• Title/Summary/Keyword: cytokine ELISA

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Macrophage activation by glycoprotein isolated from Dioscorea batatas

  • Huong, Pham Thi Thu;Jeon, Young-Jin
    • Toxicological Research
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    • v.27 no.3
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    • pp.167-172
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    • 2011
  • We demonstrate that glycoprotein isolated from Dioscorea batatas (GDB) activates macrophage function. Analysis of the infiltration of macrophages into peritoneal cavity showed GDB treatment significantly increased the recruitment of macrophages into the peritoneal cavity. In order to further confirm and investigate the mechanism of GDB on macrophage activation, we analyzed the effects of GDB on the cytokine expression including IL-$1{\beta}$, TNF-${\alpha}$, and IL-6 in mouse peritoneal macrophages. GDB increased the expression of IL-$1{\beta}$, TNF-${\alpha}$, and IL-6. Cytokine induction by GDB was further confirmed by RT-PCR and ELISA in mouse macrophage cell line, RAW264.7 cells. Treatment of RAW264.7 cells with GDB produced strong induction of NF-${\kappa}B$ DNA binding and MAPK phosphorylation, markers for macrophage activation and important factors for cytokine gene expression. Collectively, this series of experiments indicates that GDB stimulates macrophage activation.

The role of Interleukin-6 and Interleukin-10 in human pulpal inflammation (성인 치수염에서의 INTERLEUKIN-6와 INTERLEUKIN-10의 작용에 관한 연구)

  • Park, Han-Soo
    • Restorative Dentistry and Endodontics
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    • v.27 no.5
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    • pp.515-520
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    • 2002
  • 연구목적 : Cytokine은 유해 미생물에 대한 숙주의 방어기전으로서의 염증반응에서 숙주세포 상호간의 작용을 매개해 주는 역할을 하며, 치수조직에서도 그 존재가 확인된 바 있다. Interleukin-6와 Interleukin-10은 염증의 초기에 작용하는 cytokine으로 알려져 있으나, 치수 및 치근단 질환에서의 역할과 상호작용에 대해서는 잘 알려져 있지 않다. 본 연구에서는 성인의 치수염이 있는 치아에서 Interleukin-6와 Interleukin-10의 농도를 측정하고 이를 정상 치수와 비교함으로써 이들의 치수염에서의 작용을 연구하는 것을 목적으로 하였다. 방법 : 총 60개의 성인 치아들을 대상으로 하였다. 치수염으로 진단된 치아들을 실험군으로 하였고, 정상 치수를 가진 치아들을 대조군으로 하였다. 발치한 치아에서 치수조직을 적출하였다. ELISA를 사용하여 적출된 치수조직 내의 Interleukin-6와 Interleukin-10의 양을 측정하였으며, 그 결과를 Mann-Whitney rank sum test를 사용하여 통계학적 유의성을 검증하였다. 조직학적 검사를 위해서는 발치된 치아에서 치수조직을 적출하여 헤마톡실린-에오신 염색을 시행한 후 관찰하였다. 결과 : 1. Interleukin-6의 농도는 실험군에서 대조군보다 유의하게 높게 나타났다(p<0.05). 2. Interleukin-10의 농도는 실험군에서 대조군보다 유의하게 높게 나타났다(p<0.05). 3. 조직학적 관찰 결과 실험군에서 림프구의 침윤과 부분적인 조직의 괴사 등 염증반응의 양상을 관찰할 수 있었다.

Effect of Plantago asiatica L. Water Extracts on Mice Spleen and Cytokine Cells Activation (질경이 열수 추출 투여의 마이스 사이토카인 및 비장세포 증식 효과)

