新疆党参多糖的抗肿瘤作用研究
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摘要
目的:新疆党参(Codonopsis Clematidea Clarke)为桔梗科党参属多年生草本植物,在全疆均有分布,且储量丰富,但其利用却很少。本课题期望通过对新疆党参多糖(Codonopsis Clematidea Clarke polysaccharide ,CCP)理化性质、抗肿瘤作用等的研究,探讨CCP作为一种抗癌新药的临床应用前景,为开发新的抗癌药物提供实验依据。
方法:(1)用水提醇沉法从生药新疆党参中得到粗多糖,Sevag法反复萃取5次去除多糖中混合的蛋白质,用丙酮、乙醚脱水洗涤4次,冷冻干燥后透析袋透析得多糖。多糖经过Sephadex G-75凝胶柱纯化,得到较纯的的多糖样品。经过溶解性、Molish试验、Fehling试验等实验的鉴定,确定所得到成分的为多糖;采用苯酚-硫酸比色法检测多糖的含量。(2)采用小鼠肉瘤S180移植模型,对CCP的体内抑瘤率、荷瘤小鼠的生命延长率进行测定。通过脾脏指数、以及ELISA法检测血清TNF-α水平,观察CCP对免疫系统的影响。(3)从昆明小鼠腹腔分离巨噬细胞,进行原代培养,检测CCP对巨噬细胞吞噬活性的影响。
结果:(1) CCP样品中多糖含量为36.01 %。(2)新疆党参多糖对荷瘤小鼠的抑瘤率为54%,生命延长率为17.7%,并且明显增加荷瘤小鼠的脾脏指数,升高血清TNF-α水平。(3) CCP在浓度30 mg/mL、80 mg/ mL、128 mg/mL均可以明显增强巨噬细胞的吞噬活性。
结论:(1)CCP样品中多糖含量为36.01 %,新疆党参中多糖含量很高,具有很好的开发前景。(2)CCP具有一定的抗肿瘤作用,且与化疗药物合用具有增效作用。(3)CCP可以增加荷瘤小鼠的脾脏指数,升高荷瘤小鼠血清TNF-α水平,增强巨噬细胞的吞噬活性,说明调节机体的免疫功能可能是抗肿瘤作用的机理之一。(4)CCP在治疗癌性恶病质方面具有研究价值。
Object: Codonopsis Clematidea Clarke in Xinjiang is the platicodon grandiflorum branch codonopsis pilosula.. It is distributed in Xinjiang province, and the reserves are abundant, In order to offer the basis for developing and utilizing of Codonopsis Clematidea Clarke. We did research about the medicine’s physicochemical properties and anti-tumor activity .
Methods:(1)The polysaccharide was isolated by extraction with hot water and precipated with alcohol. In order to remove the mixed protein, the Sevag extract for 5 times. Polysaccharide had been washed 4 times with acetone and ether dehydration, after freeze-drying and dialysis. We got pure polysaccharide sample after Sephadex G-75 gelatin column purification. With absorbance experiment,Molish experiment and Fehling experiment , etc,. The sample we got whose ingredient is the polysacch- aride. We got the polysaccharide content through the phenol - sulfuric acid color method;(2) The anti-tumor effects of CCP were tested in mice with transplanted tumorsarcoma-180,the tumor inhibition rate, life span , speen index and level of serm TNF-αare determined.(3)peritoneal macrophage were isolated from KM mice, phagocytosis of macrophage was determined in the concentration of 30 mg/m L,80 mg/ m L,128 mg/m L.
Result:(1) the CCP content is 36.01 %., (2) the tumor inhibitory rate of CCP to S180 was 54%,life prolong e rate is 17.7%,The spleen index and level of serm TNF-αin the groups treated with CCP were higher than control group(3)CCP can enhance the phagocytosis activity of macrophage greatly in the concentration of 30 mg/mL、80 mg/ m L、128 mg/m L.
Conclusion: (1)the CCP content is high which will have very good prospect.(2)CCP has effective anti-tumor activity and can enhance the anti-tumor effect of CTX(3) The anti-tumor role of CCP can be induced by ehancing immune function.(4)CCP is worth to continue to be researched in cancer cachexia.
