• Title/Summary/Keyword: cytokine ELISA

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LEVELS OF IL-1 AND TNF-α IN ODONTOGENIC CYST & CYSTIC FLUID (치성낭종과 낭종액에서 IL-1, TNF-α의 농도분포에 관한 연구)

  • Gong, Hyung-Gyu;Park, Dong-Sung;Lim, Sung-Sam
    • Restorative Dentistry and Endodontics
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    • v.24 no.1
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    • pp.49-54
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    • 1999
  • Ko, Lim found some differences in the concentrations of bone resorptive cytokines, especially IL-$1{\alpha}$ and IL-$1{\beta}$ in periapical lesions and inflamed pulps. And they suppose that these differences may be due to the type of cells which produce each cytokine. The purpose of this study was to analyze the human odontogenic cysts & cystic fluid for their contents of IL-$1{\alpha}$, IL-$1{\beta}$ and TNF-$1{\alpha}$ and to compare the concentrations of each cytokine according to the cytokine producing cells. The cystic tissues used in this experiment, were obtained from periapical surgery or cyst enucleation surgery. Cystic fluid was obtained from root canal during routine endodontic therapy(n=5). Cystic tissues were subdivided into two groups, inflammatory radicular cyst group(n=15) and developmental odontogenic keratocyst group(n=3). Normal periapical tissues of extracted third molar(n=5) were also obtained to be used as control group. Each specimen was incubated in 0.5ml homogenizing buffer (0.1mol/L potassium chloride, 0.02mol/L TRIS;pH=7.6) for two hours and then homogenized with glass homogenizer. Each specimen was centrifuged in a microcentrifuge for 3 minutes, and supernatants were extracted. The concentrations of cytokines were measured with R&D ELISA kit. The data were analyzed by Mann-Whitney U test for the differences among the diseases and t test for the correlations among each cytokine. Following results were obtained ; 1. For IL-$1{\alpha}$ and IL-$1{\beta}$, all experimental groups showed significantly higher concentrations of each cytokine than the control group (p<0.05). 2. In radicular cysts, the concentrations of IL-$1{\alpha}$ were higher than IL-$1{\beta}$, but not stastically significant (p>0.05). In odontogenic keratocysts, the concentrations of IL-$1{\alpha}$ were significantly higher than IL-$1{\beta}$ (p<0.05). In cystic fluid, the concentration of IL-$1{\beta}$ was significantly higher than IL-$1{\alpha}$ (p<0.05). 3. Between odontogenic keratocysts and radicular cysts, the concentrations of IL-$1{\alpha}$ were significantly higher in odontogenic keratocysts than in radicular cysts (p<0.05). 4. For TNF-${\alpha}$, only cystic fluid group showed significantly higher concentrations than the control group (p<0.05).

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The Effect of Yangkyuk-Sanhwa-Tang on Cytokine Production in the Patients with Cerebral Infarction (중풍환자에서 양격산화탕에 의한 세포활성 물질 생성의 조절 효과)

  • Lo, Hyun-Soo;Lee, Sang-Min;Bae, Young-Chun;Park, Hye-Sun;Lee, Jae-Heung;Song, Seung-Yun;Yoo, Kwan-Seok;Joo, Jong-Chon
    • Journal of Sasang Constitutional Medicine
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    • v.16 no.1
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    • pp.120-129
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    • 2004
  • Yangkyuk-Sanhwa-Tang(YST) has been widely used as a formula for the Soyangin cerebral infarction (CI) patients according to Sasang constitutional philosophy. Brain cells produce cytokines and chemokines during the inflammatory process after stroke both in animal models and in patients. Previously, regulation of serum cytokine levels by YSThas been observed in individuals at the acute stage of CI disease, but there have not been other scientific investigations on YST. The author investigated the effect of YST on theproduction of various cytokines using peripheral blood mononuclear cells (PBMCs)from the Soyangin (CI) patients, and Soyangin normal group. The cytokine production was analyzed using enzyme-linked immunosorbent assay (ELISA). The amount of interleukin (IL)-1, IL-1, IL-6, IL-8, and tumor necrosis factor (TNF)- in culture supernatant significantly increased in the LPS-treated cells compared with unstimulated-cells (P < 0.05). However, in LPS-stimulated PBMCs, cytokines level in CI patients group was higher than that of normal group. YST (1 mg/ml) significantly inhibited IL-1, IL-1, and IL-8 production in PBMCs stimulated with LPS (about 85% for IL-1, 87% for IL-1, and 53% for IL-8, P < 0.05), but did not significantly inhibit IL-6 and TNF- production in the CI patients group. We also show that YST significantly increased LPS-induced IL-1, IL-6, and TNF- production in the normal group. Thesedata suggest that YST has a regulatoryeffect on the cytokine production, which might explain its beneficial effect in the treatment of CI.

