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霍山石斛是名贵中药材,其多糖的抗肿瘤活性是中药资源领域的研究热点,该研究旨在探究霍山石斛多糖抗肿瘤活性的物质基础。实验以霍山石斛为原料,通过低盐溶液浸提和硫酸铵分级沉淀得到不同的霍山石斛蛋白组分,经SDS-PAGE电泳染色确定糖蛋白组分,再经DEAE离子柱和Sephadex凝胶柱进一步分离纯化糖蛋白组分,同时采用MTT比色法检测不同产物对人肝癌细胞Hep G2的毒活性;结果获得3种相对分子质量为22.5,19.8,15.6 k Da的糖蛋白组分RG1,RG2,RG3,测得三者复合物对Hep G2的IC50为534.23 mg·L-1,而组分RG1,RG2的IC50分别为432.96,413.91 mg·L-1,组分RG3对Hep G2无毒活性。总之,实验结果提示霍山石斛多糖抗肿瘤活性的物质基础之一是相对分子质量为22.5,19.8 k Da的2种糖蛋白组分,且具有协同效用。
Dendrobium huoshanense is a valuable Chinese herbal medicine, and its antitumor activity of polysaccharides is a hot spot in the field of traditional Chinese medicine resources. The purpose of this study is to explore the material basis of antitumor activity of Dendrobium huoshanense. Dendrobium huoshanense was used as raw material to obtain different components of Dendrobium huoshanense by low salt solution extraction and ammonium sulfate fractionation. The glycoprotein components were determined by SDS-PAGE electrophoresis and further purified by DEAE and Sephadex columns The glycoprotein components were isolated and purified. At the same time, the toxicity of different products to Hep G2 cells was detected by MTT colorimetric assay. Three glycoproteins RG1, RG2, RG3. The IC50 values of the three compounds against Hep G2 were 534.23 mg · L-1, while the IC50 values of RG1 and RG2 were 432.96 and 413.91 mg · L-1, respectively. RG3 had no toxicity to Hep G2. In conclusion, one of the material foundations for the antitumor activity of polysaccharides from Dendrobium huoshanense was the two glycoproteins with relative molecular masses of 22.5 and 19.8 kDa and synergistic effects.