts have an anti-inflammatory effect in LPS-induced murine macrophage cell lines (RAW 264.7). In the present study, IL-6 production was measured using an enzyme-linked immunosorbent assay (ELISA), prostaglandin $E_2$($PGE_2$) production was measured using the EIA kit, and cyclooxygenase (COX)-2 and mitogen-activated protein kinase (MAPK) activation were determined by Western blot analysis. IL-6 mRNA, COX-2 mRNA and iNOS mRNA expression were measured using reverse transcription-polymerase chain reaction (RT-PCR). The results indicated that CF extracts inhibit LPS-induced IL-6, NO and PGE2 production in a dose-dependent manner, as well as expression of iNOS and COX-2. CF extracts significantly inhibited LPS-induced c-Jun N-terminal kinase (JNK) 1/2 phosphorylation. Taken together, these findings may help elucidate the mechanism by which CF modulates RAW 264.7 cell activation under inflammatory conditions.

Anti-inflammatory activity of Ganoderma lucidum by inhibition of NF-κB p65 phosphorylation

  • Kim, Hyung Don;Park, Jeong-Yong;Noh, Hyung-Jun;Lee, Seung Eun;Lee, Jeong Hoon;Seo, Kyung Hye
    • Korean Journal of Agricultural Science
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    • v.46 no.3
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    • pp.653-660
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    • 2019
  • Ganoderma lucidum, an oriental polypore fungus and medicinal mushroom, has a long history of use for promoting health and longevity in Korea, China, and other Asian countries. This study was aimed at determining the anti-inflammatory activity and mechanism of action of Ganoderma lucidum in murine macrophage RAW 264.7 cells. Ganoderma lucidum was extracted with ethanol and freeze-dried. The anti-inflammatory effect (nitrite production) of Ganoderma lucidum extracts was tested using a nitric oxide (NO) colorimetric assay. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) was performed to quantify the mRNA expression of cytokines including tumor necrosis factor-${\alpha}$ ($TNF-{\alpha}$), interleukin $(IL)-1{\beta}$, and IL-6. Western blotting was performed to measure the expression levels of inflammation-related proteins, such as inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), nuclear factor kappa B ($NF-{\kappa}B$) p65, and phosphorylated $NF-{\kappa}B$ p65. The NO colorimetric assay showed that NO production increased with the treatment of lipopolysaccharide in (LPS)-activated RAW 264.7 macrophages and decreased with the cotreatment of Ganoderma lucidum extracts and LPS. Ganoderma lucidum extracts repressed the mRNA expressions of cytokines, which were increased after the LPS treatment. In addition, Ganoderma lucidum extracts inhibited the LPS-induced expression of iNOS and COX-2 and the LPS-induced phosphorylation of $NF-{\kappa}B$ p65. These results suggest that the Ganoderma lucidum extracts exert an anti-inflammatory activity by inhibiting $NF-{\kappa}B$ related proteins and cytokines.

Investigation of Pharmacological Stability of Hyeonggaeyeongyotang Gagambang by Formulation Changes for Foaming Tablet (형개연교탕(荊芥連翹湯) 가감방(가감방)의 발포정 제형 변화에 따른 약리학적 안정성 연구)

  • Cho, Nam Joon;Gang, Seong Gu;Kim, Ji Young;Han, Hyo Sang;Kim, Kee Kwang
    • The Korea Journal of Herbology
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    • v.33 no.5
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    • pp.67-72
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    • 2018
  • Objectives : Natural extracts have been extensively studied to replace single agent drugs that cause a variety of side effects. However, studies of changes to the formulation of natural extracts has not been nearly proceed. We aimed to investigate whether pharmacological stability of hyeonggaeyeongyotang gagambang (HYT) is altered by formulation changes for foaming tablet. Methods : In this study, we performed freeze - drying of HYT, which is known to have antioxidant and anti - inflammatory properties, and then changed the formulation by foaming. Results : As a result, the foaming reaction appeared normally when HYT foamed tablets were put into water, and almost all of the substances were dissolved in the aqueous solution. In addition, we confirmed using high-performance liquid chromatograph that the geniposide used as an indicator material of HYT was stable in most of the formulations. It was confirmed that the change of HYT formulation did not affect the antioxidant efficacy by the 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid assay. Moreover, quantitative real-time PCR confirmed that the inhibitory effect of HYT on IL-$1{\beta}$ mRNA expression induced by lipopolysaccharides treatment in murine macrophage RAW 264.7 cells was similar in the solution of foaming tablet. Conclusions : These results suggest that the materials with various pharmacological effects can be stably maintained even when the formulation is changed by the foaming action of HYT. Our results are expected to provide important basic knowledge on formulation changes using various natural extracts.

