• Title/Summary/Keyword: Cytokine assay

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Inhibitory Effect of Agaricus Mixed Prescription on Metastasis and Tumor Formation

  • Chu Ji Hee;Lim Nak Cheol;Kim Sung Hoon;Ryu Ki Won
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.560-567
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    • 2003
  • Antimetastatic effects of Agaricus mixed prescription (AMP) were studied in the respect of blood-borne metastasis. For this aim, cytotoxicity against various cancer cells and normal cells, Chicken Chorioallantoic membrane (CAM) assay, cancer cell adhesion assay, platelet aggregation assay, pulmonary colonization, life span of S-180 implanted mice, and cytokine release assay were evaluated, respectively. The results were summarized as follows; AMP did not exert any cytotoxicity against all cell lines with IC50 of 25mg/ml on B16BL6. AMP disrupted formation of CAM at 1mg/ml. AMP was suppressive in adhesion assay of B16BL6. AMP also inhibited tumor induced platelet aggregation. In pulmonary colonization assay by B16BL6, the number of colonies in the lungs was significantly decreased in sample group than in control group. In animal study with S-180, the life span of AMP treated group was extended than that of control group. IL-12 was effectively increased in AMP treated group in cytokine release assay. Taken together, AMP can be possibly applied to cancer or metastasis.

Inhibitory Effects of the Ethanol Extract of Ulmus davidiana on Apoptosis Induced by Glucose-glucose Oxidase and Cytokine Production in Cultured Mouse Primary Immune Cells

  • Lee, Jeong-Chae;Lim, Kye-Taek
    • BMB Reports
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    • v.34 no.5
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    • pp.463-471
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    • 2001
  • The bark of Ulmus darvidiana var. japonica Nakai (UDN) has been used for a long time to cure inflammation in oriental medicine. In the present study, two types of extracts, Ulmus water-eluted fraction (UWF) and Ulmus ethanol-eluted fraction (UEF), were prepared from the UDN stem bark, and employed to test the extracts to see if they had anti-oxidative properties against hydroxyl radicals that could alter immune reactivity in mouse immune cells. Deoxyribose assay, DNA nicking assay, and glucose/glucose oxidase assay showed that both fractions had scavenging activity against oxygen free radicals at 50 mg/ml. In addition, hydroxyl radical-mediated apoptosis in mouse thymocytes was not protected by UEF treatment, but the apoptosis was protected by UWF at the same concentration. DNA synthesis and cytokine production that were induced in splenocytes by mitogens (Concanavalin A and lipopolysaccharide) were reduced by the addition of both fractions. These results indicate that both extracts that were prepared from the UDN stem bark have anti-oxidative activities, anti-apoptotic effects, and inhibitory effects on DNA synthesis and cytokine production in mouse immune cell cultures.

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Pycnogenol attenuates the symptoms of immune dysfunction through restoring a cellular antioxidant status in low micronutrient-induced immune deficient mice

  • Lee, Jeongmin;Nam, Da-Eun;Kim, Ok-Kyung;Lee, Myung-Yul
    • Nutrition Research and Practice
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    • v.8 no.5
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    • pp.533-538
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    • 2014
  • BACKGROUND/OBJECTIVES: We investigated the effect of Pycnogenol (Pyc) on survival and immune dysfunction of C57BL/6 mice induced by low micronutrient supplementation. MATERIALS/METHODS: Female C57/BL/6 mice were fed a diet containing 7.5% of the recommended amount of micronutrients for a period of 12 wks (immunological assay) and 18 wks (survival test). For immunological assay, lymphocyte proliferation, cytokine regulation, and hepatic oxidative status were determined. RESLUTS: Pyc supplementation with 50 and $100mg{\cdot}kg^{-1}{\cdot}bw{\cdot}d^{-1}$ resulted in partial extension of the median survival time. Pyc supplementation led to increased T and B cell response against mitogens and recovery of an abnormal shift of cytokine pattern designated by the decreased secretion of Th1 cytokine and increased secretion of Th2 cytokine. Hepatic vitamin E level was significantly decreased by micronutrient deficiency, in accordance with increased hepatic lipid peroxidation level. However, Pyc supplementation resulted in a dose-dependent reduction of hepatic lipid peroxidation, which may result from restoration of hepatic vitamin E level. CONCLUSION: Findings of this study suggest that Pyc supplementation ameliorates premature death by restoring immune dysfunction, such as increasing lymphocyte proliferation and regulation of cytokine release from helper T cells, which may result from the antioxidative ability of Pyc.