  • Ryu, Hye-Sook
    • The Korean Journal of Food And Nutrition
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    • v.30 no.3
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    • pp.510-514
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    • 2017
  • Plantago asiatica L., observed frequently in East Asia, is a known herb used in traditional medical remedies several studies report that P. asiatica L has anti-inflammatory and antioxidant effects. To determine the production of cytokines (IL-2, $IFN-{\gamma}$, and $TNF-{\alpha}$) induced by lipopolysaccharide (LPS) and non-LPS-stimulated macrophages, an ELISA assay was conducted using cytokine kits. Mice splenocytes were cultured for 48 h with various concentrations of P. asiatica L. (5, 10, 50, 100, 250, 500, and $1,000{\mu}g/mL$) or with mitogens (ConA or LPS). P. asiatica L. increased the proliferation of mice splenocytes, especially under the condition of its concentration ranging from 250 to $1,000{\mu}g/mL$. In addition, Plantago asiatica L. notably induced cytokine production of (IL-2, $IFN-{\gamma}$, and $TNF-{\alpha}$) at its concentration of $250{\sim}500{\mu}g/mL$. These results suggest that supplementation with P. asiatica L. water extracts may play a potential role in enhancing immune function by mediating splenocyte proliferation and cytokine production through its anti-inflammatory activit.

Ulmus Macrocarpa Water Extract Prolongs Splenocyte Life Span (Ulmus Macrocarpa 열수 추출물에 의한 비장세포 수명 연장)

  • Kang, Kyung-Hwa;Hyun, Sook Kyung;Hwang, Hye Jin;Kim, Byoung Woo;Kim, Cheol Min;Chung, Kyung Tae;Lee, Jong-Hwan
    • Journal of Life Science
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    • v.25 no.10
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    • pp.1176-1183
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    • 2015
  • Ulmus macrocarpa has been used in Korean medicinal food material to physical disorder or tonic material. The purpose of the present study was to evaluate splenocyte life span expansion effects of Ulmus macrocarpa water extract (UMWE) in general cell culture condition. Splenocytes were handled in the presence of 100 μg/ml UMWE for several different time conditions. Live cells were detected with Hoechst 33,342 dye and cell survival molecules were identified through Western blot. Changes in level of cytokine synthesis were evaluated by ELISA. UMWE showed an effect on increased splenocyte survival. UMWE elevated slightly PI3K phosphorylation and ERK1/2 phosphorylation used at 48 hr and 96 hr. Moreover, Bcl-2 was elevated at 48 hr and 96 hr in UMWE-treated splenocytes. UMWE decreased caspase-3 level at 48 hr and 96 hr. ICAD protein increased at 48 hr culturing time. Hematopoietin IL-2 cytokine was down-regulated, however IL-4 hematopoietin cytokine was up-regulated in UMWE treated cell culture media. Increased IFN-γ levels were verified in the supernatant of UMWE-treated cells in all periods (48 hr and 96 hr). Increased patterns in the production of IL-12 cytokine occurred as compared with control after 48 and 96 hr in UMWE-treated-cell cultures. These results suggested that UMWE can prolong splenocyte life span by controlling various signal factors and cytokines.

The effect of Injinchunggan-tang(Yinchenqinggan-tang) on Inflammatory Cytokine Gene Expression in Kupffer Cells (인진청간탕(茵蔯淸肝湯)이 kupffer cell 의 inflammatory cytokine 발현에 미치는 영향)

  • Kim Ji-Kwon;Kim Young-Chul;Lee Jang-Hoon;Woo Hong-Jung
    • The Journal of Internal Korean Medicine
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    • v.25 no.1
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    • pp.46-58
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    • 2004
  • Objectives : This study was designed to investigate the effects of Injinchunggan-tang(Yinchenqinggan-tang) on the expression of inflammatory cytokine genes and proteins in kupffer cells. Materials and Methods : The mRNA expression level and protein secretion level were measured using quantitative RT-PCR and ELISA assay respectively in Injinchunggan-tang-treated and untreated kupffer cells after exposed to ethanol, acetaldehyde and lipopolysaccharide. Results : Injinchunggan-tang(Yinchenqinggan-tang) reduced mRNA expression level and protein secretion level of $TNF-{\alpha},\;TGF-{\beta}1,\;IL-1{\beta},\;IL-6,\;IL-8$ that are induced by ethanol, acetaldehyde and lipopolysaccharide in kupffer cells and that mediate inflammation and fibrosis of liver. Conclusion : The result indicates that Injinchunggan-tang (Yinchenqinggan-tang) blocks alcohol-induced liver injury and protects liver by reducing production of inflammatory cytokines.