Antioxidant and Anti-inflammatory Effects of Ficus erecta var. sieboldii Leaf Extract in Murine Macrophage RAW 264.7 Cells (좁은잎천선과나무(Ficus erecta var. sieboldii) 잎 추출물이 대식세포 RAW 264.7 세포에서 미치는 항산화 및 항염증 효과)

  • Jung, Yong-Hwan;Ham, Young-Min;Yoon, Seon-A;Oh, Dae-Ju;Kim, Chang-Suk;Yoon, Weon-Jong
    • Korean Journal of Plant Resources
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    • v.31 no.4
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    • pp.303-311
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    • 2018
  • In this study, a preliminary evaluation of the antioxidant and anti-inflammatory activity of the Ficus erecta var. sieboldii (Miq.) King (FES) leaf extract has been performed to assess its potential as a natural resource for food and medicinal materials. FES was extracted using 70% EtOH and then fractionated sequentially using n-hexane, $CH_2Cl_2$, EtOAc, and n-BuOH. To screen for antioxidant and anti-inflammatory agents effectively, the inhibitory effect of the FES extracts on the production of oxidant stresses (DPPH, xanthine oxidase, and superoxide) and pro-inflammatory factors (NO, iNOS, COX-2, $PGE_2$, IL-6, and $IL-1{\beta}$) in the murine macrophage cell line RAW 264.7 activated with lipopolysaccharide (LPS) was examined. Among the sequential solvent fractions of FES, the $CH_2Cl_2$ and EtOAc fractions showed decreased production of oxidant stresses (DPPH, xanthine oxidase and superoxide), and the hexane and $CH_2Cl_2$ fractions of FES inhibited the production of pro-inflammatory factors (NO, iNOS, COX-2, and $PGE_2$). The $CH_2Cl_2$ fraction also inhibited the production of pro-inflammatory cytokines ($TNF-{\alpha}$, IL-6, and $IL-1{\beta}$). These results suggest that FES has a significant effects on the production of oxidant stresses and pro-inflammatory factors and may be used a natural resource for antioxidant and anti-inflammatory agents.

A New Ceanothane-Type Triterpenoid Saponin Isolated from Gouania leptostachya DC. var. tonkinensis Pit. and Its Underlying Anti-Inflammatory Effects

  • Nguyen Thi Hang;Nguyen Thi Bich Thu;Le Ba Vinh;Nguyen Viet Phong;Tran Van On;Ki Yong Lee
    • Journal of Microbiology and Biotechnology
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    • v.33 no.7
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    • pp.941-948
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    • 2023
  • Metabolites from medicinal plants continue to hold significant value in the exploration and advancement of novel pharmaceuticals. In the search for plants containing compounds with anti-inflammatory effects, we observed that the ethanol (EtOH) extract obtained from the aerial components of Gouania leptostachya DC. var. tonkinensis Pit. exhibited substantial suppression of nitric oxide (NO) in vitro. In a phytochemical study on an EtOH extract of G. leptostachya, 11 compounds were purified, including one unreported compound namely gouanioside A (1). Their chemical structures were unambiguously determined through the use of various spectroscopic techniques, such as 1 and 2D NMR, IR, and HR-ESI-MS, and by producing derivatives via chemical reactions. The EtOH extract, fractions, and a new compound exerted inflammatory effects by altering NO synthesis in murine RAW264.7 macrophage cells stimulated with lipopolysaccharide. The underlying inflammatory mechanism of the new compound 1 was also explored through various in vitro experiments. The results of this study indicate the potential usefulness of new compound 1 from G. leptostachya as a treatment for inflammatory diseases.