An Experimental Study on the Anti-inflammatory Effects of Cheonggisan Extract in Allergic Late Inflammation (알레르기 후기 반응 염증 억제효과에 관한 청기산(淸肌散)의 실험적 연구)

  • Jo, Suk-Yong;Kang, Min-Seo;Kim, Yong-Min;Kim, Hee-Taek
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.29 no.3
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    • pp.95-105
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    • 2016
  • Objectives : Allergic diseases have a various symptoms of hyperresponsiveness and recently hyperresponsive reaction in the chronic phase is reported as the important mechanisms. Cheonggisan(CGS) is used in oriental clinics for curing various skin diseases due to effect of controlling of pruritus. There have been studies on the anti-allergic effect and anti-inflammatory effect of CGS, but there had no study of anti-allergic effects in allergic late inflammation of CGS, so we aimed to find out the effects of CGS in allergic late inflammation in our study.Methods : To investigate the anti-allergy effect and anti-inflammatory effect of CGS, RAW 264.7 macrophage cells and CSG water-extracts were used. Cytotoxic effect of CSG was examined by MTT assay, an oxidative product of NO was measured in the culture medium by the Griess reagent assay. The level of prostaglandin E2(PGE2) was measured by competitive enzyme-linked immunoassay. Cytokine(PGE2, IL-1β, IL-6, TNF-α) was measured by Bio-Plex suspension assay system and quantitative multiplexed cytokine/chemokine assay.Results : We investigated that there was no cytotoxic effect of CGS water-extract at any levels of concentration on RAW 264.7 macrophage cells by MTT assay. CGS water-extracts significantly suppressed the levels of the inflammatory mediators such as NO and PGE2, cytokine of IL-1β, TNF-α at the level of 400 ㎍/㎖ CGS concentration. But there was no significant effect on IL-6 production suppression.Conclusions : These results suggest that CSG water-extract has and anti-inflammatory effects in allergic reaction. These properties may contribute to the allergic diseases and inflammatory related disease care.

Effect of Theobroma cacao L. Extracts on Mouse Spleen Immune Cells Activation (카카오 열수 추출물 투여의 마우스 면역세포 활성효과)

  • Ryu, Hye-Sook
    • The Korean Journal of Food And Nutrition
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    • v.33 no.6
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    • pp.732-736
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    • 2020
  • Theobroma cacao L., a fruit of cacao trees, is a perennial plant, which belongs to Sterculiaceae, and is native to the Amazon in South Africa. It also has been known for its various biologically active effects, such as anti-oxidation, anti-cancer, and anti-bacterial. The spleen cell proliferations of mice were measured at 48 hours after treatment of Theobroma cacao L. water extracts in seven concentrations(0, 5, 10, 50, 100, 250, 500 and 1,000 ㎍/mL) an ELISA assay. The production of cytokine (IL-1β, TNF-α, IFN-γ), is secreted by macrophages stimulated with LPS, was detected by ELISA assay using the cytokine kit. From the results of in vitro study, both splenocytes and cytokine production activated by peritoneal macrophages have increased when water extracts were supplemented in the range between 250 and 500 ㎍/mL concentration. Notably, splenocytes production has a signigicant proliferation at 500 ㎍/mL concentration. The result from this research suggests that supplementation with Theobroma cacao L. water extracts may enhance the immune function by stimulating the splenocyte proliferation and improving the cytokine production activating macrophage in vitro.

