鼓槌石斛肠粘膜免疫活性多糖的筛选及其理化性质和结构组成的研究

发布时间：2018-07-15 18:42

【摘要】：本文以鼓槌石斛为材料,从肠道粘膜免疫活性出发研究鼓槌石斛的各个组分,通过将鼓槌石斛多糖的提取、分离、纯化和免疫活性筛选相结合,获得了均一的多糖组分(DCP-F4),优化了鼓槌石斛多糖的提取分离方法,对鼓槌石斛各组分进行了理化性质的测定,并对均一的高活性多糖的初级结构的进行分析,为鼓槌石斛多糖的构效关系的研究奠定了基础。实验结果如下:(1)均一活性多糖的获得:将鼓槌石斛干粉按照酒精醇提、热水浸提、醇沉、脱蛋白、透析的步骤依次提取、分离,对得到的提取物DCE-A(7.48%),DCE-B(7.93%),DCE-C(7.14%),DCE-D(4.67%),DCP(3.72%)进行肠免疫活性初步筛选。接着对DCP进行酒精分级醇沉、阴离子交换柱分离、葡聚糖凝胶柱分离结合活性筛选得到高活性组分DCP-F4。(2)鼓槌石斛多糖的肠道免疫活性:鼓槌石斛多糖的各提取物都具有很好体外肠道免疫活性,粗提取物作用组的骨髓细胞增殖率提高了3.3%-89.0%,通过逐级的分离,鼓槌石斛多糖含量越来越高,且肠道免疫活性也越来越高。粗多糖用60%的乙醇沉淀下来的多糖组分(DCP-2)的肠免疫活性较其他两个组分更好,在实验浓度范围内,骨髓细胞的增殖率在44.8%-91.4%。DCP-2进一步分离得到的各组分中以水洗糖的促进作用最强,骨髓细胞的增值率为56.1%-95.7%。水洗糖通过分离得到的组分中DCP-F4的活性最佳,对骨髓细胞的促进作用为对照组的2.136倍。(3)鼓槌石斛均一多糖理化性质测定:DCP-F4的碳水化合物含量为94.32%,糖醛酸含量为1.85%,重均分子量为1.37×104Da,多分散系数为1.1497。DCP-F4的傅里叶红外分析具有多糖的标志吸收峰。样品的DSC分析表明多糖仅有一个122℃吸收热和316℃放热峰。多糖的单糖组成是由Xyl、Man、Glc构成的,比例为依次为1:7:16;单糖组成分析结合核磁分析,它们的一级结构大致是以1,4-Linked Manp和1,4-Linked Glcp为主链,支链由1,4,6-Linked Manp分支形成,端基主要有T-Linked Xyl、T-Linked Glcp形成的糖链单元。
[Abstract]:In this paper, Dendrobium Dendrobii was used as the material to study the components of Dendrobium Dendrobium from the intestinal mucosal immune activity. By combining the extraction, separation, purification and immune activity screening of Dendrobium Dendrobium, the homogeneous polysaccharide component (DCP-F4) was obtained. The extraction and separation method of Dendrobium polysaccharide was optimized, and the components of Dendrobium Dendrobium were carried out. The determination of physical and chemical properties and the analysis of the primary structure of the homogeneous high active polysaccharide have laid a foundation for the study of the structure-activity relationship of Dendrobium polysaccharides. The results are as follows: (1) obtaining the homogeneous active polysaccharide: the steps of the dry powder of Dendrobium Dendrobium were extracted according to alcohol alcohol, hot water extraction, alcohol precipitation, deproteinization and dialysis. In addition, the intestinal immune activity of extracts DCE-A (7.48%), DCE-B (7.93%), DCE-C (7.14%), DCE-D (4.67%), DCP (3.72%) was preliminarily screened. Then alcohol fractionation and alcohol precipitation, anion exchange column separation, and dextran gel column separation and binding activity were screened to obtain the intestinal immune activity of Dendrobium Dendrobium polysaccharides with high active components DCP-F4. (2) (2): drums. The extracts from Dendrobium candidum polysaccharide have good in vitro intestinal immune activity. The proliferation rate of bone marrow cells in the crude extract group increased by 3.3%-89.0%. The polysaccharide content of Dendrobium Dendrobium was higher and higher, and the intestinal immune activity was higher and higher. The polysaccharide component (DCP-2) precipitated with 60% ethanol from the crude polysaccharide was immune to the intestinal immunity. The pestilence activity is better than that of the other two components. In the range of experimental concentration, the proliferation rate of bone marrow cells is the strongest in the components of 44.8%-91.4%.DCP-2 further separated by water washing sugar. The value added rate of bone marrow cells is the best activity of DCP-F4 in the components of 56.1%-95.7%. washed sugar through separation, and the promotion of bone marrow cells to the bone marrow cells. The effect was 2.136 times as the control group. (3) determination of physical and chemical properties of polysaccharide of Dendrobium Dendrobii: DCP-F4 carbohydrate content was 94.32%, glucuronic acid content was 1.85%, weight average molecular weight was 1.37 x 104Da, polydispersity coefficient was 1.1497.DCP-F4, Fourier infrared analysis had the mark absorption peak of polysaccharide. The DSC analysis of sample showed that the polysaccharide was only one The monosaccharide composition of polysaccharide is composed of Xyl, Man, Glc, and the proportion is 1:7:16, and the monosaccharide composition analysis combined with nuclear magnetic analysis. The first order structure of the monosaccharide is 1,4-Linked Manp and 1,4-Linked Glcp as the main chain, the branch chain is formed by 1,4,6-Linked Manp branch, and the end group mainly contains T-Linked Xyl. A sugar chain unit formed by Glcp.
【学位授予单位】：合肥工业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TQ281

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