2019 Volume 18 Issue 1 Pages 92-100
Periodontitis has been associated with an increased risk for atherosclerosis.
Accumulating evidence suggests that endothelical dysfunction is an early marker for atherosclerosis. To determine how periodontal infection could contribute to endothelial dysfunction, we examined the ability of the major periodontal pathogen, Aggregatibacter actinomycetemcomitans (Aa) -sensitized monocytes to modulate properties of human umblical vein endothelial cells(HUVEC) by assessment of reactive oxygen species(ROS) production and apoptosis.
Cell proliferation and apoptosis was measured by BrdU cell proliferation ELISA kit, cell death detection ELISA and caspase activities. Detection of intracellular ROS generation
was evaluated flurometrically using H2DCF-DA. Quantitative reverse transcription polymerase chain reaction was performed using primers specific for p22phox and p47phox.
iNOS, p22phox and p47phox in cell lysates were detected by Western blot analysis with the respective specific antibodies.
Aa did not affect the viability of HUVEC but induced apoptosis in HUVEC cocultured with monocytes. Aa significantly induced ROS and NO productions in monocytes.
Furthermore, Aa increased gene and protein expressions of p22phox, p47phoxand iNOS.
Also, H2 O2 induced growth inhibition, apoptosis and caspase 3/7 activities in HUVEC.
These results suggest that apoptosis in HUVEC could be induced by Aa -sensitized monocytes via ROS-dependent pathway, potentially amplifying proatherogenic mechanism in the perturbed vasculature.
