血管生成素1对高糖培养内皮细胞血管生成调控因子表达的影响研究

Effects of angiopoietin-1 on vascular angiogenesis regulatory factor of endothelial cells incubated with high glucose

目的观察高糖培养人脐静脉内皮细胞(HUVECs)血管生成调控因子表达变化,探讨血管生成素1(Ang-1)对血管生成调控因子的影响。方法将HUVECs随机分为正常组(NC)、甘露醇组(MT)、高糖组(HG)和Ang-1组。加入不同处理因素后培养24、48h,采用免疫组织化学法及RT-PCR检测血管生成素2(Ang-2)、色素上皮衍生因子(PEDF)、血管内皮生成因子(VEGF)蛋白及mRNA表达水平。结果与NC组比较,24h时HG组Ang-2、VEGF的蛋白及mRNA表达均上调[(26.99±1.69)vs(23.22±1.51),P>0.05;(37.32±1.39)vs (29.29±0.78),P<0.05;(165.00±32.08)vs(108.00±9.64),P<0.05;(185.33±14.44)vs(113.33±16.48),P<0.05],且48h上调明显,PEDF两时点蛋白及mRNA表达下调[(30.72±1.32)vs(37.81±1.43),(26.73±0.86)vs(37.43±1.48),P<0.05;(69.00±12.12)vs(129.33±19.74),(30.33±6.69)vs (122.67±20.88),P<0.05]。与同时点HG组比较,Ang-1组的Ang-2、VEGF蛋白及mRNA降低,PEDF蛋白及mRNA升高(P<0.05)。结论高糖培养HUVECs导致血管生成调控因子VEGF、Ang-2、PEDF表达异常。外源性给予Ang-1可缓解高糖导致血管生成调控因子的异常表达。

Objective To observe the expression of angiogenesis regulators in human umbilical vein endothelial cells（HUVECs）incubated with high glucose,and to investigate the influence of angiopoietin-1（Ang-1）on angiogenesis regulators. Methods HUVECs were randomly divided into four groups：normal control group（NC）,mannitol group（MT）,high glucose group（HG）and Ang-1 group（Ang-1）.The expression of Ang-2,PEDF and VEGF were detected by immunohistochemistry and real-time polymerase chain reaction（RT-PCR）at 24 and 48 hours after treated with different processing factors.Results Compared with NC group,the expression of Ang-2,VEGF protein and mRNA were up-regulated at 24 h[（26.99±1.69）vs（23.22±1.51）,P0.05;（37.32±1.39）vs（29.29±0.78）,P0.05;（165.00±32.08）vs（108.00±9.64）,P0.05;（185.33±14.44）vs（113.33±16.48）,P0.05],and this increase was obvious at 48 hin HG group.The PEDF protein and mRNA were down-regulated at 24 h and 48 h[（30.72±1.32）vs（37.81±1.43）,（26.73±0.86）vs（37.43±1.48）,P0.05;（69.00±12.12）vs（129.33±19.74）,（30.33±6.69）vs（122.67±20.88）,P0.05].Ang-2 and VEGF protein and mRNA were lower in Ang-1 group than in HG group at the same time point.PEDF protein and mRNA were higher in Ang-1 group than in HG group at the same time point（P0.05）. ConclusionHigh-glucose cultured HUVECs resulted in abnormal expression of angiogenesis regulators VEGF,Ang-2,and PEDF.Treatment with Ang-1 could alleviate the abnormal expression of angiogenic regulatory factors caused by high glucose.