龙葵多糖细胞毒活性物质基础研究

Material basis of cytotoxicity in polysaccharide from Solanum nigrum

目的研究龙葵多糖细胞毒活性的物质基础。方法从龙葵青果中分离出龙葵粗多糖;Sevage法除去游离蛋白;10%H2O2脱色,95%乙醇沉淀,分离出龙葵多糖。龙葵多糖经DEAE-52纤维素柱色谱分离得多糖-蛋白复合物;采用SDS-PAGE电泳法检测多糖-蛋白复合物是否为糖蛋白,并测定其相对分子质量;MTT法检测多糖-蛋白复合物对乳腺癌细胞MCF-7的IC50;多糖-蛋白复合物再经SephadexG-200凝胶柱色谱精制,采用MTT法进行活性检测。结果 SDS-PAGE电泳法测得龙葵多糖-蛋白复合物是相对分子质量3.0×104和2.5×104的2种糖蛋白的复合物,经MTT法测得其对乳腺癌细胞MCF-7的IC50为804.51μg/mL;多糖-蛋白复合物经SephadexG-200凝胶柱色谱精制得糖蛋白A、B,并测得其对MCF-7的IC50分别为532.96、613.91μg/mL。结论龙葵多糖细胞毒活性的物质基础是相对分子质量为3.0×104和2.5×104的2种糖蛋白。

Objective To study the material basis of cytotoxicity in polysaccharide from Solanum nigrum. Methods Crudeproducts of polysaccharide were isolated from S. nigrum fruit with free protein being removed by Sevage method. The crude products were decolored by 10% H2O2 and precipitated by 95% ethanol treatment. Polysaccharide-protein complex was isolated by DEAE-52 fiber column and the relative molecular weight was detected by SDS-PAGE. MTT assay was used in detecting the cytotoxicity of polysaccharide-protein complex for MCF-7 cells in vitro. Then polysaccharide-protein complex was separated by SephadexG-200 gel column and the cytotoxicity was detected by MTT method. Results Polysaccharide-protein complex was isolated by DEAE-52 fiber column and the relative molecular wight was detected by SDS-PAGE as 3.0× 10^4 and 2.5× 10^4 for the two polysaccharide-protein complexes, It suggested that IC50 was 804.51 ug/mL by MTT. Glycoproteins A andd B were gained by Sephadex G-200 gel column from polysaccharide-protein complex. The IC50 of glycoproteins A and B were 532.96 and 613.91 ug/mL, respectively. Conclusion Material basis of cytotoxicity in polysaccharide from S. nigrum is two kinds of glycoproteins whose relative molecular weight are 3.0× 10^4 and 2.5× 10^4.