Effects Of Angiopoietin-1 On Vascular Angiogenesis Regulator Factor In Human Umbilical Vein Endothelial Cells Cultured With High Glucose

Objective:To observe the expressional changes of angiopoietin-2 (Ang-2), pigment epithelium-derived factor (PEDF) and vascular endothelial growth factor (VEGF) in human umbilical vein endothelial cells (HUVECs) cultured with high glucose, and i nvestigate the effects of angiopoietin-1 (Ang-1) on these factors in high glucose.Methods:The HUVECs were divided into the following groups:normal group (5.5 mmol/L glucose), mannitol group (5.5 mmol/L glucose+24.5 mmol/L mannitol), high glucose group (30 mmol/L glucose) and Ang-1 group (30 mmol/L glucose+250 ng/ ml Ang-1). The expression of Ang-2, PEDF and VEGF were detected by immunofl uorescence, immunohisochemistry and real-time polymerase chain reaction (real-time PCR) at 24 and 48 hours after treated with different methods.Results:(1) Ang-2 was mainly expressed in the HUVECs cytoplasm and (or) membrane. The protein and mRNA of Ang-2 between the normal and mannitol group were not statistically different (P> 0.05); The levels of Ang-2 protein and mRNA in high glucose and Ang-1 groups were increased with time dependency, but significant difference were only found at 48h in high gloucose group (P<0.05); In the Ang-1 group, the levels of Ang-2 protein and mRNA at 48h were lower than that in high glucose group (P<0.05).(2) PEDF was mainly expressed in the HUVECs cytoplasm and (or) membrane. The protein and mRNA of PEDF between the normal and mannitol group were not statistically different (P>0.05); The levels of PEDF protein and mRNA in high glucose and Ang-1 groups were lower than that in the normal group (P<0.05), and the levels of PEDF in high glucose group were decreased with time dependency (48h vs 24h,.P<0.05); Compared to high glucose group, the increased levels of PEDF protein and mRNA in Ang-1 group were not statistically different (P> 0.05).(3) VEGF was mainly expressed in the HUVECs cytoplasm and (or) membrane. The protein and mRNA of VEGF between the normal and mannitol group were not statistically different (P>0.05); The levels of VEGF protein and mRNA in high glucose and Ang-1 groups were higher than that in the normal group (P<0.05), and the levels of VEGF in high glucose group were increased with time dependency (48h vs 24h, P<0.05); Compared to high glucose group, the decreased levels of VEGF protein and mRNA in Ang-1 group were not statistically different (P> 0.05).Conclusion:High glucose can induce abnormial expression of Ang-2, PEDF and VEGF in HUVECs in vitro, and may be with time dependency; Exogenous administration of Ang-1 can alleviate the abnormal expression of Ang-2, PEDF and VEGF in HUVECs cultured with high glucous.