• Title/Summary/Keyword: Macrophage cells

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Anti-inflammatory effects of seed ethanolic extracts of the common buckwheat and tartary buckwheat are mediated through the suppression of inducible nitric oxide synthase and pro-inflammatory cytokines in LPS-induced RAW 264.7 macrophage cells (일반메밀과 쓴메밀 종실 추출물의 RAW 264.7 대식세포에서 LPS에 의해 유도되는 iNOS 및 염증성 사이토카인 발현 저해를 통한 항염증 효과 비교)

  • Kim, Su Jeong;Sohn, Hwang Bae;Lee, Kyung-Tae;Shin, Ji-Sun;Kim, Suyeon;Nam, Jung Hwan;Hong, Su Young;Suh, Jong Taek;Chang, Dong Chil;Kim, Yul Ho
    • Korean Journal of Food Science and Technology
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    • v.51 no.6
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    • pp.565-575
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    • 2019
  • The ethanolic seed extracts of the common buckwheat (CB) and tartary buckwheat (TB) were examined for their anti-oxidant and anti-inflammatory effects on lipopolysaccharide (LPS)-induced RAW 264.7 cells. In this study, it was observed that the rutin content of TB extracts was 65-78 times higher than the CB extracts, while quercetin was only detected in the TB extracts. In addition, TB extracts were observed to have 1.8-2.0 times higher flavonoid and polyphenolic content than the CB extracts. Cytotoxicity was not observed when both the buckwheat extracts were evaluated at concentrations in the range of 6.25-400 ㎍/mL. The treatment with TB extracts significantly suppressed the LPS-induced nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) expression at the protein and mRNA levels. The TB extracts more potently inhibited the LPS-induced production of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, and IL-6 than the CB extracts. The mRNA levels of TNF-α, IL-1β, and IL-6 were also significantly inhibited both by the TB and CB extracts in a pattern similar to their production.

Anti-inflammatory Activities Verification of Ambrosia trifida L. extract in RAW 264.7 Cells (RAW 264.7 세포에서의 단풍잎돼지풀 추출물의 항염증 활성 검증)

  • Yoo, Dan-Hee;Lee, Jin-Young
    • Microbiology and Biotechnology Letters
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    • v.48 no.1
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    • pp.79-89
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    • 2020
  • This study was performed to evaluate the anti-inflammatory activities of 70% ethanol extract from Ambrosia trifida L. (AT). The electron donating ability and ABTS+ radical scavenging ability of extract from AT was shown to be 84.1% and 92.5% at 1,000 ㎍/ml concentration. The astringent effect of extract from AT was shown to be 94.7% at 1,000 ㎍/ml. The anti- inflammatory activities of extract of AT were investigated using RAW 264.7 cells induced by lipopolysaccharide (LPS). The cell toxicity effect of AT extract on RAW 264.7 performed MTT assay. As a result of the measured cell toxicity effect, 90% or more was shown with cell viability at a 500 ㎍/ml concentration. In nitric oxide synthesis inhibition effect, it was shown that extract from AT concentration dependent inhibited nitric oxide production. The protein expression inhibitory effect of AT extract was measured by western blot at 25, 50, and 100 ㎍/ml concentration and the β-actin used as a positive control. Consequently, the inducible nitric oxide synthase (iNOS), cyclooxygenase (COX)-2 protein expression inhibitory effect was decreased by 8.6%, 25.1% at 100 ㎍/ml concentration. The phosphorylation of extracellular signal-regulated kinase 1/2, p38, c-Jun NH2-terminal kinase and Iκ-Bα protein expression inhibitory effect was a decreased dependent concentration. The mRNA expression inhibitory effect was measured by reverse transcription - polymerase chain reaction at 25, 50, and 100 ㎍/ml concentration and the glyceraldehyde-3-phosphate dehydrogenase used as a positive control. Consequently, the iNOS, COX-2, interleukin (IL)-1β, IL-6 and tumor necrosis factor-α mRNA expression inhibition effect was a decreased dependent concentration in an LPS-activated macrophage. In conclusion, AT extract may have some effects on inflammatory factors as potential anti-inflammatory agents and natural substance for cosmetics.

