玻璃海鞘多肽的分离纯化及其抗血管生成活性  

Identification of a novel peptide from Coina intestinalis with potent antiangiogenetic activity

作　　者：董平原[1,2] 王征[1,2] 魏宁[1] 牛荣丽[1] 林秀坤[1] 

机构地区：[1]中国科学院海洋研究所,山东266071 [2]中国科学院研究生院,北京100039

出　　处：《海洋科学》2008年第8期48-52,共5页Marine Sciences

基　　金：国家自然科学基金资助项目(30472043)

摘　　要：利用丙酮沉淀，SephadexG25柱层析、Superdex75快速蛋白纯化系统（FPLC）等分离手段，采用活性追踪的方法，从玻璃海鞘（Coina intestinalis）中分离纯化出抗血管生成多肽PCI，据保留时间计算其分子质量为1．8ku。PCI在50～90℃范围内有良好的热稳定性，多肽水解酶处理可引起活性丧失。四甲基偶氮唑盐比色（MTT）检测表明其对人脐静脉血管内皮细胞（HUVEC）具有强烈的抑制作用，IC50为7．5mg／L。在斑马鱼（Danio rerio）胚胎体内实验中进一步表明，PCI在40mg／L的浓度下作用12h，斑马鱼胚胎新生血管生成受到显著抑制，肠下静脉血管长度为正常组的30％，斑马鱼胚胎血管生成率为正常组的45％。An antiangiogenesis peptide （PCI） was isolated from the tunicate, Coina intestinalis. The tunicate tissue was homogenized and extracted with 70% acetone solution following by gel chromatography of Sephadex G25. After FPLC purification on Superdex 75, a peptide with molecular weight 1.8 ku was obtained. Physiological study confirmed that the peptide is thermo-stable in the range of 50-90℃. In contrast, the activity of PCI could be destroyed by treatment with amino peptidase. Cell proliferation assay, using MTT method, showed that the concentrations of PCI required for 50% growth inhibition of HUVECs was 7.5 mg/L. In vivo experiment revealed that PCI inhibited the growth of SIV of zehrafish embryos significantly in a dose-dependent manner and almost complete inhibition was observed when treated embryos with 40 mg/L PCI for 12 h.

关 键 词：玻璃海鞘(Coina intestinalis) 多肽 抗血管生成 斑马鱼血管生成模型 

分 类 号：TQ02[化学工程]