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The Effect of Injinchunggan-tang on Hepatitis C Virus Infection (인진청간탕이 C형 간염 바이러스의 임파구계 세포감염에 미치는 영향)

  • 김선민;이장훈;우홍정
    • The Journal of Korean Medicine
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    • v.22 no.1
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    • pp.53-62
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    • 2001
  • Objective : The aim of this study was to evaluate the efficacy of Injinchunggan-tang on Hepatitis C virus infection, and to clarify the mechanism of treatment by indentifying the effect of Injinchunggan-tang on cytokine secretion. Methods : In vitro model of HCV infection in MOLT 4 cell was used. The effect of Injinchunggan-tang on the attachment of HCV on MOLT 4 cell was studied by PCR method. The change of cytokine secretion according to Injinchunggan-tang treatment was investigated by ELISA. Results : Injinchunggan-tang inhibited the attachment of HCV on MOLT 4 in the concentration of $10-2{\mu\textrm{g}}/\mu\textrm{\ell}$ and $10-1{\mu\textrm{g}}/\mu\textrm{\ell}$. In cytokine assay, Injinchunggan-tang increased the secretion of IL-4 of mouse splenocytes and PBMC in 48 hour culture as well as the secretion of IL-12 of mouse splenocytes and PBMC in 48 hour culture, whereas it decreased the secretion of $IFN-{\gamma}$ of mouse splenocytes in 24 and 48 hour culture. Conclusion : The results of this study show that Injinchunggan-tang has an inhibitory effect on the attachment on HCV on Mo1t4 Cell, and that it increases the secretion of IL-4 and IL-12 of mouse splenocyte and PBMC.

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Effects of Water Extract Acorn on Mouse Immune Cell Activation Ex Vivo (도토리 추출물의 경구 투여가 마우스 면역 세포 활성에 미치는 효과)

  • Ryu, Hye-Sook
    • The Korean Journal of Food And Nutrition
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    • v.23 no.2
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    • pp.135-140
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    • 2010
  • Acorns have been used as a traditional remed as well as food source. However, few studies on their immunomodulating effects have been reported. In this study, the combined immunomodulative effect of a water extract of acorns was tested on seven to eight weeks old mice(balb/c). The mice were fed ad libitum on a chow diet, and a water extract of the plant mixture was orally administered every other day for four weeks at two different concentrations(50 and 500 mg/kg B.W.). The production of cytokine(IL-$1{\beta}$, IL-6, IL-2, IL-10, IFN-$\gamma$), secreted by macrophages stimulated with LPS or not, detected by ELISA assay using cytokine kit. After 48 h of incubation with mitogen(ConA or LPS) ex vivo study showed that cytokine (IL-$1{\beta}$, IL-6, IL-2, IL-10, IFN-$\gamma$) was detected in both of the 50 and 500 mg/kg B.W. supplementation groups with LPS stimulation. The results of this study may suggest that supplementation with acorn water extract increase immune function by regulating cytokine production capacity by activated macrophages.

Effect of Asterina pectinifera Lectin on Cytokine Production (별불가사리(Asterina pectinifera) 렉틴의 사이토카인 생성 양상)

  • 전경희;최수정;정시련
    • YAKHAK HOEJI
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    • v.43 no.4
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    • pp.474-480
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    • 1999
  • The purpose of this study is to define whether Asterina pectinifera Lectin (APL) is effective on the cytokine production. Isolated mRNA from hPBMC (human peripheral blood mononuclear cells) stimulated with APL for various reaction times (1 to 96 hours) was detected by RT-PCR. The intensity of band for IL-1 and $IFN{\gamma}$ mRNA was markedly increased at l hour, and IL-2 mRNA was strongly expressed at 4 hours. The mRNA band of APL-induced IL-2 and $IFN{\gamma}$ was weaker than that of IL-1, IL-6 and $TNF{\alpha}$. The mRNA expression of 4 cytokines (IL-1, IL-2, $IFN{\gamma}$ and $TNF{\alpha}$) was detected up to 48 hours, and that of IL-6 was detected until 72 hours. ELISA was used to look protein secretion of the cytokine gene with IL-1, IL-2 and TNF$\alpha$expressed strongly in RT-PCR. The highest protein secretion was at 4 hours with IL-1, at 8 hours with IL-2 and at 4 hours with $TNF{\alpha}$. These results suggest that APL can induce the production of some cytokines and the immune response from PBMC was done within the first few hours of stimulation with APL.

