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
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