In vitro evaluation of anti-inflammatory effect of rosuvastatin in endothelial cells stimulated by lipopolysaccharide Aggregatibacter actinomycetemcomitans.

Objectives: The purpose of this study was to determine the effect of purified LPS from A.a (Aa-LPS) on the expression of pro-inflammatory molecules (i.e., adhesion molecules, Toll-like receptors and cytokines/chemokines) associated with the pathogenesis of atherosclerosis in human coronary artery endothelial cells (HCAEC), as well as evaluating the potential of Rosuvastatin (RSV) for inhibiting the A.a-induced endothelial responses.
Methods: Monolayers of HCAECs were stimulated with LPS of periodontopathogens as A.a for at different times. Unstimulated HCAECs and HCAECs stimulated with Escherichia coli and P. gingivalis W83 LPS were used as controls. The expression levels of TLR2, TLR4, ICAM-1, VCAM-1 and PECAM-1 were evaluated by fluorometric cell ELISA, and cell activation was evaluated by measuring the expression of the cytokines IL-8 and IL-6 in supernatans using flow cytometry and by assessing p65 nuclear transcription NFκB activation by ELISA. The effect of RSV in Aa-LPS-induced pro-inflammatory responses was also studied using similar experimental approaches. Differences between treatments were analyzed by t test Student. A p-value ≤ 0.05 was considered statistically significant.
Results: After stimulation with A. a LPS the HCAEC increases expression of TLR2, ICAM-1 and PECAM-1, produces pro-inflammatory cytokines IL6, IL8 and induce NFκB activation by determining the p65 nuclear level. RSV at concentration of 400μM significantly reduces expression of PECAM-1 and decreases the production of IL8 and IL6 in HCAECs cells stimulated with A.a LPS.
Conclusions: These results suggest that Aa-LPS plays a role in pro-inflammatory endothelial responses that could be contributing to the atherosclerotic process, and the use of statins (i.e., RSV) could be reducing the likelihood for Aa-induced pro-atherosclerotic endothelial responses.