方法:(1)用水提醇沉法从生药新疆党参中得到粗多糖,Sevag法反复萃取5次去除多糖中混合的蛋白质,用丙酮、乙醚脱水洗涤4次,冷冻干燥后透析袋透析得多糖。多糖经过Sephadex G-75凝胶柱纯化,得到较纯的的多糖样品。经过溶解性、Molish试验、Fehling试验等实验的鉴定,确定所得到成分的为多糖;采用苯酚-硫酸比色法检测多糖的含量。(2)采用小鼠肉瘤S180移植模型,对CCP的体内抑瘤率、荷瘤小鼠的生命延长率进行测定。通过脾脏指数、以及ELISA法检测血清TNF-α水平,观察CCP对免疫系统的影响。(3)从昆明小鼠腹腔分离巨噬细胞,进行原代培养,检测CCP对巨噬细胞吞噬活性的影响。
结果:(1) CCP样品中多糖含量为36.01 %。(2)新疆党参多糖对荷瘤小鼠的抑瘤率为54%,生命延长率为17.7%,并且明显增加荷瘤小鼠的脾脏指数,升高血清TNF-α水平。(3) CCP在浓度30 mg/mL、80 mg/ mL、128 mg/mL均可以明显增强巨噬细胞的吞噬活性。
结论:(1)CCP样品中多糖含量为36.01 %,新疆党参中多糖含量很高,具有很好的开发前景。(2)CCP具有一定的抗肿瘤作用,且与化疗药物合用具有增效作用。(3)CCP可以增加荷瘤小鼠的脾脏指数,升高荷瘤小鼠血清TNF-α水平,增强巨噬细胞的吞噬活性,说明调节机体的免疫功能可能是抗肿瘤作用的机理之一。(4)CCP在治疗癌性恶病质方面具有研究价值。
Object: Codonopsis Clematidea Clarke in Xinjiang is the platicodon grandiflorum branch codonopsis pilosula.. It is distributed in Xinjiang province, and the reserves are abundant, In order to offer the basis for developing and utilizing of Codonopsis Clematidea Clarke. We did research about the medicine’s physicochemical properties and anti-tumor activity .
Methods:(1)The polysaccharide was isolated by extraction with hot water and precipated with alcohol. In order to remove the mixed protein, the Sevag extract for 5 times. Polysaccharide had been washed 4 times with acetone and ether dehydration, after freeze-drying and dialysis. We got pure polysaccharide sample after Sephadex G-75 gelatin column purification. With absorbance experiment,Molish experiment and Fehling experiment , etc,. The sample we got whose ingredient is the polysacch- aride. We got the polysaccharide content through the phenol - sulfuric acid color method;(2) The anti-tumor effects of CCP were tested in mice with transplanted tumorsarcoma-180,the tumor inhibition rate, life span , speen index and level of serm TNF-αare determined.(3)peritoneal macrophage were isolated from KM mice, phagocytosis of macrophage was determined in the concentration of 30 mg/m L,80 mg/ m L,128 mg/m L.
Result:(1) the CCP content is 36.01 %., (2) the tumor inhibitory rate of CCP to S180 was 54%,life prolong e rate is 17.7%,The spleen index and level of serm TNF-αin the groups treated with CCP were higher than control group(3)CCP can enhance the phagocytosis activity of macrophage greatly in the concentration of 30 mg/mL、80 mg/ m L、128 mg/m L.
Conclusion: (1)the CCP content is high which will have very good prospect.(2)CCP has effective anti-tumor activity and can enhance the anti-tumor effect of CTX(3) The anti-tumor role of CCP can be induced by ehancing immune function.(4)CCP is worth to continue to be researched in cancer cachexia.
引文
1. 向道斌,李晓玉.多糖的免疫调节作用[J].国外医学-合成药、生化药、制剂分册,1991,12(5):261-267.
2. 李春德译. 一种新的治疗药物的来源--糖类的研究[J]. 国外医学 药学分册,1994,21(6):353-356.
3. 李铁民,梁再赋. 甘草提取物及其衍生物的抗病毒研究现状[J].中草药,1994,25(12):655-658.
4. 张松,徐章.多糖类医药生物活性研究进展[J].中国生化药物志,1996,17(6):272-275.
5. 赵勇,张玲. 中药免疫调节研究进展[J]. 中草药,1994,25(11):603-606.
6. 丁媛媛. 多糖与抗病毒作用. 国外医学-微生物学分册,2001,24(1):18-20.
7. 李建恒,张杏红. 抗肿瘤中药多糖研究进展.中医药学报,1998,26(4):46-48.
8. Srubastava R,Kulshreshtha D.K. Biocide polysaccharides from plants. Phytochemistry, 1989,28(11):2877-2883.
9. 方积年.多糖研究的现状. 药学学报,1986,21(12):944-948.
10. 曹锦华,李华.多糖免疫作用的研究进展. 中国生化药物杂志,1999,20(2):104-105.
11. 王一飞,吕玉民主编.中药与肿瘤免疫研究.北京:中国医药科学出版社,1996:278-290.
12. 周世文,徐传福.多糖的免疫药理作用[J].中国生化药物志,1994,15(2):143.
13. 王蘅文主编.实验肿瘤学基础.北京.人民卫生出版社.1992.18-19.
