定心方经Ang1-Tie2通路干预人脐静脉内皮细胞血管生成的实验研究  

Influence of Serum Containing Dingxin Recipe on Blood Vessel Formation Through Regulating Ang1-Tie2 Pathways in Human Umbilical Vascular Endothelial Cells

作　　者：赵丹丹[1] 苏志杰[1] 周凤华[1] 程赛博[1] 张宇[1] 张蕾[1] 徐煜凌 郭绍东[1] 贾钰华[1] 

机构地区：[1]南方医科大学中医药学院,广东广州510515

出　　处：《中药新药与临床药理》2017年第2期177-182,共6页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金(81373574);国家自然科学基金青年基金(81403339);广东省自然科学基金(2014A030313354);广东省自然科学基金博士启动项目(2014A030310150);广东省普通高校特色创新项目C1031897

摘　　要：目的探讨定心方(DXR)含药血清对氧化低密度脂蛋白(OX-LDL)诱导损伤的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)Angiopietin/Tie2(Ang1-Tie2)通路及血管形成的影响,研究DXR防治动脉粥样硬化(atherosclerosis,AS)的机制。方法体外培养人脐静脉内皮细胞,采用OX-LDL 100 mg·m L-1损伤人脐静脉内皮细胞24 h,分别观察DXR含药血清对其迁移及其小管形成的影响。采用免疫印迹法(Western Blot)检测血管内皮细胞生长因子(VEGF)和血管紧张素(Angiotensin1,Ang1)以及Ang1受体Tie2的表达。采用鸡胚尿囊膜实验观察(Chick embryo chorioallantoic membrane)DXR对体外血管生成的影响。结果经DXR作用24 h后,DXR高剂量组(DXR1,30%浓度)、中剂量组(DXR2,15%浓度)、低剂量组(DXR3,5%浓度)、空白组HUVEC细胞迁移数目与模型组相比,DXR1和DXR2组与Model组相比迁移细胞数量减少,差异具有统计学意义(P<0.05),DXR3、DXR2、DXR1、空白组鸡胚尿囊膜实验中形成的一级和二级血管数目与Model组一、二级血管数目相比,DXR3、DXR2、DXR1与Model组相比,一级血管数量明显降低(P<0.05),二级血管数量明显降低(P<0.05)。Western Blot实验显示,与Model组比较,受DXR干预的HUVEC细胞中的Ang1蛋白和Tie2蛋白表达量上调,VEGF蛋白表达量下降。受DXR干预的HUVEC细胞形成小管密集程度明显低于Model组。结论体外实验结果表明,DXR具有抑制血管生成的作用,这种效应与其可以调控HUVEC细胞内与血管生成密切相关的Ang1,Tie2,VEGF有关联。Objective To investigate the influence and possible anti-atherosclerosis mechanism of serum containing Dingxin Recipe（DXR）on the angiopietin 1/Tie2（Ang1-Tie2）pathway and blood vessel formation in human umbilical vascular endothelial cells（HUVECs）injured by oxidized low-density lipoprotein（OX-LDL）.Methods HUVECs were cultured in vitro and stimulated with OX-LDL（100 mg·m L（-1）） for 24 h,and then were treated separately with the serum containing different concentrations of DXR for another 24 h. The expression of vascular endothelial growth factor（VEGF）, angiopoietin 1（Ang1） and Ang1 receptor Tie2 was detected by Western blotting method. Chick embryo chorioallantoic membrane model was used for the analysis of blood vessel formation in vitro. Results After treatment with DXR for 24 h, the number of migrated cells in high-concentration DXR group（DXR1, 30 %）, middleconcentration DXR group（DXR2,15 %）,low-concentration DXR group（DXR3,5 %）and blank group was lessthan that of the model group,and the difference was significant between DXR1,DXR2 groups and model group（P〈0.05）. The number of the primary and secondary vessels in chick embryo chorioallantoic membrane model in DXR1,DXR2,DXR3 groups was less than that in the model group（P〈0.05）. Western blotting results showed that the serum contained DXR up-regulated the expression of Ang1 and Tie2,down-regulated the expression of VEGF. The area of tube formation in DXR groups was less than that of the model group. Conclusion DXR can inhibit the angiogenesis in vitro,and the mechanism may be related to the regulation of angiogenesis-associated factors of Ang1,Tie2 and VEGF in HUVECs.

关 键 词：定心方 动脉粥样硬化 ANG1 TIE2 VEGF 人脐静脉内皮细胞 血管生成 

分 类 号：R285.5[医药卫生—中药学]