高等学校化学学报 ›› 2003, Vol. 24 ›› Issue (9): 1588.

• 论文 • 上一篇    下一篇

17400鲨鱼软骨血管生成抑制因子的纯化及生物学活性研究

曾锋1, 杨伟兴1, 崔昆燕2, 刘威1, 张展霞1, 余健秀3, 庞义3   

  1. 1. 中山大学化学与化学工程学院;
    2. 中山大学测试中心;
    3. 中山大学生命科学学院, 广州 510275
  • 收稿日期:2002-11-20 出版日期:2003-09-24 发布日期:2003-09-24
  • 通讯作者: 曾锋(1964年出生),男,博士,副教授,从事生物分析化学及环境徽生物学等方面的研究.E-mail:ceszf@zsu,edu.cn E-mail:ceszf@zsu.edu.cn
  • 基金资助:

    广州市科学技术委员会科技攻关项目(批准号:98-J-002-01)资助

Studies on Purification and Bioactivity of 17400 Shark Cartilage Angiogenesis Inhibitor

ZENG Feng1, YANG Wei-Xing1, CUI Kun-Yan2, LIU Wei1, ZHANG Zhan-Xia1, YU Jian-Xiu3, PANG Yi3   

  1. 1. School of Chemistry and Chemical Engineering;
    2. Instrumental Analysis and Research Center;
    3. School of Life Science, Zhongshan University, Guangzhou 510275, China
  • Received:2002-11-20 Online:2003-09-24 Published:2003-09-24

摘要: 采用盐酸胍抽提、膜超滤、丙酮分级沉淀、Sephadex-75柱层析和C4反相高效液相色谱等分离步骤,从广东阳江鲨鱼软骨中纯化获得新的鲨鱼软骨血管生成抑制因子SCAI-c(SharkCartilageAngiogenesis In-hibitor-c,SCAI-c);SDS-PAGE电泳银染显示为一条带,根据蛋白质的相对迁移率计算,分子量为17400;它对鸡胚绒毛尿囊膜血管的形成具有显著抑制效应,并有明显的浓度依赖关系.SCAI-c与SCAI-a具有类似的生物学特征.

关键词: 鲨鱼软骨, 血管生成抑制因子, 分离纯化, 生物活性, 鸡胚绒毛尿囊膜

Abstract: SCAI-c, another angiogenesis inhibitor from shark cartilage of Guangdong Yangjiang, was purified and the bioactivity of SCAI-c were investigated. The 1. 0 mol/Lguanidine and 0. 02 mol/L MES(at 28 ℃) extracts of shark cartilage was centrifuged at 6000 r/min for 30 min and the pellet was discarded, and the supernatant was recentrifuged at 12000 r/min for 30 min, then ultrafiltrated and fractionally precipitated with 25%-90% acetone. The precipitate was applied to a Sephadex G-75 gel column and eluted after the predominand peak of A280nmabsorbing material with 0. 05 mol/L Tris-HCl buffer (containing 0.02 mol/L NaCl and 0. 02% NaN3). The material Bwere composed of three single peptides with molecular mass of 17600, 15000, 12600, then were purified by using reverse-phase HPLC, the eluate of Cwas collected, dried and observed by chicken chorioallantoic membrane. The pure inhibitor was homogeneous as a single band on a silver-stained 12. 5% sodium dodecyl sulfate-polyacrylamide gel. SCAI-c has a molecular weight of 17600, and could specifically inhibit angiogenesis of the chorioallantoic membrane of chick embryos, and the inhibitory effect of angiogenesis was SCAI-c concentration dependent, but the concentration of SCAI-c increased to 0. 5 μg/μL, chick embryos died about to 20%-30%. Inhibitory characteristic of SCAI-c was similar with that of SCAI-a. They are specific angiogenesis inhibitor, and may be a useful drug on tumor therapy in some day.

Key words: Shark cartilage, Angiogenesis inhibitor, Separarion and purification, Bioactivity, Chorioallan-toic membrane of chick embryos

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