• 제목/요약/키워드: Inhibition ELISA

검색결과 275건 처리시간 0.021초

십육미류기음(十六味流氣飮)이 FRTL-5 갑상선 세포의 DNA와 cAMP의 합성 및 MHC-class II의 발현에 미치는 영향 (Effects of Sipyukmiyukieum on DNA Synthesis, cAMP Synthesis and MHC-class II Expression of FRTL-5 Thyroid Cells)

  • 김병우;이재은
    • 대한한방내과학회지
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    • 제26권2호
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    • pp.398-408
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    • 2005
  • Objective: Graves' disease encompasses hyperthyroidism and diffused goiter associated with auto-antibodies to the thyroid stimulating hormone(TSH) receptors. In clinical environment, treatments of Graves' disease have many side effects such as recurrence and hypothyroidism. We've studied the effects of Sipyukmiyukieum on DNA synthesis, cAMP synthesis, and MHC-class II expression of FRTL-5 thyroid cells were studied. Methods: DNA synthesis was investigated by using BrdU staining and cAMP synthesis by ELISA kit, and expression of $interferon-{\gamma}$ activated MHC class II by Flow cytometer. Results: After introduction of Sipyukmiyukieum, significant inhibition of DNA synthesis. cAMP synthesis, and expression of $interferon-{\gamma}$ activated MHC class II of FRTL-5 thyroid cells was observed. Conclusions: Judging from these results, Sipyukmiyukieum has potential as a potent herbal treatment for inhibiting the enlargement of goiter, synthesis of abnormal thyroidal hormones, and autoimmuine responses of Graves' disease.

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HL-60 세포에서 $TNF-{\alpha}$에 의한 MCP-1 발현에 미치는 Betulinic Acid의 효과 (The Effect of Betulinic Acid on $TNF-{\alpha}-induced$ MCP-1 Expression in HL-60 Cells)

  • 김경찬;이추희
    • 약학회지
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    • 제52권1호
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    • pp.37-42
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    • 2008
  • Betulinic acid, a naturally occurring pentacyclic triterpenoid, is found in abundance in the outer bark of white birch (Betula alba). In this study, we investigated if betulinic acid affects cytokine expression from activated macrophage cells. ELISA result showed that stimulation of HL-60 cells with proinflammatory cytokine such as $TNF-{\alpha}$ resulted in MCP-1 release into culture medium. In addition, transcriptional upregulation of MCP-1 in response to $TNF-{\alpha}$ was observed by RT-PCR analysis. However, incubation of HL-60 cells with betulinic acid prior to $TNF-{\alpha}$ treatment abrogated MCP-1 expression in transcription and translational level. Consistent with a number of studies which reported requirement of ERK activation for $TNF-{\alpha}$ expression, Western blot analysis showed that $TNF-{\alpha}-induced$ ERK activation was suppressed by pretreatment of HL-60 cells with betulinic acid. Taken together, our data indicate that betulinic acid exerts its anti-inflammatory effect through inhibition of $TNF-{\alpha}-induced$ ERK activation which is required for the subsequent MCP-1 release.

황련의 쥐 대식세포로부터 LPS에 의해 유도되는 nitric oxide 및 $TNF-{\alpha}$의 생성억제효과 (Inhibitory effects of Coptidis Rhizoma on the LPS-induced production of nitric oxide and $TNF-{\alpha}$ in mouse macrophage cells)

  • 정효원;박용기
    • 대한본초학회지
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    • 제21권2호
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    • pp.165-173
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    • 2006
  • Objectives : Coptidis Rhizoma has been known traditional medicine with antimicrobial activities. We investigated inhibitory effects of Coptidis Rhizoma extract on lipopolysaccharide(LPS)-induced nitric oxide production from mouse macrophages. Methods : After Coptidis Rhizoma extract was pretreated in BV2, mouse brain macrophages and RAW264.7 mouse macrophages, cells were activated with LPS. To investigate cytotoxicity Coptidis Rhizoma extract, cell viability was measured by MTT assay. The production of nitric oxide(NO) and inducible nitric oxide synthase(iNOS) was determined in each culture supernatant and mRNA by Griess reaction and RT-PCR. The production of $TNF-{\alpha}$ from cells was measured by ELISA. Results : Coptidis Rhizoma extract significantly inhibited LPS-induced NO production in BV2 and RAW264.7 cells. Coptidis Rhizoma extract also greatly suppressed mRNA expression of iNOS in BV2 and RAW264.7 cells activated by LPS. Conclusion : These data suggests that Coptidis Rhizoma extract may have an anti-inflammatory effect through the inhibition of NO production.