Structural Characteristics and Anti-inflammatory Activities of Chemically Sulfated-hyaluronic Acid from Streptococcus dysgalactiae (Streptococcus dysgalactiae로부터 분리된 히알루론산과 황화된 유도체의 구조와 항염증 활성)

  • Hong, Chang-Il;Jung, Eui-Gil;Han, Kook-Il;Kim, Yong Hyun;Lee, Sung Hee;Lee, Hong Sub;Han, Man-Deuk
    • Journal of Life Science
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    • v.26 no.5
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    • pp.545-554
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    • 2016
  • Hyaluronic acid (HA) is an important macromolecule in medical and pharmaceutical fields. HA is a natural and linear polymer composed of repeating disaccharide units of β-1, 3-N-acetyl glucosamine and β-1, 4-glucuronic acid. This work aimed to confirm the structural characteristics and anti-inflammatory activities of HA and its chemically sulfated-HA. HA was produced from a fed-batch fermentation process using Streptococcus dysgalactiae in a 5 l bioreactor. HA was isolated water-soluble form (HA-WS) and water-insoluble form (HA-WI) from culture medium, and was obtained chemically sulfated-derivative (S-HA) that resulted in a 90% yield from HA-WI. The structural features of the sulfated- HA (S-HA) were investigated by FT-IR and 1H-NMR spectroscopy. The FT-IR and NMR patterns revealed the similarity in both the FTIR spectrum as well as NMR spectrum of both reference standard and purified HA from S. dysgalactiae. The anti-inflammatory activities of HA and S-HA were examined on LPS-induced RAW 264.7 cells. S-HA was significantly inhibited production of pro-inflammatory mediators such as nitric oxide (NO) and PGE2 and the gene levels of iNOS and COX-2, which are responsible for the production of NO and PGE2, respectively. Furthermore, S-HA also suppressed the overproduction of pro-inflammatory cytokine TNF-α (<80 pg/ml) and IL-6 (<100 pg/ml) compared to that of HA-WI. The present study clearly demonstrates that HA-S exhibits anti-inflammatory activities in RAW 264.7 macrophage cells.

Quality Characteristics and Biological Activity of Fermented Black Garlic with Probiotics (Probiotics를 이용한 흑마늘 발효물의 품질특성 및 생리활성)

  • Tak, Hyun-Min;Kim, Gyeong-Min;Kim, Jong-Su;Hwang, Cho-Rong;Kang, Min-Jung;Shin, Jung-Hye
    • Journal of Life Science
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    • v.24 no.5
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    • pp.549-557
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    • 2014
  • This study examined the quality characteristics of fermented black garlic (BG) with probiotics. Nine strains of probiotics were tested in media containing 20% BG. Four of the strains grew well in the BG media: Lactobacillus rhamnosus, L. paracasei subsp. paracasei, L. casei, and L. plantarum. These four strains were used to make 10, 20, and 30% BG fermented product, respectively. The number of viable cells, pH, acidity, S-allyl cysteine (SAC) concentration, and nitric oxide (NO) and reactive nitrogen species (ROS) generation in Raw 264.7 macrophage cells were measured. L. plantarum showed the best growth of all the strains in the BG media. The pH of all the samples decreased during fermentation, and the acidity increased acidity. However, they did not differ significantly from the pH and acidity of the control. In all four strains, the SAC content did not differ before and after fermentation. However, the SAC content increased, depending on the BG concentration. NO production was inhibited in the L. rhamnosus inoculation strain compared to the other strains. ROS generation was also significantly inhibited in the L. plantarum inoculation strain compared to the other strains. The results show that the characteristics of BG fermentation products are determined by the fermentation strain. Therefore, fermentation products with particular characteristics can be produced using a single strain or mixed strains.

