Background/Purpose: In certain chronic
diseases, such as rheumatoid arthritis (RA), atherosclerosis is more prevalent.
RA patients have a higher incidence of cardiovascular disease and at least two-fold
enhanced cardiovascular risk, as compared to the general population. This is in
part due to persistently present inflammation shown to be a risk factor for
atherosclerosis. Serum amyloid A (SAA) is an acute phase protein, highly
upregulated in sera of RA patients and reported to activate endothelial cells. Our
aim was to determine the effects of medications used in RA patients on selected
adhesion molecules, cytokines and chemokines involved in atherosclerosis and coagulation
in human coronary artery endothelial cells (HCAEC) following stimulation with SAA.
Methods: Primary HCAEC at passage 5, grown to confluency in 6-well
plates were preincubated for 30 min with medications from sterile ampules at
indicated final concentrations (Dexamethasone 1μM, Methotrexate 1μM, Certolizumab
pegol 100μg/ml, Etanercept 100μg/ml, Meloxicam 100μM, Diclofenac
10μM) or dissolved in medium (Captopril 10μM) or DMSO (Fluvastatin
10μM, Etoricoxib 100μM, Rosiglitazone 30μM). Human recombinant
SAA1/2 (Peprotech, London, UK) was used to stimulate HCAEC at a final 1000 nM concentration.
After 24 hours, supernatants were collected, centrifuged and frozen at -20 C.
IL-6, IL-8, sVCAM-1, PAI-I were measured by ELISA. The number of viable cells
was determined by colorimetry (CellTiter MTS assay, Promega). Results: SAA-stimulated levels of released
IL-6, IL-8 and VCAM-1 in HCAEC supernatants were significantly attenuated by Methotrexate,
Fluvastatin and Etoricoxib. Both Certolizumab pegol and Etanercept significantly
decreased PAI-1 by an average of 43%. Captopril did not significantly alter either
the cytokine or chemokine profile released by SAA stimulation of HCAEC. Rosiglitazone
caused only slight lowering of IL-6, but significantly inhibited VCAM-1 by 58%.
Among the NSAID group of medications, Diclofenac and Etoricoxib showed a trend
towards lowering IL-6 and IL-8, while Meloxicam showed just the opposite.
Diclofenac and Etoricoxib significantly attenuated VCAM-1 released supernatant
levels, while Meloxicam was less effective. Diclofenac and Etoricoxib increased
PAI-1, which was significantly reduced by Meloxicam. Conclusion:
The most successful agent in lowering cytokine
production was Fluvastatin. None of the medications significantly influenced cell
viability. Methotrexate also showed strong beneficial effects on lowering released
cytokine/chemokine/adhesion molecule levels from SAA-treated HCAEC. An interesting
duality was observed in the NSAID group of medications, with Meloxicam exhibiting
opposite effects from Diclofenac and Etoricoxib.   

Table 1: Influence of
medications used in RA patients on released IL-6, IL-8, sVCAM-1, PAI-1 protein
levels and viability of SAA-activated HCAEC compared with the influence of SAA
alone