• Title/Summary/Keyword: cytokine ELISA

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Effect of Ochnaflavone as An Immunoadjuvant (Ochnaflavone의 면역보조제 효과)

  • Park, Minjoo;Rhew, Ki Yon;Han, Yongmoon
    • YAKHAK HOEJI
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    • v.56 no.6
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    • pp.366-371
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    • 2012
  • In this present study, we determined whether or not there is an immunoadjuvant effect of ochnaflavone, a biflavone isolated from Lonicera japonica. As an antigenic source, the cell wall (CACW) of Candida albicans, a fungal pathogen, was used. CACW consists of 95% carbohydrate (mannan). In the experiments, BALB/c mice were immunized with emersion forms of CACW combined with or without ochnaflavone (Och) in the presence of IFA containing mineral oil or CACW alone. Then, the amounts of antisera collected from these mice groups were measured by the ELISA method. Data from these experiments showed that CACW combined with Och (CACW/Och/IFA) provoked the production of antisera app. 2.2 or 5 times more than the corresponding CACW/IFA or CACW alone (CACW/DPBS), respectively, in mice (P<0.05). We further examined the immune response type induced by Och. Analysis of the values of the IgG1/IgG2a ratios obtained from IgG isotyping revealed that Och induced Th2-immunity more dominantly than Th1. This finding was confirmed by cytokine profile. CACW/Och/IFA formulation induced IL-4 (Th2-type cytokine) more than IFN${\gamma}$ (Th1-type cytokine) as compared with CACW/IFA and CACW/DPBS formulations (P<0.05). All data combined, Och appears to have an immunoadjuvant activity that may convert Th1 immunity into Th2 immunity.

Interleukin-5 Inhibition Assay of the Oriental Materia Medica Treatment by Sandwich ELISA on Mouse Splenocytes (ELISA를 이용한 IL-5 분비조절 한약물 Screening)

  • Park, Ki-Bok;Jung, Sung-Ki;Jung, Hee-Jae
    • The Journal of Internal Korean Medicine
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    • v.30 no.3
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    • pp.582-593
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    • 2009
  • Background and Objective : Allergy is defined as an altered reactivity to an antigen, which can result in pathologic reactions upon subsequent exposure to that particular antigen. This study was to evaluate the effect of oriental materia medica on IL-5 secretion in the mouse spleen cell. Methods : We used the splenocytes of mouse 8 weeks after its birth, and then cultivated those into the 2 experimental groups and a control group for 48 hours. The culture media of the experimental groups were made of $1{\mu}g/ml$, $10{\mu}g/ml$ oriental materia medica, representative. And the culture media of control group was given no oriental materia medica. Then, we assayed the quantity of cytokine-expression by the sandwich ELISA. The quantities of cytokine-expression of the experimental groups were compared with that of the control group, which was standardized. These methods were used for all of the oriental materia medica treated. Results : Some oriental materia medica inhibit the secretion of IL-5 in both $1{\mu}g/ml$ and $10{\mu}g/ml$ culture media. These were Acori Rhizoma, Luffae Fasciculus Vascularis, Amomi Rotundi Fructus, Schisandrae Fructus, Biotae Semen, Clematis armandii, Dioscoreae Sativa Rhizoma, Coicis Semen, Sophorae Flos, Oroxyli Semen, Aurantii Semen, Pini Nodi Lignum. Conclusion : This study indicates that some oriental materia medica inhibit the secretion of IL-5 and are beneficial for allergic disease.

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Protective Effects and Mechanisms of Salvia Miltiorrhizae Radix Extract on Endotoxin Shock (패혈증 쇼크에서 단삼 추출물의 억제 효과와 그 기전에 관한 연구)

