Title
In search of secreted protein biomarkers for the anti-inflammatory effect of β2-adrenergic receptor agonists: Application of DIGE technology in combination with multivariate and univariate data analysis tools
Author
Verhoeckx, K.C.M.
Gaspari, M.
Bijlsma, S.
van der Greef, J.
Witkamp, R.F.
Doornbos, R.P.
Rodenburg, R.J.T.
TNO Kwaliteit van Leven
Publication year
2005
Abstract
Two-dimensional difference gel electrophoresis (DIGE) in combination with univariate (Student's t-test) and multivariate data analysis, principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were used to study the anti-inflammatory effects of the β2-adrenergic receptor (β2-AR) agonist zilpaterol. U937 macrophages were exposed to the endotoxin lipopolysaccharide (LPS) to induce an inflammatory reaction, which was inhibited by the addition of zilpaterol (LZ). This inhibition was counteracted by addition of the β2-AR antagonist propranolol (LZP). The extracellular proteome of the U937 cells induced by the three treatments were examined by DIGE. PCA was used as an explorative tool to investigate the clustering of the proteome dataset. Using this tool, the dataset obtained from cells treated with LPS and LZP were separated from those obtained from LZ treated cells. PLS-DA, a multivariate data analysis tool that also takes correlations between protein spots and class assignment into account, correctly classified the different extracellular proteomes and showed that many proteins were differentially expressed between the proteome of inflamed cells (LPS and LZP) and cells in which the inflammatory response was inhibited (LZ). The Student's t-test revealed 8 potential protein biomarkers, each of which was expressed at a similar level in the LPS and LZP treated cells, but differently expressed in the LZ treated cells. Two of the identified proteins, macrophage inflammatory protein-1beta (MIP-1β) and macrophage inflammatory protein-1 alpha (MIP-1α) are known secreted proteins. The inhibition of MIP-1β by zilpaterol and the involvement of the β2-AR and cAMP were confirmed using a specific immunoassay. © 2005 American Chemical Society.
Subject
Pharmacology
Analytical research
β2-adrenergic receptor
Difference gel electrophoresis
Macrophage inflammatory protein-1β
Multivariate data analysis
Partial least squares discriminant analysis
Principal component analysis
Zilpaterol
adenylate kinase
beta 2 adrenergic receptor
beta 2 adrenergic receptor blocking agent
beta 2 adrenergic receptor stimulating agent
biological marker
butyryl cyclic AMP
clenbuterol
endotoxin
Escherichia coli lipopolysaccharide
formoterol
forskolin
heat shock protein
macrophage inflammatory protein 1alpha
macrophage inflammatory protein 1beta
phosphoglycerate mutase
propranolol
prostaglandin E2
proteome
salbutamol
unclassified drug
zilpaterol
adrenergic system
analytic method
antiinflammatory activity
article
controlled study
data analysis
discriminant analysis
human
human cell
inflammation
liquid chromatography
macrophage
mass spectrometry
multivariate analysis
nucleotide sequence
principal component analysis
priority journal
protein expression
protein secretion
proteomics
statistical analysis
two dimensional difference gel electrophoresis
two dimensional gel electrophoresis
Adrenergic Agonists
Adrenergic beta-Antagonists
Anti-Inflammatory Agents
Biological Markers
Cluster Analysis
Down-Regulation
Electrophoresis, Gel, Two-Dimensional
Enzyme-Linked Immunosorbent Assay
Humans
Immunoassay
Inflammation
Lipopolysaccharides
Macrophage Inflammatory Protein-1
Macrophages
Mass Spectrometry
Monocytes
Multivariate Analysis
Principal Component Analysis
Propranolol
Proteome
Proteomics
Receptors, Adrenergic, beta-2
Statistics
Trimethylsilyl Compounds
U937 Cells
Up-Regulation
Escherichia coli
To reference this document use:
http://resolver.tudelft.nl/uuid:37350a5a-20cb-45e1-9b38-fcdcff23d4c6
DOI
https://doi.org/10.1021/pr050183u
TNO identifier
238774
ISSN
1535-3893
Source
Journal of Proteome Research, 4 (6), 2015-2023
Document type
article