A Combined Impedance and AlphaLISA-Based Approach to Identify Anti-inflammatory and Barrier-Protective Compounds in Human Endothelium
Autor: | Rudolf Lucas, Elisabeth Hofmann, Nico Jacobi, Jiri Kopecky, Anita Koppensteiner, Michael Katzlinger, Maren Pflüger, Aleksandra Kapuscik, Wolfgang Schütt, Kamil Önder, Christoph Wiesner, Harald Hundsberger, Andreas Eger, Jouni Jokela |
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Rok vydání: | 2013 |
Předmět: |
Transcriptional Activation
Chemokine Endothelium Pyridines Anti-Inflammatory Agents Gene Expression Inflammation Biology Cyanobacteria Peptides Cyclic Biochemistry Analytical Chemistry Microcirculation Capillary Permeability 03 medical and health sciences 0302 clinical medicine Depsipeptides Electric Impedance medicine Humans Interleukin 8 Lung 030304 developmental biology 0303 health sciences ICAM-1 Tumor Necrosis Factor-alpha Interleukin-8 Imidazoles Endothelial Cells Reproducibility of Results Intercellular Adhesion Molecule-1 High-Throughput Screening Assays 3. Good health Cell biology Endothelial stem cell Kinetics medicine.anatomical_structure 030220 oncology & carcinogenesis Microvessels biology.protein Cytokines Molecular Medicine Tumor necrosis factor alpha Endothelium Vascular medicine.symptom Cell Adhesion Molecules Biotechnology |
Zdroj: | SLAS Discovery. 18:67-74 |
ISSN: | 2472-5552 |
Popis: | Chronic inflammation is at least partially mediated by the chemokine-mediated attraction and by the adhesion molecule-directed binding of leukocytes to the activated endothelium. Therefore, it is therapeutically important to identify anti-inflammatory compounds able to control the interaction between leukocytes and the endothelial compartments of the micro- and macrocirculation. When testing novel drug candidates, it is, however, of the utmost importance to detect side effects, such as potential cytotoxic and barrier-disruptive activities. Indeed, minor changes in the endothelial monolayer integrity may increase the permeability of small blood vessels and capillaries, which, in extreme cases, can lead to edema development. Here, we describe the development of a high-throughput screening (HTS) platform, based on AlphaLISA technology, able to identify anti-inflammatory nontoxic natural or synthetic compounds capable of reducing tumor necrosis factor (TNF)-induced chemokine (interleukin [IL]-8) and adhesion molecule (ICAM-1) expression in human lung microvascular endothelial cells. Quantification of cell membrane-expressed ICAM-1 and of cell culture supernatant-associated levels of IL-8 was analyzed in HTS. In parallel, we monitored monolayer integrity and endothelial cell viability using the electrical cell substrate impedance sensing method. This platform allowed us to identify natural secondary metabolites from cyanobacteria, capable of reducing ICAM-1 and IL-8 levels in TNF-activated human microvascular endothelial cells in the absence of endothelial monolayer barrier disruption. |
Databáze: | OpenAIRE |
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