体外三维抗血管生成药物筛选模型的建立

摘要/Abstract

摘要： 目的建立一种新的用于筛选具有抗血管生成作用的体外血管生成模型。方法人脐静脉的内皮细胞(HUVECs)在三维胶原纤维系统中培养,制备成具有毛细血管样网状结构模型;3种酪氨酸酶抑制剂genistein,tyrphostinA23和lavendustinC被分别加入到模型中,观察它们对血管形成的影响。结果在不加入任何生长因子的条件下,HUVECs能迅速重组形成毛细血管样网状结构;3种酪氨酸酶抑制剂均对血管形成有抑制作用。结论该模型具有操作简单、可重复性好、易量化的特点,是一种在体外筛选血管生成抑制剂的理想模型。

Abstract: Objective To develop a simple human angiogenesis model that simulates all the critical steps of the angiogenic process for screening antiangiogenic substances.Methods Human umbilical vein endothelial cells (HUVECs) were sandwiched into three-dimensional collagen gels for culturing to produce the angiogenesis model in vitro,on which the antiangiogenic effects of genistein,tyrphostin A23 and lavendustin C were assessed. Results HUVECs had the capacity to repidly form highly organized capillary-like structures in this model with uniformity of the entire culture,and was easily reproducible. Tyrosine kinase inhibitors,genistein,tyrphostin A23 and lavendustin C were found to inhibit the capillary tube formation.Conclusion This new human angiogenesis model has proved to be simple,responsive,and reproducible for testing potential angiogenic inhibitors,and is suitable for screening anti-angiogenic drugs.

中图分类号:

R733.7

李斌. 体外三维抗血管生成药物筛选模型的建立[J]. 南方医科大学学报, 2004, 24(10): 1167-1170.

LI Bin. Construction of a three-dimensional human angiogenesis model in vitro for antiangiogenic drug selection[J]. Journal of Southern Medical University, 2004, 24(10): 1167-1170.

参考文献

[1] Risau W.Mechanisms of angiogenesis[J].Nature,1997,386:671-4.
[2] Folkman J.Angiogenesis in cancer,vascular,rheumatoid and other disease[J].Nat Med,1995,(1):27-31.
[3] Folkman J,Haudenschild C.Angiogenesis in vitro[J].Nature,1980,288:551-6.
[4] Montesano R,Vassalli JD,Baird A,et al.Basic fibroblast growth factor induces angiogenesis in vitro [J].Proc Natl Acad Sci USA,1986,83:7297-301.
[5] Chalupowicz G,Chowdhury ZA,Bach TL,et al.Fibbrin Ⅱ induces endothelial cell capillary tube formation[J].J Cell Biol,1995,130:207-15.
[6] Kahn J,Mehraban F,Ingle G,et al.Gene expression profiling in an in vitro model of angiogenesis[J].Am J Pathol,2000,156:1887-900.
[7] Gimbrone MA,Cota RS,Folkman J.Human vascular endothelial cell in culture,growth and DNA synthesis[J].J Cell Biol,1974,60:673-84
[8] 吕家本,魏红军,蒋定文,等.用CAM技术评价鲨鱼软骨提取物抑制新生血管的活性[J].中国生化药物杂志,1998,19(4):173-5.Lu BJ,Wei HJ,Jiang DW,et al.Use chick embryo chorioallantoic membrane technique for evaluating inhitory activites of sark cartilage activated extract toword newangiogenesis [J].Chin J Biochem Pharmacol,1998,19(4):173-5.
[9] 黄煜伦,周幽心,周岱,等.雷公藤红素抑制血管生成的实验研究[J].中华肿瘤杂志,2003,25(5):429-31.Huang YL,Zhou YX,Zhou D,et al.Celatrol in the inhibition of neovasec ularization[J].Chin J Oncol,2003,25(5):429-31.
[10] 王玮琴,周慧君,陈欢欢,等.青蒿琥酯抑制新生血管生成的作用[J].中国药理学与毒理学杂志(Chin J Pharmacol Toxicol),2004,18(1):32-6.
[11] Grosios K,Holwell SE,McGown AT,et al.In vivo and in vitro evaluation of combretastatin A-4 and its sodium phosphate prodmg[J].Br J Cancer,1999,81 (8):1318-27.
[12] Ruggeri B,Singh J,Gingrich D,et al.CEP-7055:a novel,orally active pan inhibitor of VEGF receptor tyrosine kinase with potent antiangiogenic activity and antitumor efficacy in preclinical models[J].Cancer Res,2003,63 (18):5978-91.
[13] Friis T,Kjaer Sorensen B,Engel AM,et al.A quantitative ELISAbased co-cultured angiogenesis and cell proliferation assay [J].APMIS,2003,111:658-68.
[14] Montesano R,Orci L,Vassalli P.In vitro rapid organization of endothelial cells into capillary-like networks is promoted by collagen matrices[J].J Cell Biol,1983,97:1648-52.
[15] Cary LA,Chang JF,Guan JL.Stimulation of cell migration by overexpression of focal adhesion kinase and its association with Src and Fyn[J].J Cell Sci,1996,109(7):1787-94.