3.3 Changes in the protein expression of Twist1 and Tie2 in
primary HUVECs under normal or hyperoxia conditions after using a Twist1
inhibitor
In the first step of this experiment, we used a final concentration of
3μmol/L of harmine, as determined by CCK8 data. Statistical analysis
showed that this concentration had no significant effect on the
viability of cells, but it was uncertain whether this concentration of
harmine could inhibit the expression of Twist1 protein and further
inhibit the expression of the Tie2 protein downstream. Therefore,
western blot was used to detect the changes in protein expression of
Twist1 and Tie2 after the use of harmine. As shown in Figure 3, compared
with the control group, the expression of Twist1 protein in the normoxia
+ harmine group was significantly reduced (p < 0.05) (Figure
3A, 3B). Compared with the hyperoxia group, the expression of Twist1
protein in the hyperoxia + harmine group also decreased significantly (p
< 0.05) (Figure 3A, 3B). Twist1 is the regulator of Tie2, and
the trend for change with the Tie2 protein was consistent with that of
Twist1. Under both normal and hyperoxia conditions, the expression of
Tie2 decreased significantly after the addition of harmine (p
< 0.01, p < 0.05, respectively) (Figure 3A, 3C).
These results proved that 3μmol/L of harmine inhibited the expression of
Twist1 and Tie2 proteins, thus achieving an efficient interventional
effect.