血管生成调节蛋白-2在高糖诱导人脐静脉内皮细胞血管生成与转染中的表达和作用

Expression and role of angiogenesis regulatory protein-2 in angiogenesis and transfection induced by high glucose in human umbilical vein endothelial cells

目的研究血管生成调节蛋白-2(VASH2)在高浓度葡萄糖诱导人脐静脉内皮细胞(HUVECs)血管生成中的表达和作用。方法转染前,将HUVECs分为3组:正常对照组、高糖组、甘露醇组。将VASH2阴性对照质粒和siVASH2分别转染至高糖诱导的HUVECs,分别命名为si-NC组和si-VASH2组,未经处理的细胞为空白组。用实时荧光定量-聚合酶链反应法检测各组VASH2与血管生成相关因子基因表达; MTT法检测HUVECs增殖情况。结果转染前,正常对照组、甘露醇组和高糖组HUVECs中VASH2基因表达分别为1. 02±0. 34,1. 10±0. 37和1. 67±0. 49,高糖组与正常对照组和甘露醇组比较,差异均有统计学意义(均P <0. 05)。转染后,空白组、si-NC组和si-VASH2组VASH2基因表达水平分别为1. 71±0. 57,1. 68±0. 55和1. 01±0. 34;这3组的VEGFR2基因表达水平分别为1. 77±0. 58,1. 76±0. 57和1. 05±0. 35。在48h,空白组、si-NC组和si-VASH2组的HUVECs增殖率分别为(20. 34±6. 74)%,(20. 65±6. 75)%和(5. 53±1. 51)%。转染后的3项指标,si-VASH2组与空白组和si-NC组比较,差异均有统计学意义(均P <0. 05)。结论 VASH2在高糖诱导HUVECs中高度表达,干扰VASH2表达后可抑制血管生成相关因子表达,进而参与血管生成过程。

Objective To investigate the expression and effect of angiogenic regulatory protein-2(VASH2)in high glucose-induced angiogenesis of human umbilical vein endothelial cells(HUVECs).Methods HUVECs were divided into three groups:normal control group,high glucose group and mannitol group before transfection.The VASH2 negative control plasmid and si-VASH2 were transfected into high glucose-inducing HUVECs,respectively;named si-NC group and si-VASH2 group,and untreated cells were named blank group.The expression of VASH2 and angiogenesis-related factors in each group were detected by real time fluorescence quantitative polymerase chain reaction.Proliferation of HUVECs was detected by MTT assay.Results Before transfection,the expressions of VASH2 mRNA in HUVECs of normal control group,mannitol group,and high glucose group were 1.02±0.34,1.10±0.37 and 1.67±0.49,respectively;the differences between high glucose group with normal control group and mannitol group were statisticallysignificant(P<0.05).After transfection,the expressions of VASH2 in blank group,si-NC group and si-VASH2 group were 1.71±0.57,1.68±0.55 and 1.01±0.34,respectively;the expressions of VEGFR2 in the three groups were 1.77±0.58,1.76±0.57 and 1.05±0.35,respectively;the proliferation rates of HUVECs after transfection for48 h in the three groups were(20.34±6.74)%,(20.65±6.75)%and(5.53±1.51)%,respectively;the differences of the three factors between si-VASH2 group with blank group and si-NC group were statistically significant(all P<0.05).Conclusion VASH2 is highly expressed in HUVECs induced by high glucose,and interfering with the expression can inhibit the expressions of angiogenesis-related factors,thus participating in the angiogenesis process.