Investigation of antiatherogenic and antiinflammatory activity of betulinic acid applied prophylactically and therapeutically in human umbilical vein endothelial cells induced by tumour necrosis factor alpha

Local inflammation after endothelial damage is the beginning of atherosclerosis and inflammation has an important role not only at the beginning but at every stage of the disease. After activation of the endothelial cell, cytokines, chemokines and adhesion molecules are secreted. In studies conducted to reduce monocyte adhesion and cell damage with various phytochemicals, it has been shown that the levels of molecules such as intracellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and nuclear factor kappa B (NF-κB) may decrease. Induction of human umbilical vein endothelial cells (HUVECs) with TNF-α constitutes a good model of coronary artery disease. In this study, we aimed to investigate the anti-inflammatory and anti-atherosclerotic activity of betulinic acid, which has a high anti-inflammatory effect, at the biochemical level in the HUVEC cell line induced with tumour necrosis factor-alpha (TNF-α) and created inflammation and atherosclerosis model. In human umbilical vein endothelial cell culture in which HUVEC cell line was induced with TNF-α and inflammation and atherosclerosis model was created, LD50 concentration of betulinic acid was administered to each group 6 hours before, simultaneously with and 6 hours after 10 ng/mL TNF-α administration. Nitric oxide, TNF-α, interleukin 1-beta (IL-1β), IL-10, ICAM-1, VCAM-1 and NF-κB levels in endothelial cells were measured ELISAmetrically after 24 hours' incubation. Analyses were performed using SPSS for Windows v22.0 package programme. Nitric oxide, TNF-α, IL-1β, ICAM-1, VCAM-1 and NF-κB levels increased in the TNF-α treated group (p<0.05). A statistically significant decrease in these parameters was observed in all three groups treated with betulinic acid compared to the TNF-α-treated group (p<0.05). IL-10 level did not change with TNF-α treatment (p>0.05) but increased with TNF-α and betulinic acid treatment (p<0.05). TNF-α administration increased the levels of inflammatory parameters in HUVEC cells, indicating that an endothelial damage model was formed. The decrease in the levels of inflammatory parameters with TNF-α and betulinic acid administration suggests that betulinic acid may have therapeutic and prophylactic effects on inflammation and endothelial damage development.