Gene Expression Analysis of Immune Cell Activation Markers in Extracts of Platycodon grandiflorum Containing Medicinal Herbs (도라지 추출액과 한약재 함유 도라지 추출액에 의한 면역세포 활성 표지유전자 발현 분석)

  • Kang, Shin Ae;Chun, Sung Sik;Kang, Shin Kwon;Chung, Young Chul;Cheon, Eun Woo;Cho, Sang Uk;Jung, Kyung Hwa;Ahn, Soon Cheol;Yu, Hak Sun
    • Journal of Life Science
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    • v.24 no.5
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    • pp.567-572
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    • 2014
  • Extracts of Platycodon grandiflorum have been reported to show anti-inflammatory, antioxidant, anti-metastatic, and hepato-protective effects. This study was designed to evaluate T-cell activation and M1/M2 differential macrophage activation by extracts of P. grandiflorum or P. grandiflorum containing various medicinal herbs. Using real-time RT-PCR, we analyzed expression levels of c-fos, and CD40L (T-cell activation markers) in splenocytes and iNOS, Ym1, and ARG1 in RAW 246.7 cells after treatment of CC (hot water extract of P. grandiflorum), MAEK (hot water extract of P. grandiflorum [82%] and six different plants), and HWAL (hot water extract of P. grandiflorum [7%] and eight different plants. The results showed that MAEK significantly elevated the expression of T-cell activation markers of splenocytes, with the c-fos gene activated more than 10-fold and the CD40L gene activated more than 6-fold. Although CD40L was significantly increased by CC and HWAL, the increase was only about 2-fold. In addition, CC and HWAL did not significantly activate the expression of the c-fos gene. On the other hand, CC elevated the M1 activation marker iNOS, and HWAL elevated the M2 activation marker Ym1 and ARG1 gene expression. In conclusion, MAEK could be used as an immune stimulant because of its ability to activate T cells (elicited c-fos and CD40L gene expression), whereas HWAL could serve as an anti-inflammatory agent because of its differential activation of M2 macrophages.

Inhibitory Efficacy of Dystaenia takeshimana Extract on iNOS, COX-2 Protein and mRNA Expression in Raw 264.7 Cell (Raw 264.7 세포에서 섬바디나물 추출물의 iNOS, COX-2 단백질 및 mRNA 발현 억제 효과)

  • Lee, Jin-Young;Yoo, Dan-Hee;Joo, Da-Hye;Chae, Jung-Woo
    • Microbiology and Biotechnology Letters
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    • v.44 no.4
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    • pp.571-576
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    • 2016
  • In this study, the anti-inflammatory activities of the 80% ethanol extract of Dystaenia takeshimana (DT) were investigated using Raw 264.7 cells treated with lipopolysaccharide (LPS). The effect of DT extract on the production of pro-inflammatory factors (iNOS, COX-2) in LPS-stimulated Raw 264.7 macrophages was examined. The cytotoxic effect of DT extract on macrophage cells (Raw 264.7) was examined by the 3-[4, 5-dimethyl-thiazol-2-yl]-2, 5-diphenyl-tetrazoliumbromide (MTT) assay. Treatment with DT extract showed 100% or more cell viability at the concentration $1,000{\mu}g/ml$. The inhibitory effect of DT extract on protein expression of inducible NOS (iNOS) and cyclooxygenase-2 (COX-2) was measured by western blotting using the concentrations 50, 100, and $500{\mu}g/ml$, with ${\beta}-actin$ used as the positive control. Consequently, the protein expression of iNOS, and COX-2 as observed by western blotting, was decreased by 56%, 61.6%, respectively with $500{\mu}g/ml$ DT extract. Inhibition of iNOS and COX-2 mRNA expression was measured by reverse transcription- polymerase chain reaction (PCR) using DT extract concentrations 50, 100, and $500{\mu}g/ml$, with GAPDH used as a positive control. Consequently, the mRNA expression of iNOS and COX-2 as observed by reverse-transcription-PCR was decreased by 77.9% and 83.3%, respectively at $500{\mu}g/ml$ concentration of DT extract. In conclusion, DT extract may affect inflammatory factors as a potential anti-inflammatory agent.

