Influence of Hesperidin and Diosmin as Active Metabolites of Bioflavonoids on Venous Wall Remodeling in Primary Cultures of Endothelial Cells In Vitro

Objective. To study the effect of active metabolites of bioflavonoids (hesperetin and diosmetin) on remodeling markers (von Willebrand factor (vWF), platelet-endothelial cell adhesion molecule-1 (CD31), fibronectin (FN), vimentin (VIM), plasminogen activator inhibitor type 1 (PAI-1)) in primary culture of endothelial cells. Material and methods. The study was conducted on primary cultures of human umbilical vein endothelial cells (HUVEC) in vitro. We selected concentrations of substances considering literature data, serum content of active metabolites after intake of the drugs containing diosmin and hesperidin, as well as in vitro tests. Results. Hesperetin at a concentration of 0.5 μM and diosmetin at a concentration of 1.5 μM did not affect the amount of VIM in HUVEC cell cultures. Diosmetin at a concentration of 15 μM decreased the marker by 45.6% (p=0.0006). The combination of hesperetin (5 μM) and diosmetin (15 μM) decreased VIM by 38.8% (p=0.0016). Diosmetin at a concentration of 150 μM and hesperetin at a concentration of 50 μM decreased VIM by 60.2% (p=0.0001) and 49.5% (p=0.0004), respectively. Hesperetin, diosmetin and their combination at concentrations of 0.5 and 1.5 μM reduced the amount of vWF by 56.3, 43.6 and 22.3%, respectively (p<0.0001). Diosmetin at a concentration of 15 μM and hesperetin at a concentration of 5 μM decreased vWF by 53.9% (p<0.0001) and 50.9% (p=0.0001), respectively. Combination of diosmetin (150 μM) and hesperetin (50 μM) decreased vWF by 37.2% (p=0.001). Hesperetin at concentrations of 0.5 and 5 μM, as well as diosmetin at a concentration of 1.5 μM did not affect the amount of CD31. Higher concentration of diosmetin (15 μM) decreased CD31 by 32.7% (p=0.002). Combination of diosmetin (150 μM) and hesperetin (50 μM) led to a more significant decrease in CD31 by 53.4% (p< 0.0001). Hesperetin and diosmetin did not affect PAI-1 expression at concentrations of 0.5 and 1.5 μM, respectively. Higher concentration of hesperetin (5 μM) and diosmetin (15 μM) decreased PAI-1 by 38.2% (p=0.0044) and 38.3% (p=0.0064), respectively. Combination of diosmetin (150 μM) and hesperetin (50 μM) resulted in a 40.7% decrease of PAI-1 (p=0.0032). Hesperetin and diosmetin at concentrations of 0.5 and 1.5 μM had no effect on FN. Higher concentration of diosmetin (15 μM) decreased FN by 36.5% (p=0.0004). Combination of diosmetin (150 μM) and hesperetin (50 μM) resulted in a more significant decrease of FN (by 45.6%, p=0.0001). Conclusion. Hesperetin and combination of hesperetin and diosmetin significantly affect remodeling markers in primary endothelial cell cultures. Combination of two active metabolites (hesperetin and diosmetin) has a more obvious effect. This indirectly indicates the feasibility of combination of two components (hesperidin and diosmin) to influence remodeling processes in vascular endothelium.