Phenolic Compound Content and Antioxidant Activity of Citrus Peels (감귤 과피의 페놀성 화합물 함량과 항산화 활성)

  • Hwang, Joon-Ho;Park, Kyeong-Yeol;Oh, You-Sung;Lim, Sang-Bin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.2
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    • pp.153-160
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    • 2013
  • The peel from seven types of citrus was extracted with 80% methanol, and their phenolic compound content, oxygen radical absorbance capacity (ORAC), inhibitory activities of nitric oxide (NO), and reactive oxygen species (ROS) production induced by LPS and t-BHP in LPS-activated RAW 264.7 cells were measured. Total phenolic content was high in Yungkyool, Cheonhyehyang, and Jinkyool (30.6, 30.2, and 28.2 mg GAE/g, respectively), while total flavonoid content was high in Yungkyool and Jinkyool (30.3 and 25.5 mg RE/g, respectively). ORAC was the highest at 1,076 mM TE/g in Yungkyool, followed by Cheonhyehyang (1,012), Jinkyool (984), and Hallabong (914). High inhibitory activity against NO production was shown in Cheonhyehyang, Yungkyool, and Jinkyool with $IC_{50}$ values of 215.3, 259.2, and 328.9 ${\mu}g/mL$, respectively. LPS-induced ROS production was inhibited by 16.4% and 12.8% in Hallabong and Jinkyool, while t-BHP-induced ROS production was inhibited by 28.7%, 26.1%, and 26.6% in Jinkyool, Hallabong, and Cheonhyehyang, respectively. Correlation coefficients between total phenolic, total flavonoid content, and ORAC were 0.884 and 0.855. Inhibitory activity against NO production showed higher correlation with total phenolic content than total flavonoid content. It was concluded that citrus peels had potent antioxidant activities and could be used as natural antioxidants in the food and pharmaceutical industries.

Immunomodulatory Activity of Water Extract of Ulmus macrocarpa in Macrophages (유근피 추출물이 대식세포 면역조절에 미치는 영향)

  • Kwon, Da Hye;Kang, Hye-Joo;Choi, Yung Hyun;Chung, Kyung Tae;Lee, Jong Hwan;Kang, Kyung Hwa;Hyun, Sook Kyung;Kim, Byung Woo;Hwang, Hye Jin
    • Journal of Life Science
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    • v.26 no.1
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    • pp.50-58
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    • 2016
  • The root bark of Ulmus macrocarpa has been used in traditional medicine for the treatment of various diseases such as edema, infection and inflammation. Nevertheless, the biological activities and underlying mechanisms of the immunomodulatory effects remain unclear. In this study, as part of our ongoing screening program to evaluate the immunomodulatory potential of new compounds from traditional medicinal resources, we investigated the effects of U. macrocarpa water extract (UME) on immune modulation in a murine RAW 264.7 macrophage model. As immune response parameters, the productions of as nitric oxide (NO) and cytokines such tumor necrotic factor (TNF)-α, interleukin (IL)-1β and IL-10 were evaluated. Although the release of IL-1β remained unchanged in UME-treated RAW 264.7 macrophages, the productions of NO, TNF-α and IL-10 were significantly increased, along with the increased expression of inducible NO synthase, TNF-α and IL-10 expression at concentrations with no cytotoxicity. UME treatment also induced the nuclear translocation of nuclear factor κB (NF-κB), and phosphorylation of Akt and mitogen-activated protein kinases (MAPKs) indicating that UME activated macrophages through the activation of NF-κB, phosphoinositide-3-kinase (PI3K)/Akt and MAPKs signaling pathways in RAW 264.7 macrophages. Furthermore, pre-treatment with UME significantly attenuated the production of NO, but not TNF-α, IL-1β and IL-10, in lipopolysaccharide-stimulated RAW 264.7 cells suggesting that UME may be useful in preventing inflammatory diseases mediated by excessive production of NO. These findings suggest that the beneficial therapeutic effects of UME may be attributed partly to its ability to modulate immune functions in macrophages.