Effect of ethanol extract of Lotus Rhizome and node of Lotus Rhizome (연근과 우절 에탄올 추출물의 향장효능 검증)

  • Jang, Young-Ah;Park, So-Hyun;Kim, Bo-Ae;Park, Jong-Yi;Jeoung, Young-Ok;Lee, Jin-Tae
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.3
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    • pp.657-665
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    • 2017
  • This study is for checking the possibility of Lotus Rhizome and node of Lotus Rhizome as cosmetic materials. For this we carried out biological active evaluation about anti-oxidant, anti-inflammatory and anti-wrinkle by using ethanol extract of Lotus Rhizome and node of Lotus Rhizome. We extracted Lotus Rhizome and node of Lotus Rhizome with 95% ethanol and then in order to evaluate anti-oxidant activity we treated samples by concentrations (100, 500, 1000) ${\mu}g/mL$ and carried out 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging and The activity of 2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) cation radical scavenging. Also, in order to evaluate effect of anti-wrinkle we carried out evaluation of Elastase inhibitory activity. To evaluate effect of anti-inflammatory we evaluated toxicity of samples through MTT assay with a macrophage (Raw 264.7 cells) and measured nitric oxide production inhibitory activity. As a result, DPPH radical scavenging activity of Lotus Rhizome and node of Lotus Rhizome at $1000{\mu}g/mL$ was 66.7% and 99.5%, respectively and ABTS + radical scavenging activity was 51.2% and 98.3% at the same concentration, respectively. Elastase inhibitory activity results showed that the nodes of the Lotus Rhizom extract excellent anti-wrinkle efficacy than Lotus Rhizom. Node of Lotus Rhizome showed higher anti-wrinkle activity than the positive the control group BHT at $1000{\mu}g/mL$ concentration. According to the result of nitric oxide production inhibitory activity, Lotus Rhizom showed 55.8% effect and nodes of the Lotus Rhizom showed 66.6% effect respectively. This showed that effect of anti-inflammatory was greater in nodes of the Lotus Rhizom extracts. As a result it suggests that Lotus Rhizome and node of Lotus Rhizome extracts can be used as natural substance of cosmetics which are safe in antioxidant, anti-inflammatory, and anti-wrinkle.

The Role of Lymphocyte Compartment and Cytokine in Coal Workers Pneumoconiosis (진폐증환자에서의 임파구 분획 및 싸이토카인의 역할)

  • Lee, Jung-Yeon;Yoo, Kwang-Ha;Ahn, Hae-Ryon;Kim, Sung-Ryul;Lee, Hae-Woon;Ahn, Cheol-Min;Kim, Hyung-Joong
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.3
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    • pp.241-250
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    • 2002
  • Background : Coal-worker's pneumoconiosis(CWP) is characterized by a chronic inflammatory lung reaction associated with macrophage accumulation in the alveolar spaces. CWP is usually divided into two stage : simple pneumoconiosis(SP) where there are a limited number of fibrotic lesions remain limited, with radiological opacities smaller than 1cm and progressive massive fibrosis(PMF), which is characterized by the development of a perifocal extensive fibrotic response of the lung and severe alterations in pulmonary function. In this study, the lymphocyte compartment and cytokine were evaluated by measuring the serum levels in the control, SP and PMF groups. Materials and Methods : The coal workers selected for this study were employees(patients?) of the Tae-Baek and Dong-Hae hospital. All were men, 45-76 years old and the mean duration of their exposure to coal dust was 23.2 years in the lymphocyte compartment and 24.3 years in the cytokine checked group. According to X-ray examination results, the patients were classified into either one of the SP, PMF categories. The normal controls examined were 26-70 years old men. The serum cytokine levels were estimated by using an end point enzyme immunoassay technique. Results : T lymphocyte, helper and suppressor T cells were highly related to pneumoconiosis in this study. A statistically significant decrease in the number of suppressor T lymphocytes was observed in the simple pneumoconiosis patients and at the same time, there was an increase in the lymphocyte index. Howevere, there was no statistically difference in the serum cytokines levels among the SP, PMF and control groups. Conclusion : T lymphocyte, helper T, and suppressor T cells may be highly related to the development of CWP compared to the control group particularly in the early stage of pneumoconiosis. The changes observed in the immunological system in patients with pneumoconiosis may lie at the bottom of the pathogenesis of fibrosis. Further study is needed to evaluate the lymphocyte compartment as a marker for pneumoconiosis development in the early stage.