  • Shin, Sang-Ho;Yun, Jeong-Min;Ko, Woo-Shin;Yoon, Hwa-Jung
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.29 no.3
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    • pp.14-26
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    • 2016
  • Objectives : This study was performed to investigate the protective effects and mechanisms of Salvia miltiorrhizae Radix extract (SME) on endotoxin shock.Methods : We used two models; LPS-induced sepsis model for in vivo model, and murine peritoneal macrophages responses for in vitro. SME was administrated orally to mice. After 1 hr, LPS was injected intraperitoneally. Survival rate was checked each time per 12 hr for 5 days. Mice were sacrificed 3 hr after LPS injection, then blood samples and organs were harvested. Cytokines secretion was measured by ELISA. Organs tissues were observed with microscope. Murine peritoneal macrophages were cultured for 1 hr either in a medium alone or in a medium that contained SME, as indicated. Then, the cells were treated with LPS for 24 hr. mRNA levels of cytokines were measured by real-time RT-PCR. Cytokine levels in the supernatants were measured by ELISA. The amount of nitrite was measured by using the Griess method to evaluate NO production. The cell lysates were analysed by Western blotting using antibodies for iNOS and β-actin was used as an internal control to monitor equal protein loading.Results : SME improverd the survival rate of mice model. SME inhibited the secretion of inflammatory cytokines and organs damages on Endotoxin Shock model. SME suppressed cytokine expression, cytokine secretion,NO production, iNOS expression in LPS-induced murine peritoneal macrophages.Conclusions : The results suggest that SME has protective effects on endotoxin shock through suppression of inflammatory cytokines, organ damages, NO production and so on.

Inhibitory Effects of Onchungeum on Cytokine Production from Phytohaemagglutinin-stimulated Peripheral blood Mononuclear cells of Behcets Patients (Behcet 환자의 단구세포에서 온청음의 Cytokine 분필억제 효과)

  • Lim Hong Jin;Hwang Choong Yeon;Chiang Hsueh Chuan;Kim Nam Kwon;Kwon Il Ho
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.4
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    • pp.768-773
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    • 2002
  • Behcets disease is a systemic inflammatory disorder. The etiology and pathogenesis of Behcets disease has yet been fully elucidated but might involve immune dysfunction. Cytokines involved in the regulation of inflammatory reactions and immune responses may play a role in the pathogenesis of Behcets disease (BD). Onchungeum is an Oriental herbal medication, which has been successfully used in Korea for the treatment of BD. This report describes modulation effects of Onchungeum on cytokine production from phytohaemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC) of Behcets patients by ELISA. Onchungeum significantly inhibited the production of pro-inflammatory cytokines. TNF-α and IL-1β, compared to absence of Onchungeum (by 52.3 1.4 % inhibition for TNF-α and 113.5 3.3 % for IL-1β, p < 0.001). Onchungeum also inhibited the production of IFN-γ, immunoregulatory Th1 cytokine, by 89.4 0.8 % (p < 0.001). The inhibitory effects of Onchungeum on cytokine production showed dose-dependent manner, and the pre-treatment of 1 mg/ml Onchungeum had better effects than immunosuppressive drug for treatment of BD, cyclosporin A. Our results suggest that Onchungeum treatment for Behcets disease patients may have pharmacologic activities and abilities of regulation of immune and inflammatory responses by cytokine modulation.

Cytokine Gene Expression of Peritoneal Tissues in Response to Mixed Infection of Bacteroides fragilis and Escherichia coli (Bacteroides fragilis와 대장균의 혼합 감염에 의한 복강 조직의 Proinflammatory Cytokine 유전자 발현 조절)

  • Kim, Jung-Mogg;Kim, Young-Jeon;Park, Hwon-Kyum;Cho, Yang-Ja
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.1
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    • pp.41-48
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    • 2000
  • Bacteroides fragilis and Escherichia coli, normal colonic inhabitants, are the most frequently isolated bacteria in infected tissues, particularly in intraabdominal abscesses. This study was designed to determine whether enteric bacteria may alter the B. fragilis-induced expression of pro inflammatory cytokines in mouse peritoneal tissue (MPT). After C57BL/6 mice were inoculated with abscess-forming mixture containing B. fragilis in the presence or absence of E. coli, RNA was extracted from MPT. Expression of interleukin (IL)-$1{\alpha}$ and tumor necrosis factor $(TNF){\alpha}$ mRNA was assessed using RT-PCR and standard RNA. Each cytokine protein was also measured by ELISA. The co-inoculation of E. coli into mouse peritoneal cavity advanced the onset of abscess development by B. fragilis infection. When mouse was co-infected with E. coli and B. fragilis intraperitoneally, there was a synergistic increase in the expression of IL-$1{\alpha}$ and $TNF{\alpha}$ mRNA in MPT and this was paralleled by increased cytokine protein secretion. Mixed inoculation of heat-killed E. coli and B. fragilis did not cause a synergistic increase in those cytokine mRNA expression. These results suggest that enteric bacteria may significantly affect proinflammatory cytokine signal produced by host peritoneal cavity in response to B. fragilis infection.