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Effects of Whitmania pigra on the expressions of cytokine genes in human THP-1 cells (수질(水蛭)이 THP-1세포의 사이토카인 유전자 발현에 미치는 영향)

  • Kim, Hyun-Ji;Kim, Gyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.26 no.1
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    • pp.19-34
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    • 2013
  • Objective : This study was performed to evaluate the effect of immune reaction inductive substances such as PMA, LPS, DPE, DNCB and WP, the whitman pigra extracting substance at simultaneously on the translocation of $NF{\kappa}B$ towards to the nucleus and the mRNA expression patterns of various cytokine genes in THP-1 cells, monocytes of human. Methods : To analyze the cytokine genes expressions, the RT-PCR method was used, and measuring TNF-${\alpha}$ that had been secreted during cell culture by the ELISA method. The morphological changes were observed during THP-1 cell by a scanning electron microscope and the quantitative distribution of $NF{\kappa}B$ in the cell that was analyzed through immunocytochemistry and a confocal microscopy. Results : WP showed different influences onto the mRNA expression patterns of cytokine genes with PMA, LPS. DPE and DNCB according to the types of immune inductive substances in the THP-1 cells. Upon treating PMA and DPE on the THP-1 cells at the same time or either additionally treating WP thereon, the movement of $NF{\kappa}B$ increase towards the nucleus from cell cytoplasm was able to be observed. The expressions of IL-$1{\alpha}$ and IFN-${\gamma}$ induced by PMA and PMA+DNCB were suppressed by WP while the expression of TGF-${\beta}$ was promoted. Regarding the secretion pattern of TNF-${\alpha}$ according to the treatment of PMA, its secretion amount was incredibly increased by concurrent treatment of WP, however, in case of co-treatment of WP with PMA and DNCB, it was found that the secretion amount of TNF-${\alpha}$ decreased. Conclusions : In this study, the WP extracting substance was confirmed that it had an influence on expression patterns of cytokine genes according to the actions of a variety kinds of immune reaction inductive substances treated on the THP-1 cells. Especially, WP co-treatment with PMA and DNCB was suppressed the expression of inflammatory cytokines, such as IL-$1{\alpha}$, IFN-${\gamma}$ and TNF-${\alpha}$.

Effects of Extract from Fermented Flower-buds of Panax ginseng C.A. Meyer on Mouse Cytokine IL-6, TNF-α Production (발효 인삼꽃 추출물의 경구 투여가 마우스 사이토카인 IL-6, TNF-α의 생성에 미치는 영향)

  • Jeong, Su-Ji;Kim, Kyoung-Hee;Son, Hwa-Young;Yook, Hong-Sun
    • The Korean Journal of Food And Nutrition
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    • v.27 no.1
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    • pp.43-49
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    • 2014
  • Panax ginseng C.A. Meyer has been used as a traditional medicinal ingredient and the ginseng flower-buds also proved to have good medicinal properties. In this study, in order to enhance immune activities of ginseng flower-bud, the ginseng flower-bud extract was being fermented by Bacillus subtilis KCTC 1022 (BS), Lactobacillus plantarum KCTC3 and Saccharomyces cerevisiae strain CHY1011 (SC). Mice were orally administered daily for two weeks at two different concentrations (100 and 200 mg/kg B.W.). Treatment samples were water extracts of ginseng flower-buds (FD), water extracts of fermented ginseng flower-buds (FM) and controls for saline solution. Cytokine production (IL-6, TNF-${\alpha}$) either stimulated with LPS or not stimulated with LPS was detected by the ELISA assay when using the cytokine kit. Cytokine was statistically increased at supplemented groups with LPS in both the 100 and the 200 mg/kg B.W. and treatment with FM significantly decreased the LPS-induced TNF-${\alpha}$ and IL-6 production more than the treatment with FD. The results of this study may suggest that supplementation with FM increases the immune function by regulating cytokine production capacity for activated macrophages.

