摘要
目的探讨金丝桃素光动力(hypericin-photodynamic therapy,HY-PDT)对人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)增殖、迁移及管腔形成的影响,研究抑制血管生成的作用机制。方法以血卟啉为阳性药,用金丝桃素及血卟啉分别与HUVECs避光孵育24 h后,给予585 nm黄光照射(1.0 J/cm2),24 h后采用MTT法检测不同浓度HY对HUVECs细胞增殖的影响,细胞划痕实验观察HUVECs迁移情况,Matrigel实验观察对HUVECs细胞管腔形成的作用,qRT-PCR检测Smad2基因mRNA的表达,Western blot检测ERK1/2、p-ERK1/2、Smad2表达变化。结果 HY-PDT能够抑制HUVECs细胞增殖、迁移及管腔形成;HY-PDT使HUVECs中ERK1/2磷酸化比率(p-ERK1/2/ERK1/2)显著降低(P<0.01),并显著下调Smad2 mRNA及蛋白表达(P<0.01)。结论 HY-PDT通过抑制ERK1/2蛋白磷酸化活化、降低Smad2 mRNA表达及蛋白水平,使HUVECs细胞增殖、迁移管腔形成受到抑制,抑制血管的生成。
Objective To determine the effect of hypericin-photodynamic therapy （H^-PDT） on the proliferation, migration and lumen forming in human umbilical vein endothelial ceils （HUVECs）, and investigate the underlying mechanism of HY-PDT in anti-angiogenesis. Methods HUVECs were incubated in the culture medium containing HY or hematoporphyrin （HP, positive control） for 24 h in the dark, and then were irradiated by a 585 nm light at a dose of 1.0 J/cm2. After the treatment, the ceils were incubated for an additional 24 h in the dark. Cell viability was determined by MTT assay. Cell scratch test was used to determine the migration of HUVECs. Matrigel assay was implemented to detect the effect of HY-PDT on lumen forming. The mRNA expression of Smad2, and the protein levels of ERK1/2, p-ERK1/2 and Smad2 were detected by quantitative real-time PCR （qRT-PCR） and Western blotting, respectively. Results HY-PDT inhibited the proliferation, migration and lumen forming in HUVECs. The ratio of p-ERK1/2/ERK1/2 was significantly reduced in HUVECs following HY-PDT （ P 〈 0.01 ）. Additionally, HY-PDT remarkably decreased the mRNA and protein expression levels of Smad2 （ P 〈 0.01 ）. Collclusiorl HY-PDT inhibits the proliferation, migration and lumen forming in HUVECs by inhibiting activation of ERK1/2 phosphorylation and Smad2 expression at boththe transcriptional and translational levels, and thereby leads to inhibition of angiogenesis.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2015年第11期1125-1130,共6页
Journal of Third Military Medical University
基金
重庆市应用开发计划项目(CSTC2014yykf A110022)~~
关键词
金丝桃素
光动力
HUVECS
增殖
迁移
血管生成
hypericin
photodynamic therapy
human umbilical vein endothelial cells
proliferation
migration
angiogenesis
