Evaluation of the collaborative network of highly correlating skin proteins and its change following treatment with glucocorticoids.
BACKGROUND
Glucocorticoids (GC) represent the core treatment modality for many inflammatory diseases. Its mode of action is difficult to grasp, not least because it includes direct modulation of many components of the extracellular matrix as well as complex anti-inflammatory effects. Protein expression profile of skin proteins is being changed with topical application of GC, however, the knowledge about singular markers in this regard is only patchy and collaboration is ill defined.


METHODS
Scar formation was observed under different doses of GC, which were locally applied on the back skin of mice (1 to 3 weeks). After euthanasia we analyzed protein expression of collagen I and III (picrosirius) in scar tissue together with 16 additional protein markers, which are involved in wound healing, with immunhistochemistry. For assessing GC's effect on co-expression we compared our results with a model of random figures to estimate how many significant correlations should be expected by chance.


RESULTS
GC altered collagen and protein expression with distinct results in different areas of investigation. Most often we observed a reduced expression after application of low dose GC. In the scar infiltrate a multivariate analysis confirmed the significant impact of both GC concentrations. Calculation of Spearman's correlation coefficient similarly resulted in a significant impact of GC, and furthermore, offered the possibility to grasp the entire interactive profile in between all variables studied. The biological markers, which were connected by significant correlations could be arranged in a highly cross-linked network that involved most of the markers measured. A marker highly cross-linked with more than 3 significant correlations was indicated by a higher variation of all its correlations to the other variables, resulting in a standard deviation of > 0.2.


CONCLUSIONS
In addition to immunohistochemical analysis of single protein markers multivariate analysis of co-expressions by use of correlation coefficients reveals the complexity of biological relationships and identifies complex biological effects of GC on skin scarring. Depiction of collaborative clusters will help to understand functional pathways. The functional importance of highly cross-linked proteins will have to be proven in subsequent studies.
Klinge
Uwe
U
Surgical Department of the University Hospital of the RWTH Aachen, Germany. Uklinge@ukaachen.de
Farman
Nicolette
N
Fiebeler
Anette
A
D000818:Animals; D005260:Female; D005938:Glucocorticoids / Q000494:pharmacology*; D007150:Immunohistochemistry; D051379:Mice; D015999:Multivariate Analysis; D011506:Proteins / Q000378:metabolism*; D012867:Skin / Q000187:drug effects* / Q000378:metabolism
D005938:Glucocorticoids; D011506:Proteins