| 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7.5 mg/L。在斑马鱼(Danio rerio)胚胎体内实验中进一步表明, PCI在40 mg/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%。 |
| 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 |
| DOI： |
| 分类号: |
| 基金项目:国家自然科学基金资助项目( 30472043) |
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| Identification of a novel peptide from Coina intestinalis with potent antiangiogenetic activity |
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王 征,魏 宁
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| Abstract: |
| 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 Super dex 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 zebrafish embryos significantly in a dose-dependent manner and almost complete inhibition was observed when treated embryos with 40 mg/L PCI for 12h. |
| Key words: Coina intestinalis polypeptides antiangiogenesis zebrafish angiogensis model |