Enhancing Effect of Acanthopanax senticosus Extracts on Mouse Spleen and Macrophage Cells Activation (가시오가피 물 추출물에 의한 마우스 비장세포 및 대식세포 활성의 항진효과)

  • Ryu, Hye-Sook
    • The Korean Journal of Food And Nutrition
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    • v.28 no.2
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    • pp.253-257
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    • 2015
  • Acanthopanax senticosus is an herb that has been used as a traditional remedy and medicine source. Its anti-inflammatory and, anti-oxidative effects have been reported in previous studies. This study aimed to investigate the effect of Acanthopanax senticosus water extracts on mouse macrophage cell in vitro. Mouse splenocyte proliferation increased after application of Acanthopanax senticosus water extract supplement of 5, 10, 50, 100, 250, 500, $1,000{\mu}g/mL$ after 48 h pre-treatment with a mitogen (ConA or LPS). The production of cytokines secreted by LPS and non LPS stimulated macrophages was detected by ELISA assay using a cytokine kit. Cytokine production (IL-2, IFN-${\gamma}$, and TNF-${\alpha}$) increased after water extract supplementation. The result of this in vitro study, showed that splenocyte proliferation and cytokine production by activated peritoneal macrophages were increased after Acanthopanax senticosus water extract in the range of $500{\sim}1,000{\mu}L/mL$. Thus, it is suggested that supplementation with Acanthopanax senticosus water extracts may enhance immune function by regulating splenocyte proliferation and enhancing cytokine production by activated macrophage.

Anti-inflammatory and Anti-allergic Effects of Lentinula edodes Extract by UVIrradiation (UV-B 조사에 따른 버섯 추출물의 항염증 및 항알레르기 활성)

  • Hwang, Mi Sun;Pyo, Jaesung;Kim, Hyun Jin;Do, Sun Gil;Song, Il Dae;Kim, Kang Min
    • Journal of Life Science
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    • v.32 no.5
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    • pp.368-374
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    • 2022
  • In this study, the effects of UV irradiation-enhanced ergocalciferol (vitamin D2) content containing Lentinula edodes extract on inflammation and allergic responses were investigated in vitro. The anti-inflammatory and anti-allergic effects of the mushroom extract were tested by estimating the cytokine secretions, such as TNF-α, IL-6, and IL-1β in LPS-activated macrophages (RAW 264.7), or histamine release in PMA and A23187-activated mast cells (RBL-2H3). Under the condition of macrophage activation with LPS, mushroom extract significantly reduced the secretions of pro-inflammatory cytokines, TNF-α and IL-6, and their mRNA expression also matched the observation. The current mushroom extract also significantly reduced the amount of mast cell degranulation-induced histamine secretion from PMA- and A23187-treated mast cells as well as the reduced secretion of IL-4. These results suggest that mushroom extract, which has increased ergocalciferol content by UV irradiation, inhibits the expression of cytokines in inflammation and allergic reactions; therefore, it can be used effectively for the prevention and treatment of inflammatory and allergic diseases.

Effects of Acanthopanax koreanum Extracts on Anticancer Related Cytokine Secretions (섬오가피 추출물의 항암관련 사이토카인 분비활성)

  • Lyu, Su-Yun;Park, Won-Bong
    • YAKHAK HOEJI
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    • v.54 no.4
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    • pp.232-239
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    • 2010
  • Stems and roots of Acanthopanax koreanum Nakai were extracted with water and treated on immune cells in order to determine their immunomodulatory activites. Various Th-1 type cytokines were measured using ELISA including interleukin (IL)-2, IL-12, interferon-gamma (IFN-$gamma$), and tumor necrosis factor-alpha (TNF-$\alpha$) secreted by dendritic cells, T-cells, intestinal epithelial cells, natural killer cells, and macrophages. As a result, there was a significant increase in IL-12 and IFN-$\gamma$, secretion, but there was no change in the secretion of TNF-$alpha$. Additionally T-cells slightly increased the secretion of IL-2, but there was a significant increase of IL-2 in intestinal epithelial cells. Therefore, our results suggest that A. koreanum Nakai may act as an immunomodulator by stimulating the cell-mediated immunity which can help the immune system defend against infections or cancer cells.

Effect of Scutellariae Radix Extract on Serum Cytokines in OVA-induced Asthmatic Mice (황금이 천식모릴 생쥐의 혈청 사이토카인에 미치는 영향)

  • Na, Do-Gyun;Park, Yang-Chun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.4
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    • pp.973-976
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    • 2005
  • The purpose of this research is to examine the effects of Scutellariae Radix(SR) extract on cytokines in ovalbumin (OVA)-induced asthmatic mice. In vivo, C57BL/6 mice were sensitized and handicapped by OVA for 12 weeks. During this experiment, the one group was then treated with SR extract for the later 8 weeks (3 times per week) and analyzed by ELISA. There were significant decreases in IL-4(p<0.05), IL-5(p<0.05), IL-13(p<0.01), histamine(p<0.05) in serum of SR group. IgE also decreased, but was not significant compared with that of control group. The results of this study support a role for SR as an effective treatment for asthma in its experimental success in significantly decreasing inflammation and asthma reactions.