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초피(椒皮)의 RAW264.7세포에서의 LPS에 의해 유도되는 nitric oxide 및 전염증사이토카인 생성억제효과 (Inhibitory effects of Zanthoxylum piperitum on the LPS-induced production of nitric oxide and proinflammatory cytokines in RAW264.7 cells)

  • 박용기
    • 대한본초학회지
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    • 제21권4호
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    • pp.69-76
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    • 2006
  • Objectives : The fresh young leaves and dried fruits of Zanthoxylum piperitum (Korean name: Chopi) are used as diuretics, stomachies, anthelmintic and for the treatments of disorders of the digestive organ in Asia. We investigated inhibitory effects of Zanthoxylum piperitum extract on lipopolysaccharide(LPS)-induced production of nitric oxide(NO) and pro-inflammatory cytokines including $TNF-{\alpha}$ and $IL-1{\beta}$ from RAW264.7 mouse macrophage cells. Methods : After methanol extract of Zanthoxylum Fructus (Zanthoxylum extract) was pretreated in RAW264.7 cells, the cells were stimulated with LPS. Cell toxicity of Zanthoxylum extract was assayed bv MTT assay. The production of NO from the cells was measured in culture medium by Griess reaction. The production of $TNF-{\alpha}$ and $IL-1 \;{\beta}$ from the cells was measured in culture medium by ELISA. Results : Zanthoxylum Fructus extract greatly inhibited the production of inflammatory mediators such as NO, $TNF-{\alpha}$ and $IL-1{\beta}$ from LPS-stimulated RAW264.7 cells. Conclusion : This result suggests that Zanthoxylum extract may have an anti-inflammatory effect through the inhibition of inflammatory mediators.

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Conformation of Group "a" Epitope in Hepatitis B Surface Antigen

  • Chun, Mun-Ho;Park, Won-Bong;Bok, Jin-Woo;Kim, Ha-Won;Choi, Eung-Chil;Kim, Byong-Kak
    • Archives of Pharmacal Research
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    • 제15권4호
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    • pp.347-355
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    • 1992
  • To elucidate structure of group "a" epitope, mouse antibodies that express idiotype monoclonal antibody and anti-idiotype monoclonal antibody against the group specific "a" determinant were purified by hydroxyapatite column. To obtain hepatitis B surface antigens (HBsAg). HBsAg positive blood was sequencially purified by ammonium sulfate precipitation, hydroxyapatite, sepharose 4B column chromatography and ultracentrifugation. The major protein (p25) and glycoprotein (gp30) of HBsAg were isolated by concanavalin-A-sepharose 4B. The ability of p25-gp30 among the HBsAg to inhibit the idiotype-anti-idiotype reaction was dependent on conformation, since reduced and alkylated p25-gp30 virtualy lost their inhibitory capacity when compared to native HBsAg. The data suggest that hepatitis B antigen is a conformational antigen critically dependent upon the disulfide bonds of p25-gp30.

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발효유제품에서 박테리오파지의 특성, 검출과 제어 (The Characteristics, Detection and Control of Bacteriophage in Fermented Dairy Products)

  • 안성일;리합 아조니;트란 티 탄 후옌;곽해수
    • 한국축산식품학회지
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    • 제29권1호
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    • pp.1-14
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    • 2009
  • This study was to review the classification, detection and control of bacteriophage in fermented dairy products. Bacteriophage has lytic and/or lysogenic life cycles. Epidemiologically speaking, detected major phages are c2, 936 and p335. Among them p335 has been the largest concern in dairy industry. Traditionally, various analytical technologies, such as spot, starter activity, indicator test, ATP measurement and conductimetric analysis, have been used for the phage detection. In recent years, advanced methods such as flow cytometric method, petrifilm, enzyme linked immunosorbent assay (ELISA) and multiflex PCR diagnostic kit have been deveoloped. The phage contamination has been controlled by using heat, high-pressure treatment, and the combinations of heat and pressure, and/or chemical. Also some starter cultures with phage-resistant character have been developed to minimize the concentration of phages in dairy product. Bacteriophage inhibition media such as calcium medium was also mentioned. To prevent the contamination of bacteriophage in dairy industry, further researches on the detection and control of phage, and phage resistant starters are necessary in the future.

Distinct Pro-Apoptotic Properties of Zhejiang Saffron against Human Lung Cancer Via a Caspase-8-9-3 Cascade

  • Liu, Dan-Dan;Ye, Yi-Lu;Zhang, Jing;Xu, Jia-Ni;Qian, Xiao-Dong;Zhang, Qi
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권15호
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    • pp.6075-6080
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    • 2014
  • Lung cancer is the leading cause of cancer-related death worldwide. Here we investigated the antitumor effect and mechanism of Zhejiang (Huzhou and Jiande) saffron against lung cancer cell lines, A549 and H446. Using high performance liquid chromatography (HPLC), the contents of crocin I and II were determined. In vitro, MTT assay and annexin-V FITC/PI staining showed cell proliferation activity and apoptosis to be changed in a dose- and time-dependent manner. The inhibition effect of Jiande saffron was the strongest. In vivo, when mice were orally administered saffron extracts at dose of 100mg/kg/d for 28 days, xenograft tumor size was reduced, and ELISA and Western blotting analysis of caspase-3, -8 and -9 exhibited stronger expression and activity than in the control. In summary, saffron from Zhejiang has significant antitumor effects in vitro and in vivo through caspase-8-caspase-9-caspase-3 mediated cell apoptosis. It thus appears to have more potential as a therapeutic agent.

