柴达木唐古特白刺(Nitraria tangutorum Bobr.)活性多糖的研究
Alternative TitleStudy on Activite Polysaccharide of Nitraria tangutorum Bobr. in Chaidam
Thesis Advisor索有瑞
Degree Grantor中国科学院西北高原生物研究所
Place of Conferral西北高原生物研究所
Keyword唐古特白刺 多糖 提取工艺 单糖组成 分子量 矿质元素 调节免疫 清除自由基
Abstract白刺为蒺藜科(Zygophyllceae)白刺属(Nitraria)植物的总称,是一种落叶灌木。在柴达木盆地有三个种的分布,其中唐古特白刺(Nitraria tangutorum Bobr.)为优势种。本论文选择唐古特白刺的果实作为实验材料,对果实中的多糖成分进行了较为全面、系统的研究,主要内容包括:优化了白刺多糖的提取工艺;通过DEAE-52和Sephadex G-100柱层析分离得到7种分子量均一的多糖组分,并进行了初步的结构鉴定;对白刺多糖的免疫力调节作用、体外抗氧化活性进行了初步探索。具体实验结果如下: 1、通过正交实验探讨了白刺多糖的最佳提取工艺。在多糖提取前要进行脱脂,除单糖等预处理工作。实验结果表明:白刺多糖的最佳提取工艺是料液比为1:5,94℃回流提取3次,每次2h。在此条件下得到的白刺多糖含量为34.56mg/g,提取得率为48.2%。 2、采用Sevage法脱蛋白,X-5大孔吸附树脂除色素对白刺粗多糖进行初步纯化得到精制粗多糖,并利用DEAE-52、Sephadex G-100柱层析进一步分离多糖,从中得到7种分子量均一的多糖组分,分别命名为NIPⅠ、NIPⅡ、NIPⅢ、NIPⅣ、NIP Ⅴ、NIP Ⅵ、NIP Ⅶ。 3、对白刺多糖进行了初步的结构鉴定,包括:通过高效毛细管区带电泳(HPCE)进行单糖组成分析;高效凝胶渗透色谱法(HPGPC)测分子量;红外光谱测定。对四种不同纯度级别的白刺多糖的理化性质检测表明,经过多次分离纯化步骤得到的分子量均一的纯品多糖是不含蛋白质、游离单糖和酚类的非淀粉类多糖。 4、测定了唐古特白刺粗多糖中17种微量元素(Cu、Zn、Fe、Mn、Co、Ni、Se、Cr、Mg、Ca、K、Na、P、As、Hg、Pb、Cd)。结果表明:唐古特白刺粗多糖中含有的必需常量元素K、Ca、Na含量较高;人体必需微量元素Fe、Zn、Mn的含量均较高,尤其以Fe的含量为高;非必需微量元素(有害元素)As、Hg、Pb、Cd的含量低于1mg/kg。 5、以环磷酰胺致动物免疫功能低下,建立免疫实验模型,研究白刺果实粗多糖的免疫调节作用。实验结果表明,白刺粗多糖0.70g/kg灌胃剂量能显著增强模型小鼠单核巨噬细胞吞噬功能;0.35g/kg和0.70g/kg灌胃剂量能显著提高模型小鼠迟发型超敏反应;0.35g/kg、0.70g/kg和1.4g/kg灌胃剂量能显著提高模型小鼠血清溶血素生成能力。 6、采用超氧阴离子自由基体系(O2-•)、二苯代苦味酰基自由基体系(DPPH•)和羟自由基体系(OH•),对白刺多糖的体外抗氧化活性进行研究,实验结果显示,白刺多糖对这三种自由基均有不同程度的清除作用。将白刺粗多糖、白刺精制多糖同L-抗坏血酸进行比较,结果显示:对超氧阴离子自由基的清除能力为L-抗坏血酸>白刺精制多糖>白刺粗多糖;对DPPH自由基的清除能力为白刺粗多糖>L-抗坏血酸>白刺精制多糖;对羟自由基的清除能力为L-抗坏血酸>白刺精制多糖>白刺粗多糖。 本论文通过对白刺果实活性多糖的系统研究,为开发白刺多糖的相关产品提供理论基础,为白刺果实的综合开发利用提供科学依据。
Other AbstractNitraria is a kind of shrub which belongs to the family of Zygophyllceae. There are 3 species in Chaidam Basin. Nitraria tangutorum Bobr. distribute in large quantities there as the dominant species. In this research, the polysaccharide in the fruit of Nitraria tangutorum Bobr. has been studied systematically, which including the extraction techniques of polysaccharides, the purification techniques of seven relatively homogeneous polysaccharides through DEAE-52 and Sephadex G-100 column chromatography, the structure identification, immune and anti-oxidative activities of polysaccharides. Some results and conclusions were obtained as follows. 1. The optimum extracting conditions of Nitraria polysaccharide (NIP) were investigated by orthogonal experiment. Fat and monosaccharide were removed before the extraction of polysaccharide. The results showed under the conditions of 94 ℃, 1:5(W/V), extracting 3 times and 2 h for each time, the content of polysaccharide was 34.56 mg/g and the ratio was 48.2%. 