During inflammation, proteins and lipids act in a concerted fashion, calling for 
combined analyses. Fibroblasts are powerful mediators of chronic inflammation. 
However, little is known about eicosanoid formation by human fibroblasts. The 
aim of this study was to analyze the formation of the most relevant inflammation 
mediators including proteins and lipids in human fibroblasts upon inflammatory 
stimulation and subsequent treatment with dexamethasone, a powerful 
antiphlogistic drug. Label-free quantification was applied for proteome 
profiling, while an in-house established data-dependent analysis method based on 
high-resolution mass spectrometry was applied for eicosadomics. Furthermore, a 
set of 188 metabolites was determined by targeted analysis. The secretion of 40 
proteins including cytokines, proteases, and other inflammation agonists as well 
as 14 proinflammatory and nine anti-inflammatory eicosanoids was found 
significantly induced, while several acylcarnithins and sphingomyelins were 
found significantly downregulated upon inflammatory stimulation. Treatment with 
dexamethasone downregulated most cytokines and proteases, abrogated the 
formation of pro- but also anti-inflammatory eicosanoids, and restored normal 
levels of acylcarnithins but not of sphingomyelins. In addition, the chemokines 
CXCL1, CXCL5, CXCL6, and complement C3, known to contribute to chronic 
inflammation, were not counter-regulated by dexamethasone. Similar findings were 
obtained with human mesenchymal stem cells, and results were confirmed by 
targeted analysis with multiple reaction monitoring. Comparative proteome 
profiling regarding other cells demonstrated cell-type-specific synthesis of, 
among others, eicosanoid-forming enzymes as well as relevant transcription 
factors, allowing us to better understand cell-type-specific regulation of 
inflammation mediators and shedding new light on the role of fibroblasts in 
chronic inflammation.