板栗种仁多糖的提取纯化及体外抗肿瘤活性筛选

以燕山地区板栗为原料,利用加压溶剂萃取法提取板栗种仁粗多糖,以多糖得率为指标,通过单因素结合响应面法优化提取工艺条件。将最优提取条件下得到的粗多糖经除蛋白、脱色素、透析后得到纯化多糖,继而借助制备型高效体积排阻色谱得到板栗多糖的高分子量组分(YCP-H),分析了其形貌特点、结构特征和单糖组成,并针对8种人体肿瘤细胞模型进行了体外抗肿瘤活性筛选。结果表明,高压溶剂萃取法提取板栗种仁粗多糖的最佳提取条件为:当投料量为10 g,静态提取次数为3次时,设定温度60 ℃、时间9 min、压力6 MPa,多糖得率可达19.78%±0.27%。组分YCP-H是具有β-吡喃构型的酸性多糖,重均分子量范围为217~5900 kDa,在扫描电镜下呈现纤维状的微观形貌,单糖组成包含阿拉伯糖、半乳糖、葡萄糖、木糖、甘露糖和岩藻糖。YCP-H对人肝癌细胞HepG2的增殖具有显著(P<0.05)的抑制作用,其半数抑制浓度可达0.08 μg/mL。此外,YCP-H对人结肠癌细胞HCT-116和肺癌细胞A-549的增殖也有较强的抑制作用。

Extraction,Purification and in Vitro Antitumor Screening of Polysaccharides from Chestnut Kernel

Crude polysaccharides in chestnut kernel from Yanshan area were extracted by high-pressure solvent extraction. In terms of the extraction yield of polysaccharide, single factor tests and Box-Behnken design-response surface method were applied to optimize the extraction conditions. The crude polysaccharides extracted under the optimum conditions was purified through deproteinization, depigmentation and dialysis to obtain purified polysaccharides, and then the high molecular weight component (YCP-H) was separated by the preparative high-performance size-exclusion chromatography. The morphological characteristics, structural features and monosaccharide composition of YCP-H were analyzed, and a preliminary assay on in vitro antitumor activity against eight selected cancer cell lines was also carried out. The results showed that the optimum extraction conditions for crude polysaccharides were extraction temperature of 60℃, static extraction time of 9 min and extraction pressure of 6 MPa when the amount of raw materials was 10 g and cycle times was 3, with a corresponding maximum extraction yield of 19.78%±0.27%. The component YCP-H was an acidic polysaccharide which had β-pyranose configuration. YCP-H had weight-average molecular weight covering the range of 217~5900 kDa, and exhibited fiber-like morphology under scanning electron microscope. The monosaccharide composition analysis showed that it was composed of arabinose, galactose, glucose, xylose, mannose and fucose. YCP-H had significant anti-proliferative effects on human hepatoma HepG2 cells, with a half-maximal inhibitory concentration of 0.08 μg/mL. In addition, YCP-H could also effectively inhibit the proliferation of HCT-116 human colon cancer cell line and A-549 adenocarcinoma epithelial cell line.