金丝桃素光动力通过抑制HUVEC细胞增殖,迁移和管腔形成抑制血管生成.docVIP

DOI：10.16016/j.1000-5404.201502036 金丝桃素光动力通过抑制HUVEC细胞增殖、迁移和管腔形成抑制血管生成 张 倩，李卓恒，李园园，李沉纹，管海燕，崔欢欢，李紫薇，卢来春 (400042 重庆，第三军医大学大坪医院野战外科研究所药剂科) ［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 cell, HUVEC）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HUVEC暗孵育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HUVEC细胞增殖的影响，Matrigel实验考察对HUVEC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HUVEC细胞增殖、迁移及管腔形成；HY-PDT使HUVEC中ERK1/2磷酸化比率(p-ERK1/2/ERK1/2)显著降低（P0.01），并显著下调Smad2 mRNA及蛋白表达（P0.01）。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HUVEC细胞增殖、迁移管腔形成受到抑制，抑制血管的生成。 ［关键词］ 金丝桃素；光动力；HUVEC；增殖；迁移；血管生成 [中图法分类号] [文献标志码] A Hypericin-photodynamic therapy inhibits angiogenesis through inhibiting HUVEC cell proliferation, migration and lumen forming Zhang Qian, Li Zhuoheng, Li Yuanyuan, Li Chenwen, Guan Haiyan, Cui Huanhuan, Li Ziwei, Lu Laichun (Department of Pharmacy, Institute of Surgery Research, Daping Hospital, Third Military Medical University, Chongqing, 400042, China) [Abstract］ Objective To investigate the effect of hypericin-photodynamic therapy (HY-PDT) on human umbilical vein endothelial cell (HUVEC) proliferation and migration as well as lumen forming and explore the mechanism of HY-PDT in anti-angiogenesis. Methods Hematoporphyrin (HP) was used as the positive control. HUVECs were incubated in media containing HY or HP for 24 h in the dark. The cells were then irradiated using a 585 nm light at a dose of 1.0 J/cm2. After the treatment, the cells were incubated for an additional 24 h in the dark. Cell viability was determined by MTT assay. To detect the effect of HY-PDT on lumen forming, Matrigel experiment was implemented. To test the mRNA expression of Smad2 and protein levels of ERK1/2, p-ERK1/2 and Smad2, quantitative real-time PCR (qRT-PCR) and Western blot were carried out, respectively. Results HY-PDT inhibited HUVEC cell p