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Related Experiment Videos

High throughput screening for intercellular adhesion molecule-1 inhibitor.

De-cheng Ren1, Guan-hua Du, Jun-tian Zhang

  • 1Institute of Materia Medica, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100050, China.

Yao Xue Xue Bao = Acta Pharmaceutica Sinica
|September 30, 2003
PubMed
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A new assay effectively identifies inhibitors of intercellular adhesion molecule-1 (ICAM-1) expression in human umbilical vein endothelial cells (HUVEC). This method distinguishes active compounds from cytotoxic effects, aiding drug discovery.

Area of Science:

  • Cell Biology
  • Immunology
  • Drug Discovery

Background:

  • Intercellular Adhesion Molecule-1 (ICAM-1) plays a crucial role in endothelial cell function and inflammatory responses.
  • Identifying modulators of ICAM-1 expression is vital for developing targeted therapies for inflammatory diseases.
  • High-throughput screening (HTS) assays are essential for efficiently evaluating large compound libraries.

Purpose of the Study:

  • To establish a robust, high-throughput screening assay for identifying inhibitors of ICAM-1 expression.
  • To validate the assay's performance using human umbilical vein endothelial cells (HUVEC).
  • To simultaneously assess compound efficacy and cytotoxicity.

Main Methods:

  • ICAM-1 expression was quantified using Enzyme-Linked Immunosorbent Assay (ELISA) in lipopolysaccharide (LPS)-stimulated HUVEC.

Related Experiment Videos

  • Compound cytotoxicity was evaluated using the MTT assay.
  • A library of 2,000 compounds was screened using the developed assay.

    • Main Results:

    • LPS stimulation significantly increased ICAM-1 expression in HUVEC in a dose- and time-dependent manner.
    • The HTS assay identified a hit rate of 1.5% among the 2,000 screened compounds.
    • Out of 30 identified hits, 24 compounds exhibited cytotoxic effects.

    Conclusions:

    • The developed ELISA-based assay is cost-effective, reproducible, and suitable for HTS in primary endothelial cells.
    • This assay effectively identifies ICAM-1 inhibitors while discriminating against cytotoxic compounds.
    • The assay provides a feasible platform for discovering novel therapeutic agents targeting ICAM-1.