马粪海胆生殖腺中多糖的纯化方法及其抗肿瘤作用研究

目的 优化马粪海胆生殖腺中多糖的纯化方法,并评价其体外对肿瘤细胞的生长抑制作用.方法 马粪海胆生殖腺经水提取、木瓜蛋白酶结合Sevage法除蛋白、透析后醇沉、干燥制得马粪海胆总多糖,总多糖应用Sephacryl S-300凝胶柱色谱分离纯化;采用苯酚-硫酸法测定多糖含量;通过噻唑蓝(MTT)法测试多糖的体外抗肿瘤作用.结果 马粪海胆生殖腺的水提液应用木瓜蛋白酶结合Sevage法除蛋白效果最佳,木瓜蛋白酶最佳用量为100∶6(海胆脱脂粉与酶质量比),制得总多糖的纯度达90%;总多糖及分离得到的2个多糖体外对人肝癌Bel7402细胞表现出生长抑制作用.结论 该纯化方法步骤简单、易操作,所得多糖理化性质好、易溶于水,适用于马粪海胆生殖腺多糖的纯化.

Research on the purification methods and their antitumor function of the polysaccharides from Hemicentrotus pulcherrimus gonad

Objective To optimize the purification methods of the polysaccharides from Hemicentrotus pulcherrimus gonad,and to evaluate their cytotoxic activity in vitro against human cancer cell lines.Methods The polysaccharides in H.pulcherrimus gonad were extracted with water,and the protein was removed by enzymic hydrolysis of papain and Sevage method,followed by dialyzed in flowing distilled water,precipitated in water and fully dried in progress.The total polysaccharide was separated by column chromatography on Sephacryl S-300 gel.The content of polysaccharides was measured with phenol-sulfuric acid method,and the antitumor effect was tested by MTT method.Results The optimum deproteinate effects were obtained by combination of enzymic hydrolysis of papain and Sevage method,and the optimized amount of papain was 100∶6 (the ration of the masses of defatted powder to papain),and the content of the purified total polysaccharide was 90%.The total polysaccharide and two separated polysaccharides showed anti-proliferative activity of Bel7402 cell lines.Conclusion The determined purification methods were applicable for the polysaccharides from H.pulcherrimus gonad for the simple and convenient operation,and that the purified polysaccharides displayed good physicochemical property and water-solubility.