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为了研究提取溶剂酸碱性对柳蘑多糖的制备及抑制肿瘤细胞增殖的影响,本文利用稀酸、蒸馏水和稀碱溶剂分别制备三种柳蘑多糖(SPAP1、WPAP1和JPAP1),经过脱蛋白对应得到三种脱蛋白的柳蘑多糖(SPAP2、WPAP2和JPAP2),分别考察柳蘑多糖提取率和纯度的变化以及对肿瘤细胞增殖抑制作用的影响,并采用红外光谱仪初步分析柳蘑多糖的结构差异。研究表明,提取溶剂酸碱性对柳蘑多糖的提取率及纯度均有显著影响,其中JPAP1的提取率最高,为11.02%,SPAP2的纯度最高,为73.91%。SPAP2对肿瘤细胞增殖抑制效果显著高于其它柳蘑多糖(p<0.05),并与多糖浓度和给药时间成量效关系。其中SPAP2对Hela细胞的增值抑制效果最好,抑制率为90.88%;红外光谱分析SPAP2在1078±4 cm-1和759.83±5 cm-1处有较强的吸收峰,表明SPAP2中C-O-H或C-O-C和环氧基团含量高于其它柳蘑多糖。本论文结果为深入开发柳蘑多糖产品及进一步研究柳蘑多糖诱导肿瘤细胞凋亡机制提供理论依据。
In order to study the effects of extraction solvent pH and alkalinity on the preparation and the inhibition of the proliferation of the tumor cells, three kinds of polysaccharides (SPAP1, WPAP1 and JPAP1) were prepared by dilute acid, distilled water and dilute alkaline solvent respectively. Three kinds of deproteinized willow polysaccharides (SPAP2, WPAP2 and JPAP2) were obtained. The changes of the extraction rate and purity of the polysaccharides from Willow mushroom and the effects on the proliferation of the tumor cells were studied. The structural differences of the polysaccharides from the willow bromeis were analyzed by infrared spectroscopy . The results showed that the extraction solvent acidity and alkalinity had significant effects on the extraction rate and purity of the polysaccharide from B. juncea. The highest extraction rate of JPAP1 was 11.02% and the highest purity of SPAP2 was 73.91%. The inhibitory effect of SPAP2 on tumor cell proliferation was significantly higher than that of other B. juncea polysaccharides (p <0.05), and it was dose-effect relationship with polysaccharide concentration and administration time. Among them, SPAP2 had the best inhibitory effect on the proliferation of Hela cells with the inhibition rate of 90.88%. The SPAP2 had strong absorption peaks at 1078 ± 4 cm-1 and 759.83 ± 5 cm-1 by FTIR, which indicated that COX or COC And epoxy content higher than other willow mushroom polysaccharides. The results of this dissertation provide a theoretical basis for the further development of Salicaceae polysaccharide products and further research on the mechanism of Salvia miltiorrhiza polysaccharides inducing tumor cell apoptosis.