【摘要】：本文以鼓槌石斛為材料,從腸道粘膜免疫活性出發(fā)研究鼓槌石斛的各個組分,通過將鼓槌石斛多糖的提取、分離、純化和免疫活性篩選相結(jié)合,獲得了均一的多糖組分(DCP-F4),優(yōu)化了鼓槌石斛多糖的提取分離方法,對鼓槌石斛各組分進行了理化性質(zhì)的測定,并對均一的高活性多糖的初級結(jié)構(gòu)的進行分析,為鼓槌石斛多糖的構(gòu)效關(guān)系的研究奠定了基礎(chǔ)。實驗結(jié)果如下:(1)均一活性多糖的獲得:將鼓槌石斛干粉按照酒精醇提、熱水浸提、醇沉、脫蛋白、透析的步驟依次提取、分離,對得到的提取物DCE-A(7.48%),DCE-B(7.93%),DCE-C(7.14%),DCE-D(4.67%),DCP(3.72%)進行腸免疫活性初步篩選。接著對DCP進行酒精分級醇沉、陰離子交換柱分離、葡聚糖凝膠柱分離結(jié)合活性篩選得到高活性組分DCP-F4。(2)鼓槌石斛多糖的腸道免疫活性:鼓槌石斛多糖的各提取物都具有很好體外腸道免疫活性,粗提取物作用組的骨髓細胞增殖率提高了3.3%-89.0%,通過逐級的分離,鼓槌石斛多糖含量越來越高,且腸道免疫活性也越來越高。粗多糖用60%的乙醇沉淀下來的多糖組分(DCP-2)的腸免疫活性較其他兩個組分更好,在實驗濃度范圍內(nèi),骨髓細胞的增殖率在44.8%-91.4%。DCP-2進一步分離得到的各組分中以水洗糖的促進作用最強,骨髓細胞的增值率為56.1%-95.7%。水洗糖通過分離得到的組分中DCP-F4的活性最佳,對骨髓細胞的促進作用為對照組的2.136倍。(3)鼓槌石斛均一多糖理化性質(zhì)測定:DCP-F4的碳水化合物含量為94.32%,糖醛酸含量為1.85%,重均分子量為1.37×104Da,多分散系數(shù)為1.1497。DCP-F4的傅里葉紅外分析具有多糖的標(biāo)志吸收峰。樣品的DSC分析表明多糖僅有一個122℃吸收熱和316℃放熱峰。多糖的單糖組成是由Xyl、Man、Glc構(gòu)成的,比例為依次為1:7:16;單糖組成分析結(jié)合核磁分析,它們的一級結(jié)構(gòu)大致是以1,4-Linked Manp和1,4-Linked Glcp為主鏈,支鏈由1,4,6-Linked Manp分支形成,端基主要有T-Linked Xyl、T-Linked Glcp形成的糖鏈單元。
[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.
【學(xué)位授予單位】：合肥工業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：TQ281

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