Anti-inflammatory Effects of Purpurogallin Carboxylic Acid, An Oxidation Product of Gallic Acid in Fermented Tea (발효차중의 미량 성분인 gallic acid 산화물 purpurogallin carboxylic acid의 항염증 효과)

  • Jhoo, Jin-Woo
    • Korean Journal of Food Science and Technology
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    • v.40 no.6
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    • pp.707-711
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    • 2008
  • The principal objective of the current study was to isolate a purpurogallin derivative as an oxidation product from gallic acid, in an effort to assess the anti-inflammatory effects of this compound. Purpurogallin derivative is known to be the one of the oxidation products of gallic acid. This compound has been identified as a minor chemical component in fermented tea products. It has been previously demonstrated that theaflavins, the oxidation products of catechins found in fermented tea products, exert profound antioxidant and anti-inflammatory effects. However, the biological activities of a minor chemical component in fermented teas have yet to be evaluated. Purpurogallin carboxylic acid (PCA) was identified as a major oxidation product of gallic acid from a peroxidase/hydrogen peroxide oxidation model system. The identity of the PCA was verified by $^{1}H$ NMR, $^{13}C$ NMR and MS techniques. PCA treatment significantly suppressed the generation of pro-inflammatory mediators including nitric oxide and IL-6 in lipopolysaccharide (LPS)-stimulated RAW264.7 murine macrophages. According to the nitrite assay, PCA 100, 75, and $50{\mu}g/mL$ treatment dose-dependently inhibited NO production by 57.6, 41.5, and 21.8%, respectively, in LPS-stimulated RAW264.7 murine macrophage cells. Moreover, IL-6 production was inhibited to a significant degree with PCA treatment of 100 and $75{\mu}g/mL$ at 43.1 and 23.9%, respectively. PCA treatment also significantly suppressed $PGE_2$ production at levels of 100 and $75{\mu}g/mL$. These results showed that PCA exerts inhibitory effects on the production of inflammatory mediators.

Effects of Substance P on the Activities of Immune Cell (면역세포 활성에 대한 Substance P의 영향)

  • Kim, Hyung-Seop;Oh, Kwi-Ok;Lim, Chong-Deuk
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.376-395
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    • 1996
  • The neuropeptide substance P(SP) has been recognized to modulate immune systems, with close proximity between peptidergic sensory nerve endings and immune cells. These include the macrophage and neutrophil activation, IL-2 production in T cell, augmentation of Ig synthesis, mast cell degranulation, $PGE_2$ and collagenase secretion in synoviocytes. In this study I examined SP-induced various biological activities such as antimicrobial action, cytokine production, and mast cell degranulation in the presence or absence of other inflammatory cell activators. Antimicrobial studies showed that undifferentiated HL-60 cells were not affected by SP. However, SP significantly enhanced antimicrobial action of TPA-treated or dbcAMP-treated HL-60 cells which had been differentiated into PMN or macrophage/monocyte. I could not find synergistic relationship between SP and LPS in parallel experiments of the above. SP did not induce IL-l production from murine macrophage cell line RAW264.7 whether costimulated with LPS or not. Mast cell degranulation was occured only when stimulated with high dose ($10^{-5}M$) of SP and the degree of this activation was slightly reduced by simultaneous application of $MIP-1{\alpha}$. In addition, CGRP which is known to be a common coexisting neuropeptide with SP within specific fibers did not augment the function of SP on mast cell degranulation. These results suggest that immunoregulatory activities of SP could be mediated through direct upregulation of various functions of immune cells and also upregulation of responsiveness of immune cells to other immune activators.

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