多糖论文：迭鞘石斛所含多糖对小鼠免疫细胞影响的研究.doc

多糖论文：迭鞘石斛所含多糖对小鼠免疫细胞影响的研究

【中文摘要】石斛是我国常用的贵重药材,它具有益胃生津、滋阴清热、润肺止咳等功效。现代药理学研究发现,石斛不但具有治疗白内障、降血糖、增强人体免疫力的功能,还具有抗肿瘤、抗氧化、抗衰老和抗血栓等作用。石斛多糖作为石斛属植物重要的药用成分之一,已被证实具有增强免疫力的作用。为进一步探讨和研究石斛多糖对免疫力提高的作用机制,本研究采用正常小鼠免疫细胞体外培养的方法,通过添加提取自迭鞘石斛(Dendrobinm chryseum Rolfe.)的多糖成分,并给予LPS和ConA刺激,观察这类多糖对小鼠脾细胞和小鼠腹腔巨噬细胞增殖作用的影响以及对细胞因子的影响。(1)研究迭鞘石斛所含多糖对小鼠脾淋巴细胞以及腹腔巨噬细胞增殖的影响；(2)研究迭鞘石斛所含多糖对小鼠细胞因子的影响。方法：(1)分离提取小鼠脾淋巴细胞和腹腔巨噬细胞,通过添加和不添加迭鞘石斛所含多糖进行体外培养,以MTT法检测这些免疫细胞增殖的变化情况。(2)分离提取小鼠脾淋巴细胞和腹腔巨噬细胞,通过添加和不添加迭鞘石斛所含多糖进行体外培养,以ELISA法检测培养液中所含IL-2、IL-6的含量。结果：(1)迭鞘石斛所含多糖在体外对小鼠脾淋巴细胞增殖以及腹腔巨噬细胞的增殖有增强作用,说明对小鼠具有免疫增强作用。(2)通过迭鞘石斛多糖对小鼠细胞因子影响的实验发现,迭鞘石斛多糖对小鼠脾淋巴细胞在ConA刺激下分泌IL-2具有明显促进作用(p<0.01),并且具有量效关系；对小鼠腹腔巨噬细胞在LPS刺激下分泌IL-6也具有明显的促进作用(p<0.01),并且具有量效关系。结论：(1)通过迭鞘石斛多糖对小鼠免疫细胞增殖的实验发现,迭鞘石斛多糖对于小鼠脾淋巴细胞以及腹腔巨噬细胞都有一定的促进作用,低浓度时,作用不显著(2)通过迭鞘石斛多糖对小鼠细胞因子影响的实验发现,迭鞘石斛多糖对小鼠脾淋巴细胞在ConA刺激下分泌IL-2具有促进作用,并且具有量效关系；对小鼠腹腔巨噬细胞在LPS刺激下分泌IL-6也具有促进作用,并且具有量效关系。
【英文摘要】Dendrobium as an expensive medicinal herb, which has many important effects, has been widely used in China. Modern pharmacology study showed that Dendrobium has many therapeutic effects, such as improving cataract, antitumor, enhancing immunity, reducing blood sugar, antioxidation, antiaging and antithrombotic. Polysaccharides from Dendrobinm as one kind of important effective components in Dendrobinm has been proven to be able to enhance immunity. In order to further study the action mechanism of Dendrobinm polysaccharides on immunity, this study investigated the effects of polysaccharides from Dendrobinm chryseum Rolfe on growthing of mice spleen cells and peritoneal macrophages stimulated by LPS and ConA with MTT, and on expression of some cytokines of the cells with ELISA.:(1) To study the effects of the polysaccharides extracted from Dendrobinm chryseum Rolfe on proliferation of mice spleen lymphocytes and peritoneal macrophages. (2) To research the influences of the polysaccharides extracted from Dendrobinm chryseum Rolfe on IL-2, IL-6 of mice spleen lymphocyte and peritoneal macrophages.Method:(1) The mice spleen cells and peritoneal macrophages were cultured in vitro. They were stimulated by LPS and ConA. They were treated with or without the polysaccharide. Their proliferations were detected with MTT. (2) The mice spleen cells and peritoneal macrophages were cultured in vitro. They were treated with or without the polysaccharide. Their IL-2, IL-6 of the culture medium were detected with ELISA.Conclusion:(1) The polysaccharide from Dendrobinm chryseum Rolfe can promote the proliferation of mice spleen lymphocytes and peritoneal macrophages in vitro (p<0.01). It is able to enhance the immunity of mouse. (2) The polysaccharide from Dendrobinm chryseum Rolfe can promote the expression of IL-2, IL-6 of spleen lymphocytes and peritoneal macrophages stimulated by LPS or ConA (p<0.01). These effects had the concentration-response relations.
【关键词】多糖 脾淋巴细胞 腹腔巨噬细胞 迭鞘石解 细胞因子
【英文关键词】Polysaccharide Spleen lymphocyte Peritoneal macrophage Dendrobinm chryseum Rolfe Cytokine
【目录】迭鞘石斛所含多糖对小鼠免疫细胞影响的研究

摘要

6-7

Abstract

7

第1章 绪论

10-29

1.1 石斛

10-15

1.1.1 石斛简介

10

1.1.2 常见的石斛种类

10-13

1.1.3 石斛的主要药用成分

13-14

1.1.4 石斛的药用价值

14-15

1.2 石斛多糖

15-17

1.2.1 石斛多糖的提取工艺

15-16

1.2.2 多糖的结构

16-17

1.3 石斛多糖的药理学研究

17-19

1.3.1 抗肿瘤作用

17

1.3.2 增强免疫系统作用

17-19

1.3.3 降血糖作用

19

1.4 迭鞘石斛

19-21

1.4.1 迭鞘石斛的分布

19

1.4.2 迭鞘石斛的鉴定

19-20

1.4.3 迭鞘石斛的化学成分

20-21

1.5 迭鞘石斛药理作用

21-22

1.6 免疫细胞

22-24

1.6.1 巨噬细胞

22

1.6.2 淋巴细胞

22-24

1.7 细胞因子

24-29

1.7.1 白细胞介素

25-26

1.7.2 集落刺激因子

26-27

1.7.3 干扰素

27

1.7.4 肿瘤坏死因子

27-28

1.7.5 淋巴因子

28

1.7.6 单核因子

28-29

第2章 迭鞘石斛多糖对小鼠免疫细胞增殖的影响

29-44

2.1 迭鞘石斛多糖对小鼠脾淋巴细胞的影响

29-37

2.1.1 实验材料

29-30

2.1.2 实验步骤

30-36

2.1.3 实验结果

36-37

2.2 石斛多糖对小鼠腹腔巨噬细胞的影响

37-42

2.2.1 实验材料

37-39

2.2.2 实验步骤

39-40

2.2.3 实验结果

40-42

2.3 讨论

42-44

第3章 迭鞘石斛多糖对小鼠细胞因子的影响

44-54

3.1 实验材料

44-45

3.1.1 实验动物

44

3.1.2 实验试剂

44-45

3.1.3 实验仪器

45

3.2 实验方法

45-52

3.2.1 ELISA法测定小鼠脾淋巴细胞IL-2含量水平

45-48

3.2.2 ELISA法测定小鼠腹腔巨噬细胞IL-6含量水平

48-52

3.3 讨论

52-54

结论

54-55

致谢

55-56

参考文献

56-60

攻读硕士学位期间发表的论文

60