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Studies on the Regulatory Effect of Cytokine Production in Taumin Patients with Cerebral Infarction by Cheongsimyeonjatang (청심연자탕(淸心蓮子湯)이 태음인(太陰人) 뇌경색증(腦硬塞症) 환자(患者)의 세포활성물질(細胞活性物質) 생성조절(生成調節)에 미치는 영향(影響))

  • Kim, Kyung-Yo;Noh, Hyun-Soo
    • Journal of Sasang Constitutional Medicine
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    • v.12 no.2
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    • pp.162-170
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    • 2000
  • Purpose : Studies on the Regulatory Effect of Cytokine Production in Taumin Patients with Cerebral Infarction by Cheongsimyeonjatang Method : ELISA(enzyme-linked immuno-sorbent assay) Result : Chungsimyeunjatang(CYT) is a prescription for the cerebral infarction (CI) patients of Taeumin according to Sasang constitution philosophy. Taeumin patients with CI were treated with CYT during the acute stage. Clinical signs of CI disappeared markedly in about two to four weeks after oral administration of CYT in all patients. The mean interleukin (IL)-2 plasma levels were slightly lower in the patients with CI than in the normal groups, whereas the mean IL-4, IL-6 and IgE levels were significantly higher in the patients. There were no significant differences in $interferon-{\gamma}$ $(IFN-{\gamma})$ levels between the groups. Serum $IFN-{\gamma}$ and IL-2 levels derived from T helper (Th)1 cells were elevated significantly in the patients with CI by CYT administration. Significant reduced plasma levels of IL-4 and IL-6 derived from Th2 cells and IgE were observed in the patients treated with CYT. During the period of CYT administration, there were no other adverse effects. The data indicate that CYT has a good CI treatment effect, and that its action may be due to regulation of cytokine Production.

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Studies on the Regulatory Effect of Cytokine Production in Patients with Cerebral Infarction by Yangkyuksanhwatang (양격산화탕(凉膈散火湯)이 소양인(少陽人) 뇌경색증(腦硬塞症) 환자(患者)의 세포활성물질(細胞活性物質) 생성조절(生成調節)에 미치는 영향(影響))

  • Kim, Kyung-Yo;Kweon, Deog-Yun
    • Journal of Sasang Constitutional Medicine
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    • v.12 no.2
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    • pp.153-161
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    • 2000
  • 1. Purpose This studies the regulatory effect of cytokine production in Soyangin patients with cerebral infarction by Yangkyuksanhwatang. 2 Method ELISA 3. Result & Conclusion Yangkyuksanhwatang(YST) is a prescription for the cerebral infarction (CI) patients of Soyangin according to Sasang constitution philosophy. Soyangin patients with CI were treated with YST during the acute stage. Clinical signs of CI disappeared markedly in about 2 to 4 weeks after oral administration of YST in all patients. The mean interleukin (IL)-2 plasma levels were slightly lower in the patients with CI than in the normal groups, whereas the mean IL-4, IL-6 and IgE levels were significantly higher in the patients. There were no significant differences in interferon- ${\gamma}$ (IFN- ${\gamma}$ ) levels between the groups. Serum IFN- ${\gamma}$ and IL-2 levels derived from T helper (Th)1 cells were elevated significantly in the patients with CI by YST administration. Significant reduced plasma levels of IL-4 and IL-6 derived from Th2 cells and IgE were observed in the patients treated with YST. During the period of YST administration, there were no other adverse effects. The data indicate that YST has a good CI treatment effect, and that its action may be due to regulation of cytokine production.

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