摘 要：目的：采用麸皮多糖（WBR）刺激巨噬细胞RAW264.7，建立体外的炎症模型，研究WBR对巨噬细胞免疫因子释放的影响。方法:红外光谱法测定WBR的结构；运用气相色谱法测定WBR的单糖的种类；不同浓度麸皮多糖刺激RAW264.7细胞,采用MTT法检测不同浓度的样品对RAW264.7细胞毒性作用；采用Griess法检测细胞液中NO的含量；采用ELISA法检测细胞液中PGE2、TNF-含量。结果：随着WBR浓度的增加细胞存活率无明显的差异；并且随着WBR的浓度的增加细胞液中NO的含量和PGE2、TNF-含量也随之增加。结论：WBR具有促进炎症相关因子表达量，从而增加促炎症反应。71516

毕业论文关键词:麸皮多糖，提取，分离纯化，巨噬细胞，细胞免疫因子

Abstract: Purpose: WBR was used to stimulate macrophage RAW264. 7, also inflammation model in vitro was built in order to study the effects of WBR to the release of macrophage immune factor. Methods: The structure of WBR was measured by Infrared spectroscopy; The species of WBR’s monosaccharide was identified by gas chromatographic method; RAW264.7 cells were stimulated with different concentrations of bran polysaccharide, then the toxic effects on different concentrations of samples to RAW264.7 cell were determined by MTT; The method of Griess was used to detect the content of NO in the cell SAP; Using the ELISA method to detect the contents of PGE2 and TNF- in the cell SAP. Results: There was no difference in the rate of cell survival, along with the increase of concentrations of WBR ;Also, the content of NO、 PGE2 and TNF -in cell SAP increased with the increase of the concentration of the WBR. Conclusion: the WBR contributed to promote the expression index of the factors which were related to inflammation, thus increasing proinflammatory response.

Key words: bran polysaccharide, Extraction, Separation and purification, Macrophages, Cellular immune factor

目  录

1 引言 6

1.1 炎症的研究进展 6

1.1.1 炎症 6

1.1.2 参与炎症反应细胞 7

1.1.3 诱导炎症的细胞因子 7

1.2 多糖的概述 8

1.2.1 多糖 8

1.2.2 麸皮多糖 8

1.3 麸皮多糖的提取分离与研究方法 9

1.3.1 麸皮多糖的提取分离 9

1.3.2 麸皮多糖的纯化 9

1.3.3 红外光谱分析 10

1.3.4 气相色谱分析 10

2 材料与方法 10

2.1 实验材料与仪器 10

2.1.1 实验材料 10

2.1.2 实验仪器 11

2.2 方法 11

2.2.1 小麦麸皮多糖样品预处理 11

2.2.2 小麦麸皮粗多糖精制 11

2.2.3 多糖红外光谱分析 11

2.2.4 多糖单糖组分分析 12

2.2.5 RAW264.7细胞培养 12

2.2.6 RAW264.7的复苏 12

2.2.7 RAW264.7的传代 12

2.2.8 不同浓度样品对RAW264.7细胞毒性的检测