## 4 Results
### HIVEC Treatment with Riaroxaban: Changes in the Expression of \(CYP4F2\)  
To determine the effect of rivaroxaban on HUVECs and to evaluate its association with vascular inflammation, cells were treated with 0.1, 0.2, 0.5, 1, 2, 5 and 10 \(\upmu\)M concentrations of rivaroxaban. Changes in \(CYP4F2\) and \(HNF4\alpha\) gene expressions were assessed in treated cells.  
\begin{table}
\begin{tabular}{c c c c c c c c} \hline \hline \multirow{3}{*}{**Metabolites**} & \multicolumn{6}{c}{**Retention Time (min)**} \\ \cline{2-9}  & **0.1 \(\upmu\)M** & **0.2 \(\upmu\)M** & **0.5 \(\upmu\)M** & **1 \(\upmu\)M** & **2 \(\upmu\)M** & **5 \(\upmu\)M** & **10 \(\upmu\)M** \\  & **Sample** & **Sample** & **Sample** & **Sample** & **Sample** & **Sample** & **Sample** \\ \hline M-1 & 0.33 & - & - & 0.35 & - & 0.3 & - \\ \hline M-2 & - & - & - & - & - & 0.53 & - \\ \hline M-5 & 4.29 & 4.24 & - & 4.29 & - & - & - \\ \hline M-8 & - & 2.95 & 2.97 & - & 2.98 & - & - \\ \hline M-10 & 0.55 & 0.55 & 0.53 & - & 0.59 & - & 0.57 \\ \hline M-11 & 3.07 & 3.01 & - & - & - & 3.01 & - \\ \hline M-18 & 2.81 & 2.8 & 2.85 & 3.11 & 2.9 & 2.9 & 2.86 \\ \hline \hline \end{tabular}
\end{table}
Table 3: Detection of rivaroxaban’s metabolites and its retention time in HUVECs using UPLC-QTOF-MS method.  
After treatment with rivaroxaban for 24 h, the expression of \(CYP4F2\) in HUVECs compared to control cells had decreased in a drug concentration-dependent manner (Table 4). The expression of \(HNF4a\) in HUVECs was not detected.  
\begin{table}
\begin{tabular}{c c c c c} \hline \hline \multirow{2}{*}{**Treated Samples**} & \multicolumn{3}{c}{**Changes in CYP4F2 \(\Delta\)Ct**} \\ \cline{2-5}  & **Median** & **Minimum–Maximum Value** & **Decreased Expression of CYP4F2 (FC \(\pm\) SD)** & \(p\) **Value** \\ \hline Control sample & 15 & 14–16 & - & - \\ \hline
0.1 \(\mu\)M rivaroxaban & 15 & 15–16 & 1.44-fold (0.69 \(\pm\) 0.14) & 0.046 \\ \hline
0.2 \(\mu\)M rivaroxaban & 15 & 15–16 & 1.71-fold (0.58 \(\pm\) 0.10) & 0.026 \\ \hline
0.5 \(\mu\)M rivaroxaban & 15 & 15–16 & 1.89-fold (0.53 \(\pm\) 0.17) & 0.028 \\ \hline
1 \(\mu\)M rivaroxaban & 16 & 15–17 & 1.93-fold (0.52 \(\pm\) 0.17) & 0.013 \\ \hline
2 \(\mu\)M rivaroxaban & 16 & 15–17 & 2.05-fold (0.49 \(\pm\) 0.15) & 0.004 \\ \hline
5 \(\mu\)M rivaroxaban & 16 & 15–17 & 2.4-fold (0.42 \(\pm\) 0.21) & 0.009 \\ \hline
10 \(\mu\)M rivaroxaban & 16 & 15–17 & 2.72-fold (0.37 \(\pm\) 0.21) & 0.009 \\ \hline \hline \end{tabular}
\end{table}
Table 4: Relative changes in the expression (\(\Delta\)Ct) of \(CYP4F2\) after exposure to 0.1, 0.2, 0.5, 1, 2, 5, and 10 \(\mu\)M rivaroxaban in HUVECs compared to control cells.  
Figure 1: Chromatogram of control and treated with different rivaroxaban concentration samples measured by LC-QTOF-MS system.  
Figure 2: Number of samples used in gene expression assay.  
HIVEC Treatment with hsa-miR-24-3p Minic: Changes in the Expression of CYP4F2 and CYP4F2 Concentration  
After forward transfection with the hsa-miR-24-3p mimic, the expression of _CYP4F2_ in HUVECs compared to negative controls was reduced (HUVECs treated with negative control miRNA) (Table 5). _HNF4a_ expression in HUVECs was not determined after transfection with the hsa-miR-24-3p mimic.  
A significantly lower concentration of CYP4F2 was detected in HUVECs transfected with the hsa-miR-24-3p mimic than in the negative control of HUVECs (Table 6).