Effect of Injin Butanol Fraction with Thin Layer Chromatography on Fas-mediated Apoptosis (인진butanol 분획의 TLC추출성분이 Fas-mediated Apoptosis에 미치는 영향)

  • 박용진;김영철;이장훈;우흥정
    • The Journal of Korean Medicine
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    • v.23 no.2
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    • pp.57-69
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    • 2002
  • Objective: The aim of this study is to investigate the effect of Injin butanol fractions with Thin Layer Chromatography on Fas-mediated Apoptosis. Method: Injin-butanol fraction separated by TLC. MIT assay, cell cycle analysis, Caspase-3 protease assay, DNA fragmentation assay and quantitative RT-PCR were performed to evaluate the effects of TLC extraction of lnjin-butanol fraction on cell viability, cell cycle progression and apoptosis. Results: Scopoletin, luteolin, apigenin and unknown powder was isolated by TLC. Fas-mediated apoptosis analysis shows that scopoletin has inhibiting function on apoptosis. Caspase- 3 protease assay analysis shows that scopoletin inhibits activity of caspase-3. Quantitative RT-PCR analysis shows that no activity on caspase-3, but apoptosis inhibition cytokine -Bcl-2- is activated, and apoptosis activating cytokine -Bax- is unactivated. Conclusion: These results show that each fraction of Injin-butanol TLC extraction, especially scopoletin, acts as a protective function on liver cell viability, and inhibitory function on apoptosis. (J Korean Oriental Moo 2002;23(2):57-69)

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Effects of Gardeniae Fructus on Cytokines in Mouse Macrophage (치자(梔子)가 대식세포의 Cytokine에 미치는 영향)

  • Cha, Ji-Hea;Lim, Eun-Mee
    • The Journal of Korean Obstetrics and Gynecology
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    • v.27 no.1
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    • pp.1-16
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    • 2014
  • Objectives: The purpose of this study was to investigate the effects of Gardeniae Fructus Water Extract (GF) on the production of inflammatory mediators in RAW 264.7 cell treated with lipopolysaccharide (LPS). Methods: Gradeniae Fructus was extracted with distilled water (2,000 ml) for 2 hours. In order to evaluate cytotoxicity of GF, 3 - (4,5-dimethylthiazol-2-yl) - 2,5 - diphenyltetrazolium bromide (MTT) assay was performed. To investigate antiinflammatory effects, the concentration of nitric oxide (NO) was measured with No assay, calcium (Ca) was measured with Fluo-4 Ca assay, and cytokine was measured by Bio-Plex cytokine assay in RAW 264.7 cell. And when p-value is below 0.05, it is judged to have the significant difference statistically. Results: 1. GF did not show any cytotoxicity. 2. GF suppressed the production of NO and Ca at the concentration of 25, 50, 100 and $200{\mu}g/ml$. 3. GF suppressed the production of interleukin (IL)-$1{\beta}$, IL-10, IL-12p40, macrophage-colony stimulating factor (M-CSF), macrophage inflammatory protein (MIP)-$1{\beta}$ and keratinocyte chemoattractant(KC) at the concentration of 25, 50, 100 and $200{\mu}g/ml$. 4. GF suppressed the production of vascular endothelial growth factor (VEGF), granulocyte-colony stimulating factor (G-CSF) and monocyte cheomattractant protein (MCP)-1 at the concentration of 25, 50 and $100{\mu}g/ml$. 5. GF suppressed the production of granulocyte macrophage-colony stimulating factor (GM-CSF) and regulated on activation, normal T cell expressed and secreted (RANTES) at the concentration of 25 and $50{\mu}g/ml$. 6. GF suppressed the production of MIP-2 at the concentration of 50 and $100{\mu}g/ml$, and tumor necrosis factor (TNF)-${\alpha}$ at the concentration of 50 and $200{\mu}g/ml$. Conclusions: These results suggest that GF has anti-inflammatory effect and immuno-modulating activity.