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Effect of Lipopolysaccharide (LPS) on Mouse Model of Steroid-Induced Avascular Necrosis in the Femoral Head (ANFH)

  • Ryoo, Soyoon;Lee, Sukha;Jo, Seunghyun;Lee, Siyoung;Kwak, Areum;Kim, Eunsom;Lee, Jongho;Hong, Jaewoo;Jhun, Hyunjhung;Lee, Youngmin;Sobti, Anshul Shyam;Kim, Soohyun;Oh, Kwang-Jun
    • Journal of Microbiology and Biotechnology
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    • v.24 no.3
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    • pp.394-400
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    • 2014
  • Avascular necrosis of the femoral head (ANFH) is commonly observed in patients treated with excessive glucocorticoid (GC). Single administration of lipopolysaccharide (LPS) has shown to induce immune stimulatory factors. However, the effect of repeated administration of LPS on GC-induced ANFH has not been studied. Thus, the purpose of this study was (i) to examine the cytokine profile induced by repeated LPS administrations and (ii) to test the effect of repeated LPS treatments on GC-induced ANFH. A mouse necrosis model of ANFH was designed by chronic GC administration with co-treatment of LPS. Mice body weights in the LPS/prednisolone (PDN) co-treated group were lower than that of the untreated control group, but spleen weights were greater than the control group. The levels of IL-6, $TNF{\alpha}$, and IL-33 in the liver and spleen of the LPS/PDN group were lower than the untreated control group, whereas $TNF{\alpha}$ level in the femoral head of the LPS/PDN group increased. Collectively, the effect of repeated LPS on the pathogenesis of GC-induced ANFH was associated with the $TNF{\alpha}$ level in the femoral head, but the pathogenesis did not correspond to cytokine levels in immune tissues.

THE EFFECT OF NATURAL EXTRACTS ON CELL GROWTH AND CYTOKINE PRODUCTION (생약 추출물이 세포성장 및 cytokine 생산에 미치는 영향)

  • Ryu, In-Cheol;Son, Seong-Heui;Chung, Chong-Pyoung;Bae, Ki-Hwan
    • Journal of Periodontal and Implant Science
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    • v.23 no.1
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    • pp.37-47
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    • 1993
  • The native connective tissue attachment of the periodontium is known to be a complex consisting of gingival fibroblasts, periodontal ligament cells, gingival epithelial cells, cementum, alveolar bone and extensive extracellular matrix (collagen, glycoprotein and proteoglycans). The purpose of this study was to evaluate the effects of natural extracts on DNA, collagen and protein synthesis and inhibition of cytokine production in the gingival and periodontal ligament fibroblasts and gingival epithelial cells. Healthy gingival tissue was obtained from orthodontic treatment patients, and gingival epithelial cells, gingival fibroblasts and periodontal ligament cells were isolated and cultured from the samples. After treated with Ginseng protein, Pluronic F-68, Scutellariae Radix, centella asiatica, PDGF, IGF, DNA synthesis, total protein and collagen synthesis, and cytokine production of gingival epithelial cell, gingival fibroblast and periodontal ligamentcells were measured. MTT method for DNA synthesis, Peterkofsky and Dingerman method for total protein and collagen synthesis, and IL-1 ELISA kit for cytokine production were used. The proliferation of epithelial cells was enhanced in Centella asiatica, Ginseng protein, Pluronic F-68 and Scutellariae Radix. The activities of PDL cells were increased in PDGF, IGF, and Pluronic F-68. Higher collagen synthesis was observed in Scutellariae Radix and total protein synthesis was increased in Scutellariae Radix and PDGF. The inhibitory effects on IL-1, IL-6, $TNF-{\alpha}$ were observed in all exrracts.