2. After removing protein by the method of sevage and decolorization with X-5 macroporous resin, seven polysaccharide components NIPⅠ, NIPⅡ, NIPⅢ, NIPⅣ, NIPⅤ, NIPⅥ, NIPⅦ have been separated through DEAE-52 and Sephadex G-100 column chromatography from the crude Nitraria polysaccharide. The peak shape showed that they were relatively homogeneous polysaccharides. 3. The structures of seven relatively homogeneous polysaccharides were studied. The monosaccharide compositions and the molecular weight have been determined by capillary zone electrophoresis and high performance gel permeation chromatographic method, respectively. Their IR spectra were also studied. Physicochemical characteristics analysis indicated that the relatively homogeneous polysaccharides did not contain protein, free monosaccharide, polyphenols and starch. 4. The contents of 17 mineral elements (Cu、Zn、Fe、Mn、Co、Ni、Se、Cr、Mg、Ca、K、Na、P、As、Hg、Pb、Cd) in crude polysaccharides of Nitraria tangutorun Bobr. were analyzed. The results show that the contents of K, Ca, Na were very abundant. The contents of essential trace elements Fe, Zn, Mn were also abundant, in which Fe was especially high. And the contents of hazard elements As、Hg、Pb、Cd were low, which were all less than 1mg/kg. 5. In this research, the immunomodulatory effect of NIP was studied on cyclophosphamide (CPA) model mice. The experimental results indicated that NIP ig 0.70 g/kg remarkably strengthened phagocytosing function of the monocyte- macrophage system in mice treated by CPA; NIP ig 0.35 g/kg or 0.70 g/kg remarkably enhanced delayed type hypersensitivity reaction of the experimental mice; NIP ig 0.35 g/kg or 0.70 g/kg or 1.4 g/kg significantly improved the production of serum hemolysin in model mice. 6. The free radical scavenging activities of NIP in vitro were studied using superoxide anion (O2-•), radical and hydroxyl radical (OH•). The results indicated that NIP had different scavenging effects on these three free radicals. The scavenging capacities of crude polysaccharide, refined polysaccharide and Vitamin C for O2-•、DPPH• and OH• were compared. The orders of scavenging capacity of them were as follow: Vitamin C > refined polysaccharide> crude polysaccharide for O2-•; crude polysaccharide>Vitamin C >refined polysaccharide for DPPH•; Vitamin C > refined polysaccharide> crude polysaccharide for OH•. The systematical research of NIP can provide the academic foundation for developing products about NIP and supply the scientific basis for reasonable usage of the Nitraria resource.
Document Type学位论文
Recommended Citation
GB/T 7714
王凌云. 柴达木唐古特白刺(Nitraria tangutorum Bobr.)活性多糖的研究[D]. 西北高原生物研究所. 中国科学院西北高原生物研究所,2009.
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