Aim
To study the effects of LY294002 [phosphatidylinositol 3-kinase (PI3K) inhibitor] on the function and mechanisms of retinal endothelial cells (RECs) in vitro.
Conclusion
LY294002 regulates the function of RECs by reducing the expression of MMP-2, MMP-9, and VEGF in vitro. LY294002 may provide an effective method for preventing pathological angiogenesis.
Methods
RECs were randomly divided into control group and LY294002 treatment group. RECs in the control group were placed the incubator for hypoxic exposure in vitro. RECs in the LY294002 treatment group were pretreated with LY294002 (40 µmol/L) under hypoxic condition. The expression of matrix metalloproteinase (MMP)-2, MMP-9, vascular endothelial growth factor (VEGF), and apoptosis and proliferation of RECs were evaluated with Western blot, real-time reverse transcription-polymerase chain reaction (RT-PCR), and flow cytometric analysis, correspondently.
Results
Compared with the control group, treating the RECs with LY294002 was able to remarkably inhibit cell proliferation rates (t1d=2.13, t2d=2.65, t3d=2.36, t4d=2.06, all P<0.05). Flow cytometric analysis indicated that a moderate increase in apoptosis in the LY294002 treatment group compared to the control group (t=2.51, P<0.05). The expression of MMP-2, MMP-9 and VEGF were downregulated in the LY294002 treatment group by Western blot and real-time RT-PCR (all P<0.05).