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

Study on the antiangiogenesis effect of chondroitin sulfate isolated from Isostichopus badionotus

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SC-CHS对95D细胞增殖活性的影响;小管形成实验研究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)新生血管模型,研究其对血管生成的影响;通过RT-PCR和WesternBlot法检测其对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,这可能是通过影响肿瘤细胞中VEGF的合成发挥作用。

Objective To observe the antiangiogenesis effect of chondroitin sulfate（SC-CHS）isolated from Isostichopus badionotus,one of the bioactive compositions of sea cucumber body well,with various biological activities.Methods After being treated with different-concentrations of SC-CHS at different-time,the proliferation of 95D cells was assessed by MTT assay.Inhibition of angiogenesis was assessed by using tube-formation assays in SC-CHS treated HUVEC cells.CAM assays were used to assess inhibition effect of SC-CHS on physiological angiogenesis.The expressions 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,and could inhibit the tube formation as well as decrease the blood vessel growth on the chicken chorioallantoic membrane.SC-CHS could significantly down-regulate the expression levels of HIF-1α and VEGF mRNA when compared with control.The VEGF protein was also decreased.Conclusion SC-CHS exerted significant anti-angiogenic activities owing to their chemical structures and its mechanism is probably to inhibit the proliferation of 95D cells and reduce the expression of VGEF.