...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Osteoarthritis and cartilage >An in-vitro screening assay for the detection of inhibitors of proinflammatory cytokine synthesis: a useful tool for the development of new antiarthritic and disease modifying drugs.
【24h】

An in-vitro screening assay for the detection of inhibitors of proinflammatory cytokine synthesis: a useful tool for the development of new antiarthritic and disease modifying drugs.

机译：用于检测促炎细胞因子合成抑制剂的体外筛选测定：开发新的抗关节炎和疾病改良药物的有用工具。

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

OBJECTIVE: This work targets the development of a new tool to help develop new anticytokine drugs that prevent or reduce the progression of arthritic diseases. The specific aim of our study was to establish a fast and reliable in vitro screening assay of cytokine synthesis inhibitors (TNFalpha, IL-1beta) which shows better correlation with enzyme assays than previously reported in vitro assays. The test system should be able to detect p38-MAP kinase inhibitors. MATERIAL AND METHODS: Human peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll density gradient centrifugation from human EDTA-potassium whole blood. Cells were adjusted at 1 x 10(6) cells/ml. PBMCs were stimulated with lipopolysaccharide (LPS; E. coli serotype 026:B6: 1 microg/ml) in the presence of test compound (10(-5)-10(-8)M) for 4h at 37 degrees C in a 5% CO(2)-incubator. Induced TNFalpha and IL-1beta protein were measured by ELISA. RESULTS: The following are representative examples of inhibitors which effect cytokine synthesis. Corticoid Dexamethasone inhibits IL-1beta and TNFalpha synthesis at IC(50) of 38 nM and 25 nM, respectively. ERK1/ERK2 inhibitor U0126 effects cytokine synthesis at IC(50) of 0.34 microM for IL-1beta production and 0.26 microM for TNFalpha synthesis.p38-MAP kinase inhibitor SB 203580 inhibits IL-1beta- and TNF-alpha-synthesis (IC(50)sof 0.052 microM and 0.46 microM) in the same degree as p38-MAP kinase activity (IC(50): 0.34 microM). Same results could be shown for SB 210313, which had same efficacy on IL-1beta and TNFalpha biosynthesis (IC(50)'s: 1.88 microM and 1.01 microM) and on p38-MAP kinase (IC(50): 6.85 microM). Also for SB 202190 this correlation in inhibition of IL-1beta and TNFalpha synthesis (IC(50)'s: 0.055 microM and 1.01 microM) and p38-MAP kinase inhibition (IC(50): 0.088 microM) could be shown. CONCLUSION: This study shows the screening assay using PBMCs stimulated with LPS for IL-1beta and TNFalpha synthesis is a reliable test system for the quantification of the effectiveness of new drugs modulating IL-1beta and TNFalpha synthesis which is mainly mediated by p38-MAP Kinase. These assay allows fast detection of IL-1beta and TNFalpha synthesis inhibitors with different modes of action, including p38-MAP kinase inhibitors. The results obtained with our in-vitro screening assay show good correlation with results from enzyme assays.
机译：目的：这项工作的目标是开发一种新工具，以帮助开发预防或减少关节炎疾病进展的新抗细胞因子药物。我们研究的具体目的是建立一种快速，可靠的细胞因子合成抑制剂（TNFα，IL-1beta）的体外筛选测定法，该方法与酶测定法的相关性比以前报道的体外测定法更好。测试系统应能够检测p38-MAP激酶抑制剂。材料与方法：通过人EDTA-钾全血的Ficoll密度梯度离心分离人外周血单个核细胞（PBMC）。将细胞调整为1 x 10（6）细胞/ ml。在测试化合物（10（-5）-10（-8）M）存在下，用脂多糖（LPS;大肠杆菌血清型026：B6：1 microg / ml）刺激PBMC，在37摄氏度，5摄氏度下刺激4小时％CO（2）培养箱。通过ELISA测量诱导的TNFα和IL-1β蛋白。结果：以下是影响细胞因子合成的抑制剂的代表性实例。皮质类固醇地塞米松在IC（50）抑制IL-1beta和TNFalpha合成分别为38 nM和25 nM。 ERK1 / ERK2抑制剂U0126在IC（50）时产生的细胞因子合成对于IL-1beta的产生为0.34 microM，对于TNFalpha的合成为0.26 microM.p38-MAP激酶抑制剂SB 203580抑制IL-1beta和TNF-α的合成（IC（50 （0.052 microM和0.46 microM）的程度与p38-MAP激酶活性（IC（50）：0.34 microM）相同。 SB 210313的结果相同，对IL-1beta和TNFalpha生物合成（IC（50）：1.88 microM和1.01 microM）和p38-MAP激酶（IC（50）：6.85 microM）具有相同的功效。同样对于SB 202190，可以显示出在抑制IL-1beta和TNFα合成（IC（50）：0.055 microM和1.01 microM）和p38-MAP激酶抑制（IC（50）：0.088 microM）方面的相关性。结论：这项研究表明，用LPS刺激的PBMC对IL-1β和TNFα合成的筛选试验是一种可靠的测试系统，用于量化主要由p38-MAP激酶介导的调节IL-1β和TNFα合成的新药的有效性。 。这些测定法可以快速检测具有不同作用方式的IL-1beta和TNFalpha合成抑制剂，包括p38-MAP激酶抑制剂。用我们的体外筛选测定获得的结果与酶测定的结果显示出良好的相关性。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号