A Study of Serum Cytokines in the Patients with Chronic Hepatitis B Virus Infection (만성 B형 간염 바이러스 감염 환자에서 혈청 Cytokine에 관한 연구)

  • Kim, Byung-Ju;Ma, Jae-Sook;Hwang, Tai-Ju
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.1 no.1
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    • pp.90-99
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    • 1998
  • Purpose: The aim of this study was to clarify the serum cytokine pattern in patients with chronic HBV infection in terms of their clinical state. Methods: Intravenous blood samples were taken from 35 patients who were seropositive for HBsAg for at least 6 months and 7 healthy controls. Samples were initially tested for serum aminotransferases and serologic markers for hepatitis B virus by EIA. Serum levels of interleukin(IL)-2, tumor necrosis factor-alpha (TNF-${\alpha}$), interferon-gamma (IFN-${\gamma}$), IL-4, and IL-10 were measured by ELISA. Results: Among 35 patients, seropositive for HBeAg was 20 and for anti-HBe was 15. The histologic diagnosis of 19 patients underwent liver biopsy were chronic persistent hepatitis (CPH) in 10 and chronic active hepatitis (CAH) in 9. Serum IL-10 level in patients seropositive for HBeAg was significantly higher than that in patients seropositive for anti-HBe (p<0.05). All measured cytokine levels in patients with CAH were higher than those of patients with CPH. High values of all measured cytokines except IL-4 were seen in patients with AST and ALT > 100 U/L. High level of IL-4 was seen in patients with normal aminotransferase levels. Conclusion: These results were thought to indicate that anti-inflammatory Th2-like cytokine (IL-10) production in chronic HBV infection is related to circulating HBeAg rather than activity of hepatitis and that Th1 cytokines seem to be associated with the increasing activity of hepatitis.

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Relation of Interleukin-10 in Bronchoalveolar Lavage Fluid and Airway Inflammation in Bronchial Asthma (기관지천식에서 기관지폐포세척액내 IL-10과 기도염증정도의 연관성)

  • Lee, Sook-Young;Youn, Hung-Gue;Shin, Youn;Lee, Sang-Haak;Kim, Seok-Chan;Kim, Kan-Hyoung;Moon, Hwa-Sik;Song, Jeong-Sup;Park, Sung-Hak
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.1
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    • pp.44-52
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    • 1999
  • Background : Airway infiltration by inflammatory cells, particularly of eosinophils, is one of the characteristic features of asthma. Several mechanisms for the recruitment of eosinophil is focused on the CD4+ T lymphocyte for the preferential production of Th2-c1erived cytokines. Interleukin-10(IL-10) is identified cytokine with potent antiinflammatory activity. This molecule has been shown to inhibit the release of cytokine from inflammatory cells including Th2 cell, and also to inhibit eosinophil survival. We therefore attempted to determine whether decreased synthesis of IL-10 in the lung of bronchial asthma may contribute to inflammation that is characteristics of this dease. Method: Subjects were patients with bronchial asthma(n=23) and normal controls(n=11). IL-10 produced from peripheral mononuclear cell(PBMC) and in bronchoalveolar lavage(BAL) fluid was measured by ELISA method. Degree of bronchial inflammation was assessed by total cell counts and eosinophil percents in BAL fluid, eosinophil infiltration on bronchial biopsy tissue and $PC_{20}$ for methacholine. Results: The IL-10 level produced by PBMC and in BAL fluid from patient with bronchial asthma were not different with normal controls(respectively, $901.6\pm220.4$ pg/ml, $810.9\pm290.8$ pg/ml for PBMC, $24.5\pm9.5$ pg/mL $30.5\pm13.5$ pg/ml for BAL fluid p>0.05). There were significant negative correlation between IL-10 in BAL fluid and eosinophil percents in BAL fluid or degree of eosinophil infiltration in bronchial biopsy (respectively r=-0.522, r=-0.4486 p<0.05). However there was no difference of IL-10 level according to $PC_{20}$ for methacholine. There were no correlation between IL-10 production by PBMC and peripheral blood eosinophil counts or serum eosinophilic cationic protein levels(respectively r=0.1146, r=0.0769 p>0.05). Conclusion: These observation suggest that IL-10 may participate but not acts the crucial role in regulation of the airway inflammation in bronchial asthma.

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