수지상세포에 대한 Callophyllis japonica 추출물의 면역조절효과 (Immunomodulatory Effects of Callophyllis japonica Ethanol Extract on Dendritic Cells)

  • 김미형;주홍구
    • IMMUNE NETWORK
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    • 제7권2호
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    • pp.95-100
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    • 2007
  • Background: A red seaweed, Callophyllis japonica has been traditionally eaten in the oriental area. In a recent study, it has been demonstrated that the ethanol extract of C. japonica have antioxidant activity. However, there are few studies about the effects of C. japonica on the function of immune cells. We investigated the immunomodulatory effects of C. japonica on the function of dendritic cells, the potent antigen-presenting cells. Methods: Bone marrow-derived dendritic cells (DCs) were used and the viability was measured by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay and trypan blue exclusion test. Cytokine and nitric oxide (NO) levels were determined by using ELISA and Griess reagent, respectively. The expression levels of DC surface markers were measured by flow cytometric analysis. Results: C. japonica ethanol extract did not significantly affect the DCs viability and the IL-12 production from DCs, irrespective of the presence of lipopolysaccharide (LPS). In addition, it did not significantly change the expression of DC surface markers. However, C. japonica ethanol extract significantly inhibited the LPS-induced NO production and also increased the proliferation of allogeneic lymphocytes activated by DCs. Conclusion: Our data suggests that C. japonica ethanol extract enhances the proliferation of allogeneic lymphocytes activated by DCs which is associated with inhibition of NO production from DCs induced by LPS.

Glucosamine Inhibits Lipopolysaccharide-induced Inflammatory Responses in Human Periodontal Ligament Fibroblasts

  • Kim, Eun Dam;Park, Hyun-Jung;Baek, Jeong-Hwa
    • International Journal of Oral Biology
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    • 제39권4호
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    • pp.221-228
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    • 2014
  • Glucosamine is commonly taken by the elderly without prescription as a nutritional supplement to attenuate the progression or symptoms of osteoarthritis. Previous studies demonstrated that glucosamine shows anti-inflammatory effects in tissues such as blood vessels and the heart. However, there have been few reports about the effects of glucosamine on oral inflammatory diseases. Therefore, in this study, the effects of glucosamine on lipopolysaccharide (LPS)-induced inflammatory responses were investigated using human periodontal ligament fibroblasts (HPDLFs). HPDLFs were incubated in the presence and absence of glucosamine (10 mM) for 24 h, followed by treatment with E. coli LPS (100 ng/ml) or vehicle. Quantitative RT-PCR and ELISA results showed that LPS exposure significantly increased the levels of IL-6 and IL-8 mRNA and protein, while the effect was significantly suppressed by glucosamine treatment. Glucosamine did not attenuate, but slightly increased, the LPS-induced activation of mitogen activated kinases (ERK, p38, JNK). However, it suppressed the LPS-induced increase in the DNA binding affinity and transcriptional activity of NF-${\kappa}B$. These results suggest that glucosamine exerts anti-inflammatory effects on HPDLFs exposed to LPS via inhibition of NF-${\kappa}B$ activity, necessitating further studies using animal periodontitis models.

무기환(戊己丸)의 RAW 264.7 세포에 대한 항염증작용 연구 (Anti-inflammatory Effect of Mugi-hwan Water Extract in RAW 264.7 Cells)

  • 김일현;최종환;이세원;송용선
    • 한방재활의학과학회지
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    • 제23권3호
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    • pp.27-35
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    • 2013
  • Objectives The aim of this study was to investigate anti-inflammatory activity of Mugi-hwan (MGH) Water Extract. Methods Cells were treated with 2 ug/ml of LPS 1 hour prior to the addition of MGH. Cell viability was measured by MTS assay. The production of NO was determined by reacting cultured medium with Griess reagent. The expression of COX-2, iNOS and MAPKs was investigated by Western blot, RT-PCR. The content of level of cytokines ($PGE_2$, IL-6, in media from LPS-stimulated Raw 264.7 cells was analyed by ELISA kit. Results MGH inhibited the production of NO, $PGE_2$, IL-6 as well as the expressions of iNOS, COX-2 in the murine macrophage, RAW 264.7 cells. MGH also had suppression effects of LPS induced MAPKs activation. Conclusions These results suggest that MGH has an anti-inflammatory therapeutic potential, which may result from inhibition of MAPK phosphorylation, thereby decreasing the expression of pro-inflammatory genes.