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The Effect of Mineral Trioxide Aggregate on the Production of Growth Factors and Cytokine by Human Periodontal Ligament Fibroblasts (Mineral trioxide aggregate (MTA)가 치주인대 섬유아세포에서 분비되는 cytokine과 성장인자 TGF-β1, FGF-2 발현에 미치는 영향)

  • Kwon, Ji-Yoon;Lim, Sung-Sam;Baek, Seung-Ho;Bae, Kwang-Shik;Kang, Myung-Hoe;Lee, Woo-Cheol
    • Restorative Dentistry and Endodontics
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    • v.32 no.3
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    • pp.191-197
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    • 2007
  • Mineral trioxide aggregate (MTA) would influence healing of periapical tissues by modulating the production of growth factors and cytokines from PDL fibroblasts, however, the studies are insufficient. Therefore, the purpose of this study was to monitor the expression of transforming growth factor-beta1 $(TGF-\beta1)$, fibroblast growth factor-2 (FGF-2), and interleukin-6 (IL-6) from PDL fibroblasts in the presence of MTA. The human PDL fibroblasts were seeded onto the set MTA or IRM at a level of $1\times10^5$ cells per unit well, and further incubated for 6, 12, 24, and 48 hours. The levels of $TGF-\beta1$, FGF-2 and IL-6 from the supernatant were measured by enzyme-linked immunosorbent assay (ELISA) The data were analyzed using one-way ANOVA. The level of $TGF-\beta1$ was down-reg ulated when the cells were grown in the presence of MTA except at 6 hours. The levels of FGF-2 release were significantly suppressed when PDL fibroblasts were grown in the presence of MTA or IRM at all time intervals (p < 0.05). The expressions of IL-6 from MTA treated co)Is were comparable to those of untreated control cells throughout the observation periods. We presume that this material inhibits the stimulatory function of growth factors on granulation tissue formation and in turn, it promotes the healing process modulated by other bone-remodeling cells.

The effect of Astragali Radix Ethanol extract on Murine CD4 T cells′ Cytokine Profiles in vitro

  • Hee Kang;Bae Hyun Su;Ahn Kyoo Seok
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.5
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    • pp.1330-1334
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    • 2003
  • Astragali Radix(AR), one of the strong tonic herbs, is known to improve immunological responses in mice and human. In this study, AR's ai-reinforcing effect was examined in the context of CD4/sup +/ T cells' TCR/CD3 induced activation responses. In order to evaluate the direct effect of AR on helper T cells, CD4/sup +/ T cells are isolated using magnetic bead and their proliferation and CD69 expression in AR treated medium were assessed with anti-CD3/anti-CD28 activation for 48h. CD4 T cells' proliferation was slightly increased but there was little effect on CD69 expression. RT PCR and ELISA equally demonstrated that IL-2 and IL-4 production was increased but IFN-ν was down-regulated. This shows AR ethanol extract favors Th2 cytokine profile under neutral conditions.

Response of fetal rat calvarial cells on mineral trioxide aggregate after IL-$1{\beta}$ stimulation (IL-$1{\beta}$ 처리 백서 두개관 세포의 mineral trioxide aggregate에 대한 반응)

  • Lee, Sool-Heon;Park, Ji-Il;Kim, Young-Joon
    • Journal of Periodontal and Implant Science
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    • v.39 no.3
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    • pp.359-365
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    • 2009
  • Purpose: The purpose of this study was to investigate the ability of Mineral trioxide aggregate(MTA) to support osteoclastic differentiation from fetal rat calvarial cell. Methods: In this study, response of IL-6, RANKL, and OPG in fetal rat calvarial cells stimulated with IL-$1{\beta}$ on MTA was evaluated by ELISA and RT-PCR. Results: The results were as follows; there was no significant difference between glass and MTA at 5days. In ELISA analysis, Glass group and MTA group showed similar IL-6 expression, Glass+IL-$1{\beta}$ group and MTA+IL-$1{\beta}$ group showed similar IL-6 expression. In RT-PCR analysis, Glass group and MTA group showed similar IL-6, RANKL, OPG mRNA expression, MTA+IL-$1{\beta}$ group and Glass+IL-$1{\beta}$ group showed 3 fold increase of IL-6 and RNAKL mRNA expression when compared with MTA group. All groups showed similar OPG mRNA expression. Conclusions: MTA does not suppress cell proliferation and increase the proinflammatory cytokine that induce osteoclastogenesis. Thus, MTA is biocompatible material that could be used in various clinical conditions.