Anti-inflammatory Activity of Extracts of Hovenia dulcis on Lipopolysaccharides-stimulated RAW264.7 Cells (LPS로 유도된 RAW264.7 대식세포에 대한 헛개나무(Hovenia dulcis) 추출물의 항염증 효과)

  • Woo, Hyun Sim;Lee, Sun Min;Heo, Jeong Doo;Lee, Min-Sung;Kim, Yeong-Su;Kim, Dae Wook
    • Korean Journal of Plant Resources
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    • v.31 no.5
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    • pp.466-477
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    • 2018
  • In this study, the anti-inflammatory activities of the extracts of different parts of Hovenia dulcis such as leaves, stems, and roots were investigated. Among them, the roots extract (RE) showed the most potent suppressive effect against pro-inflammatory mediators in LPS-stimulated mouse macrophage cells. RE induced dose-dependent reduction of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) and concomitantly reduced the production of NO and $PGE_2$. Additionally, pre-treatment with RE significantly suppressed the production of inflammatory cytokines, such as tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$), interleukin $(IL)-1{\beta}$, and IL-6, as well as mRNA levels. Moreover, phosphorylation of mitogen-activated protein kinases (MAPKs) and nuclear translocation of nuclear factor-kappa B (NF-kB) were also strongly attenuated by RE in RAW264.7 cell. Furthermore, RE induced HO-1 expression through nuclear translocation of nuclear factor E2-related factor 2 (Nrf2) and increase HO-1 activity in RAW264.7 macrophages. Therefore, these results indicate that RE strongly inhibits LPS-induced inflammatory responses by blocking NF-kB activation, inhibiting MAPKs phosphorylation, and enhancing HO-1 expression in macrophages, suggesting that RE of H. dulicis and a major component, 27-O-protocatechuoylbetulinic acid could be applied as a valuable natural anti-inflammatory material.

Anti-inflammatory Activity of Crinum asiaticum Linne var. Japonicum Extract and its Application as a Cosmeceutical Ingredient (문주란의 항염효과와 화장료적 특성)

  • Kim, Ki-Ho;Kim, Young-Heui;Kim, Ki-Soo;Park, Sun-Hee;Lee, Soo-Hee;Kim, Young-Jin;Kim, Young-Sil;Kim, Jong-Heon
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.32 no.1 s.55
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    • pp.59-64
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    • 2006
  • Crinum asiaticum Linne var. japonicum has long been used as a rheumatic remedy, an anti-pyretic, an anti-ulcer treatment, and for the alleviation of local pain and fever in Korea and Malaysia. In order to investigate the possibility of Crinum asiaticum Linne var. japonicum extract as a cosmetic ingredient, we measured its anti-inflammatory effect by inhibition of iNOS (inducible nitric oxide synthase), and the release of PGE2, IL-6, and IL-8. HPLC experiment after extraction with 95% ethanol at pH 3.5 showed that Crinum asiaticum Linne var. japonicum was mainly composed of lycorine (up to 1%), a well-known immunosuppressant. The content of lycorine varied depending on the type of tissue analyzed and the extraction method. In anti-inflammatory assay for inhibition of nitric oxide formation on lipopolysaccharide (LPS)- activated mouse macrophage RAW 264.7 cells, the ethanolic extract of Crinum asiaticum showed inhibitory activity of NO production in dose-dependent manner ($IC_{50} = 83.5 {\mu}g/mL$). Additional study by RT-PCR demonstrated that the extract of Crinum asiaticum significantly suppressed the expression of the iNOS gene. Moreover, the extract of Crinum asiaticum did not show my cytotoxicity, but did show cell proliferation effect against LPS ($10{\sim}60%$ increase of tell viability). In an assay to determine inhibition of the $H_2O_2$-activated release of PGE2, IL-6, and IL-8 in human normal fibroblast cell lines, the release of PGE2 and IL-6 was almost completely inhibited above concentrations of 0.05% and 1%, respectively. Moreover, the release of IL-8 was completely inhibited over the entire range of concentrations (> 0.0025%). The result showed that the extract of Crinum asiaticum Linne var. japonicum has sufficient anti-inflammatory effect. There-fore, Crinum asiaticum Linne var. japonicum extract may be useful as an ingredient of cosmetic products.