岩藻糖基化海参硫酸软骨素抑制肿瘤血管新生作用的研究.docVIP

 岩藻糖基化海参硫酸软骨素抑制肿瘤血管 新生作用的研究# 李辉，王静凤，杨玉红，徐雷雷，薛长湖** 5 10 15 （中国海洋大学食品科学与工程学院，青岛 266003） 摘要：目的：从美国肉参(Isostichopus badionotus)中分离纯化岩藻糖基化海参硫酸软骨素( sea cucumber chondroitin sulfate, SC-CHS )，体外研究其对肿瘤诱导血管生成的抑制作用。方法： 采用 MTT 法检测了 SC-CHS 对 95D 细胞增殖活性的影响；小管形成实验研究了 SC-CHS 对 脐静脉内皮细胞(HUVEC)小管形成能力的抑制作用；采用鸡胚尿囊膜(CAM)新生血管实验， 研究了其对 CAM 血管生成的影响；通过 RT-PCR 和 Western Blot 法检测了肿瘤诱导血管生 成相关因子缺氧诱导因子 (hypoxia inducible factor-1α , Hif-1α )和血管内皮生长因子 (vascular endothelial growth factor, VEGF) mRNA 和蛋白的表达。结果：结果显示 SC-CHS 能 显著抑制 95D 细胞的增殖，能抑制 HUVEC 的小管形成作用和 CAM 血管新生。RT-PCR 和 Western Blot 检测结果表明，高剂量 SC-CHS 能显著降低 95D 细胞中 Hif-1α和 VEGF mRNA 的表达，减少 VEGF 蛋白的合成。结论：SC-CHS 体外能显著抑制肿瘤血管生成，其机制通 过影响肿瘤细胞中 VEGF 的合成发挥作用。 关键词：食品营养学；岩藻糖基化海参硫酸软骨素；小管形成；鸡胚尿囊膜；Hif-1α；VEGF 中图分类号：Q95-33 20 Study on the antiangiogenesis effect of chondroitin sulfate isolated from Isostichopus badionotus Li Hui, Wang Jingfeng, Yang Yuhong, Xu Leilei, Xue Changhu (College of Food Science and Engineering in Ocean University of China, Qingdao 266003) 25 30 35 40 Abstract: Objective The Objective was to observe the antiangiogenesis effect of chondroitin sulfate（SC-CHS）isolated from Isostichopus badionotus, one of the bioactive compositions of sea cucumber body well, is proved to have various biological activities. Methods After treated with different-concentration of SC-CHS at different-time，the proliferation of 95D cells was assessed by MTT assay. Inhibition of angiogenesis was assessed in vitro using tube-formation assays in SC-CHS treated HUVEC cells. In vivo, CAM assays were used to assess inhibition effect of SC-CHS on physilogical angiogenesis. The expression of HIF-1α and VEGF mRNA were detected by RT-PCR, and VEGF protein was detected by Western Blot. Results SC-CHS could inhibit the proliferation of 95D cell. SC-CHS could inhibit the tube formation in vitro and decrease the blood vessel growth on the chicken chorioallantoic membrane in vivo. SC-CHS could significantly down-regulate the expression levels of HIF-1α and VEGF mRNA