Rosuvastatin Inhibits Interleukin (IL)-8 and IL-6 Production in Human Coronary Artery Endothelial Cells Stimulated With Aggregatibacter actinomycetemcomitans Serotype b

• Autores: Diego Fernando Gualtero Escobar, Sergio M. Viafara Garcia, Sandra Johanna Morantes, Diana M. Buitrago, Octavio A. Gonzalez, Gloria Inés Lafaurie Villamil
• Localización: Journal of periodontology, ISSN 0022-3492, Vol. 88, Nº. 2, 2017, págs. 225-235
• Idioma: inglés
• Texto completo no disponible (Saber más ...)
• Resumen

  • Background: Rosuvastatin exhibits anti-inflammatory effects and reduces periodontal diseases and atherosclerosis; however, its role in regulating periodontopathogen-induced endothelial proinflammatory responses remains unclear. The purpose of this study is to determine whether rosuvastatin can reduce the proinflammatory response induced by Aggregatibacter actinomycetemcomitans (Aa) in human coronary artery endothelial cells (HCAECs).

    Methods: HCAECs were stimulated with purified Aa serotype b lipopolysaccharide (LPS) (Aa-LPS), heat-killed (HK) bacteria (Aa-HK), or live bacteria. Expression of Toll-like receptors and cellular adhesion molecules were evaluated by fluorometric enzyme-linked immunosorbent assay. Endothelial cell activation was evaluated by quantifying nuclear factor (NF)-kappa B-p65 and cytokine expression levels by quantitative polymerase chain reaction and flow cytometry. Effect of rosuvastatin in expression of the atheroprotective factor Krüppel-like factor 2 (KLF2) and cytokines were also studied using similar approaches.

    Results: HCAECs showed increased interleukin (IL)-6, IL-8, intercellular adhesion molecule 1, and platelet endothelial cell adhesion molecule 1 expression when stimulated with Aa-LPS or Aa-HK. NF-κB-p65 activation was induced by all antigens. Aa-induced IL-6 and IL-8 production was inhibited by rosuvastatin, particularly at higher doses. Interestingly, reduced IL-6 and IL-8 levels were observed in HCAECs stimulated with Aa in the presence of higher concentrations of rosuvastatin. This anti-inflammatory effect correlated with a significant increase of rosuvastatin-induced KLF2.

    Conclusions: These results suggest Aa-induced proinflammatory endothelial responses are regulated by rosuvastatin in a mechanism that appears to involve KLF2 activation. Use of rosuvastatin to prevent cardiovascular disease may reduce risk of endothelial activation by bacterial antigens.

    Clinical and experimental evidence supports a critical role for inflammation in initiation and progression of atherosclerosis.1 Periodontal disease is a chronic oral inflammatory condition that has been proposed as a risk factor for atherosclerosis.2 Accordingly, periodontal treatment improves this proatherogenic condition.3 In addition to inflammation being a critical driver of atherosclerosis, growing evidence suggests infection could also play an important role.4,5 Presence of live bacteria or bacterial components, such as DNA of recognized periodontopathogens (e.g., Porphyromonas gingivalis [Pg] and Aggregatibacter actinomycetemcomitans [Aa]), has been demonstrated in atherosclerotic lesions.6,7 Increased levels of endotoxemia are related to cardiovascular disease.8 Similarly, increased levels of systemic inflammatory markers are associated with early atherosclerosis and periodontal disease.9 Two periodontal bacteria have been recognized as potential inducers of atherosclerotic lesions, namely Pg and Aa.10,11 Aa is a Gram-negative, non-motile, facultative anaerobic coccobacillus that can be acquired at an early age and is highly associated with clinical attachment loss and development of aggressive periodontitis.12 Nevertheless, this microorganism has also been isolated from patients with advanced chronic periodontitis,13 and available literature suggests that serotypes a, b, and c occur much more frequently among oral isolates than serotypes d, e, and f.14 Serotype b (including JP2 clone): 1) is more frequently associated with periodontitis in some populations and with various non-oral infections; 2) exhibits an increased capacity to trigger T helper (Th)1 and Th17-type cytokine production in immune cells; and 3) has lipopolysaccharide (LPS) immunogenicity that is greater than that of other serotypes.15 Translocation of Aa into circulating blood after periodontal treatment has also been shown, and this periodontopathogen has been detected in atherosclerotic plaques.6,16 Aa is a member of the group of fastidious Gram-negative organisms termed the HACEK group (Haemophilus spp., Aa, Cardiobacterium hominis, Eikenella corrodens, and Kingella spp.), which is involved in infective endocarditis and immunologic/vascular manifestations.17 Recently, more atherosclerotic lesions were shown in E-deficient spontaneously hyperlipidemic (Apoeshl) mice after infection with Aa and its LPS.18 Aa-LPS isolated from different strains increased expression of inflammatory cytokines interleukin (IL)-6 and IL-8 in human umbilical venous endothelial cells (HUVECs).19 However, the potential mechanisms by which Aa contributes to atherosclerosis remain unclear.

    Statins are very efficient, broadly used drugs that reduce the appearance of new events in patients with cardiovascular disease risk.20 The main protective effect of statins is related to their ability to block cholesterol synthesis by inhibiting 3-hydroxy-3-methylglutaryl coenzyme A. Statins have additional pleiotropic properties (i.e., anti-inflammatory effects) that may play a crucial role in controlling the atherosclerotic process.21 Rosuvastatin is a next-generation statin with significant anti-inflammatory effects and the ability to reduce the number of new coronary events in patients with increased serum levels of inflammatory markers without hyperlipidemia.22,23 Although the main mechanisms associated with the cardiovascular protective effects of rosuvastatin remain unclear to date, it has been suggested rosuvastatin suppresses inflammatory responses through a mechanism involving inhibition of c-Jun N-terminal kinases and nuclear factor (NF)-kappa B activation and reduced expression of intercellular adhesion molecule (ICAM)-1, monocyte chemoattractant protein-1, IL-8, IL-6, and cyclooxygenase-2. However, these effects were evaluated in HUVECs.24 Control of proinflammatory responses by statins may also be associated with reduction of atherosclerosis via inhibition of critical NF-κB-regulated endothelial genes, such as endothelial cell adhesion molecules (ECAMs), chemokines, and cytokines, which contribute to aberrant leukocyte adhesion to the endothelium during early atherogenic events.25 Statins have also shown atheroprotective effects via transcription factor Krüppel-like factor 2 (KLF2),26 which regulates endothelial thrombotic function and is downregulated by proinflammatory cytokines in HUVECs.27,28 Interestingly, growing evidence indicates statin therapy has local effects that significantly reduce the periodontal clinical index of inflammation and the levels of IL-1β, tumor necrosis factor (TNF)-alpha, and IL-6 in gingival crevicular fluid.29,30 Nevertheless, evidence of the potential effect of rosuvastatin in inhibiting periodontopathogen-induced endothelial activation is not available. The purpose of this study was to determine the effect of Aa on expression of proinflammatory molecules (i.e., adhesion molecules, Toll-like receptors [TLRs], and cytokines/chemokines) associated with the pathogenesis of atherosclerosis in human coronary artery endothelial cells (HCAECs) and to evaluate the potential of rosuvastatin for inhibiting Aa-induced proinflammatory responses.