Assessment of cytokine producing cells to discriminate between allergen and irritant in the local lymph node assay

  • Lee, Jong-Kwon;Byun, Jung-A;Park, Seung-Hee;Kim, Hyung-Soo;Park, Jae-Hyun;Ryu, Mi-Hyun;Park, Han-Jin;Juno H. Eom;Oh, Hye-Young
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2003.10b
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    • pp.124-124
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    • 2003
  • A murine local lymph node assay(LLNA) has been developed as an alternative method to guinea pig maximization test for contact sensitization potential. This study was carried out to investigate the potential use of cytokine producing cells to discriminate between allergens and irritant in the LLNA.(omitted)

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Study on the Anti-Inflammatory Activity and Mechanism of Medicinal Plants Used in the Treatment of Arthritis (관절염 치료에 사용되는 한약재들의 항 염증 활성과 기전에 관한 연구)

  • Kim, Yoo-Hyun;Park, Ho
    • Korean Journal of Clinical Laboratory Science
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    • v.48 no.3
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    • pp.176-182
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    • 2016
  • When inflammatory reaction is in progress, the macrophages release inflammatory cytokines, such as interleukin-6 (IL-6), tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), and product inflammatory mediators, including inducible nitric oxide synthase (iNOS) and prostaglandin E2 (PGE2). We conducted this study to evaluate the anti-inflammatory efficacy on each water extract of Acanthopanacis cortex, Achyranthes radix, and Eucommiae cortex, and to investigate whether they inhibit the expression of pro-inflammatory cytokine. Acanthopanacis cortex, Achyranthes radix, and Eucommiae cortex were extracted with water and freeze-dried. Acanthoside D, 20-hydroxyecdysone, and pinoresinol diglucoside as an index material were analyzed by high-performance liquid chromatography (HPLC) to ensure that the components of each extracts were extracted well. RAW 264.7 cell line, stimulated with lipopolysaccharide (LPS) to cause an inflammatory response, was treated with each water extract at various concentrations to determine the anti-inflammatory efficacy. Then, the anti-inflammatory efficacy was confirmed by a nitric oxide (NO) assay, and the mRNA expression levels of pro-inflammatory cytokines were measured by real time PCR. As a result, the indicator materials were detected from each extract, and Acanthopanacis cortex water extract (ACWE) and Achyranthes radix water extract (ARWE) were shown to have a high activity than Eucommiae cortex water extract (ECWE) in NO assay. In Korea, traditionally it prescribed a combination of medicinal herbs. This study confirmed the anti-inflammatory response of these medicinal plants in arthritis and its synergistic effect when used in combination with western medicine.

Naegleria fowleri Lysate Induces Strong Cytopathic Effects and Pro-inflammatory Cytokine Release in Rat Microglial Cells

  • Lee, Yang-Jin;Park, Chang-Eun;Kim, Jong-Hyun;Sohn, Hae-Jin;Lee, Jin-Young;Jung, Suk-Yul;Shin, Ho-Joon
    • Parasites, Hosts and Diseases
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    • v.49 no.3
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    • pp.285-290
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    • 2011
  • Naegleria fowleri, a ubiquitous free-living ameba, causes fatal primary amebic meningoencephalitis in humans. N. fowleri trophozoites are known to induce cytopathic changes upon contact with microglial cells, including necrotic and apoptotic cell death and pro-inflammatory cytokine release. In this study, we treated rat microglial cells with amebic lysate to probe contact-independent mechanisms for cytotoxicity, determining through a combination of light microscopy and scanning and transmission electron microscopy whether N. fowleri lysate could effect on both necrosis and apoptosis on microglia in a time- as well as dose-dependent fashion. A $^{51}Cr$ release assay demonstrated pronounced lysate induction of cytotoxicity (71.5%) toward microglial cells by 24 hr after its addition to cultures. In an assay of pro-inflammatory cytokine release, microglial cells treated with N. fowleri lysate produced TNF-${\alpha}$, IL-6, and IL-$1{\beta}$, though generation of the former 2 cytokines was reduced with time, and that of the last increased throughout the experimental period. In summary, N. fowleri lysate exerted strong cytopathic effects on microglial cells, and elicited pro-inflammatory cytokine release as a primary immune response.