Overexpression of CREG Inhibits High Glucose Induced Apoptosis in Human Umbilical Vein Endothelial Cells

Objective: To investigate the role of CREG in high glucose induced human umbilical vein endothelial cells(HUVECs)injury, which may provide experimental evidence for exploring novel therapeutic targets for patients suffered from diabetic vasculopathy.Methods: Primary HUVECs were isolated by collagenase digestion and verified by CD31 immunofluorescence staining. HUVECs were treated with medium containing 5.5 mmol/l glucose(normal glucose control), 5.5 mmol/l glucose+27.5 mmol/l mannitol(osmotic control)and 33 mmol/l glucose(high glucose) for 48 h. Apoptosis of HUVECs was assessed by blotting cleaved caspase-3 or fluorescence activated flow cytometry(FACS) analysis of Annexin V/PI double immunostaining. Then HUVECs overexpressing CREG were established by infection with adenovirus carrying CREG gene. Effect of CREG overexpression on apoptosis of HUVECs was determined by detection of apoptosis. Results: Treatment with high glucose for 48 h resulted in elevated level of cleaved caspase-3 and higher apoptotic rate in HUVECs. After overexpression of CREG, cleaved caspase-3 and apoptosis rate in HUVECs treated with high glucose were significantly lowered, but still higher than those of normal glucose control group. Conclusion: Overexpression of CREG inhibits high glucose induced apoptosis in HUVECs.