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目的研究海洋小球藻多糖的制备工艺及其抑制肿瘤血管生成的作用。方法比较多种提取方法提取多糖,确定最适提取工艺,经乙醇沉淀,并结合DEAE-Cellulose 52柱色谱分离纯化;采用创伤愈合法测定小球藻多糖对肿瘤细胞培养液诱导的内皮细胞迁移的抑制作用。结果选择闪式萃取结合酶法最适宜提取小球藻多糖,其最佳提取条件为胰蛋白酶酶用量为1.5%、酶解pH值为7.5、酶解温度为40℃、酶解时间为2h,经柱色谱分离得到的小球藻多糖在浓度50、100μg/mL时可抑制肿瘤细胞培养液诱导的内皮细胞迁移。结论采用闪式萃取-酶解结合柱色谱可获得小球藻多糖,其具有一定的抑制肿瘤血管生成活性。
Objective To study the preparation of polysaccharide from Chlorella vulgaris and its inhibitory effect on tumor angiogenesis. Methods The polysaccharides were extracted by a variety of extraction methods, the optimal extraction process was determined and ethanol precipitation was carried out. The polysaccharides were separated and purified by DEAE-Cellulose 52 column chromatography. The wound healing assay was used to determine the effect of Chlorella polysaccharides on the endothelial cell migration induced by tumor cell culture medium Inhibition. Results The optimal extraction conditions for chlorella polysaccharides were as follows: 1.5% trypsin, pH 7.5, enzymolysis temperature 40 ℃, enzymatic hydrolysis time 2h, Chlorella polysaccharides isolated by column chromatography inhibited the migration of endothelial cells induced by tumor cell culture medium at concentrations of 50 and 100 μg / mL. Conclusion Chromobacterium chlorella polysaccharides can be obtained by flash extraction-enzymolysis combined with column chromatography, which has a certain inhibitory activity on tumor angiogenesis.