过表达CREG抑制高糖诱导的人脐静脉内皮细胞凋亡

目的：探讨E1A激活基因阻遏子（Cellular repressor of ElA-stimulated genes，CREG）在高糖引起的人脐静脉内皮细胞（Human Umbilical Vein Endothelial Cells，HUVECs）损伤中的作用，为寻找糖尿病血管病变新的治疗靶点提供实验依据。方法：采用胶原酶消化法分离原代HUVECs，并用内皮细胞标志物CD31免疫荧光染色进行鉴定。分别用含有5．5mmol／1葡萄糖（正常糖对照组）、5．5mmol／1葡萄糖＋27．5mmol／1甘露醇（渗透压对照组）或33mmol／l葡萄糖（高糖组）的培养液培养HUVECs48h。WesternBlot检测剪切体caspase-3表达；AnnexinV／PI双染后流式细胞术检测细胞凋亡。通过感染表达CREG基因的腺病毒获得CREG过表达的HUvECs，WesternBlot及流式细胞术评价CREG过表达对HUVECs凋亡的影响。结果：高糖处理48h后，HUVECs内剪切体caspase-3的蛋白表达增加，细胞凋亡率增加；过表达CREG后，高糖处理的HUVECs内剪切体Caspase-3表达和凋亡细胞比例均明显降低，但仍高于正常糖对照组。结论：CREG过表达可抑制高糖引起的HUVECs凋亡。

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/1 glucose （normal glucose control）, 5.5 mmol/1 glucose＋27.5 mmol/1 mannitol （osmotic control） and 33 mmol/1 glucose （high glucose） for 48 h. Apoptosis of HUVECs was assessed by blotting cleaved caspase-3 or fluorescence acti- vated 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 detec- tion 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 signifi- cantly lowered, but still higher than those of normal glucose control group. Conclusion： Overexpression of CREG inhibits high glucose induced apoptosis in HUVECs.