党参多糖单糖组成与其对HepG2细胞毒活性的相关分析

Monosaccharide compositions of Codonopsis pilosula polysaccharides and their correlation analysis on cytotoxic activities against HepG2 cells

目的探究党参多糖的单糖组成与其对肝癌HepG2细胞毒活性之间的相关性。方法 26批不同产地的党参样品,采用水提醇沉法提取多糖,苯酚硫酸法测多糖的量、气相色谱法和间羟基联苯法同时测定半乳糖醛酸的量、糖腈乙酸酯衍生化法分析单糖种类及量、三甲基硅醚衍生化法测果糖的量,MTT法研究党参多糖对HepG2细胞的细胞毒活性,采用聚类分析法对26批党参样品进行聚类分析,以偏最小二乘法(PLS)探究样品中各单糖种类和量与其对HepG2细胞毒活性的相关性。结果 26批党参多糖样品均显示一定的HepG2细胞毒活性,且以甘肃文县的8号党参多糖的细胞毒活性最强(其抑制率为36.36%)。26批党参多糖中各类单糖的量存在差异。聚类分析结果表明,党参多糖的单糖种类及量不能作为党参分类的指标。党参多糖的单糖组成与HepG2细胞毒活性的相关性研究表明,半乳糖醛酸、鼠李糖、阿拉伯糖、半乳糖、果糖与HepG2细胞毒活性呈正相关,而甘露糖、木糖和葡萄糖与HepG2细胞毒活性呈负相关。结论党参多糖对HepG2的细胞毒活性与其单糖的种类存在相关性,含有半乳糖醛酸相对较高的党参多糖的细胞毒活性较强。

Objective To study the correlation between monosaccharide compositions and their in vitro cytotoxic activities. Methods Twenty-six Codonopsis pilosula polysaccharides （CPPs） were extracted by water extraction and alcohol precipitation method from C. pilosula collected from 26 different habitats. Total carbohydrate contents of the polysaccharides were determined by the phenol-sulfuric acid colorimetric method. The methods of m-hydroxydiphenyl and GC were all applied to the determination of the galacturonic acid contents in polysaccharides, and the compositional analysis was performed using the aldononitrile acetate method while the fructose contents was analyzed by the trimethylsilyl ether method. In vitro cytotoxic activities against human hepatocellular carcinoma HepG2 cells of the 26 batches of CPPs were evaluated by MTT. Then, hierarchical clustering analysis （HCA） was used to study the classification of 26 batches of C.pilosula and partial least squares （PLS） was used to investigate the correlation between the monosaccharide compositions and in vitro cytotoxic activities of the polysaccharides. Results The results showed that all the 26 batches of CPPs possessed the cytotoxic activities against HepG2 cells. The 8th CPPs sample, which was extracted from the C. pilosula collected in Wen county （Gansu province, China）, had the highest growth inhibitory rate on HepG2 cells and the inhibitory rate was 36.36%. In addition, the monosaccharide contents of the 26 batches of CPPs were different from each other. The results of HCA showed that the C. pilosula could not be classified according to the monosaccharide compositions. The PLS results indicated that galacturonic acid, arabinose, rharnnose, galactose, and fructose were correlated positively with the cytotoxic activities, while rnannose, xylose, and glucose were correlated negatively with the cytotoxic activities. Conclusion The in vitro cytotoxic activities of CPPs are highly correlative to their monosaccharide compositions, and the CPPs containing much galacturonic acid have the obviously cytotoxic activities.