14. 刘福英,王秀芳,冯旭.可移植性肿瘤动物模型的复制应用及问题.医学动物 治.2000,16(9):482-486.
15. 郭鹞主编.人类疾病的动物模型.北京.人民卫生出版社.1990:34.
16. 汪谦主编.现代医学实验方法.北京.人民卫生出版社.1998:1094.
17. 中华人民共和国药典委员会(一部)〔S〕.北京:化学工业出版社,2000,233.
18. 罗永东. 党参多糖提取工艺及制剂研究. 江西中医学院学报,1996 年增刊.11.
19. 熊元君,陈敏,李晓瑾,等.新疆党参多糖对 SOD、MDA 的影响. 新疆中医药 2000,18(3):13.
20. 陈敏,熊元君,李晓瑾,等.新疆党参和潞党参对小鼠脑 SOD、MDA 的影响. 中草药, 2000,31(4):280-281.
21. 张晓君,祝晨,胡黎.党参多糖对小鼠免疫和造血功能的影响. 中药新药与临床 药理,2003,14(3):174-175.
22. 黄泰康,李祥,陈建伟,等.明党参水煎液及多糖的药理研究.中成药, 994,16(7):31-33.
23. 李艳,鲁建江,孙萍,等.新疆党参多糖的提取及含量测定.新疆中医药,2001,19(3):9-10.
24. 田庚元,王晨,冯宇澄. 枸杞子糖蛋白的分离纯化、物化性质及糖肽键特征. 生物化学与生物物理学报,1995,27(2):201-205.
25. 夏尔宁,陈琼华.黑木耳多糖的分离纯化和鉴定.生物化学与生物物理学报,1988,20(6):614-618.
26. 方积年.多糖的分离纯化及其纯度鉴别与分子量测定.药理学通报,1984,19 (10):46-49.
27. W.preznic and R.H.F.Beck.New high-performance gel permeation chromatog raphic system for the determination of low-molecular-weight amyloses. J.Chromatogr,1987,387(5):467-472.
28. 国家药典委员会.中华人民共和国药典[S].2000年版二部.北京:化学工业出版 社,2000.附录ⅤH.
29. 李艳,鲁建江,孙萍,等. 新疆党参多糖的提取及含量测定. 新疆中医药, 2001,19(3):9-10.
30. 王莉,鲁建江,顾承志,等. 天花粉多糖的微波提取及含量测定. 药学实践杂 志,2001,19(3):54-55.
31. 马宝骊.免疫学技术 [M ].上海:上海医科大学出版社, 1984.188-190.
32. A.M.Staub. Removal of Proteins:Sevag Method,in Whistler RL(eds)Methods in Carbohydr. Chem, 1965,5:5-6.
33. 吴立军主编. 天然药物化学. 第四版:73-74.
34. Dubois M, Gilles KA, Hamilton JK, et al. Colorimetric method for determination of sugars and related substances. A Nal Chem , 1956,28(1):350-355.
35. Dubois M, Gilles KA,Hamilton JK, et al. A colorimetric method for the determination of sugars. Nature, 1951,168(4265):167.
36 . Michelsen KS.The roll of Toll—like receptors in bacteria—induced maturation of murine dendritic cells.J Biol Chem,2001,276(28):25680 —25686.
37. 韩锐主编.抗癌药物研究与实验技术.北京:北京医科大学中国协和医科 大学联合出版社,1997,67.
38. 徐淑云,卞如濂,陈修.药理实验方法学[M].第2版.北京:人民卫生出版社, 1992:l 423～I 440.
39. 李仪奎.中药药理实验方法学[M].上海:上海科学技术出版社,1991.172-175
40. 王慧铭,黄素霞,潘宏铭.香菇多糖对荷瘤小鼠免疫调节作用的研究.中华现代中西医杂志,2005,(3):22-25
41. 许华,杨晓明,杨锦南,等.脂麻花醇提物对 180 和 H22 小鼠的抗肿瘤作用研究, 中药材,2003,26(4):272-273
42. 何学斌,薛存宽,等.生膝多糖抗肿瘤作用的实验研究,肿瘤防治研究,2003,30(1):42-44.
43. 于善谦,免疫学导论[M].高等教育出版社,1999.
44 . 周永.多糖类抗肿瘤作用的研究进展,国外医学.卫生学册,2001,28(3):129-132
45. 杨建英.中西医结合治疗化疗中白细胞减少症[J].河南肿瘤学杂志,1999,8 (2) : 781
46. Ma G, Alexandar HR. Prevalence and pathophysiology of cancer cachexia. In :Bruera E ,Portenoy RK, eds : topics in palliative care , vol2[M] . New York :Oxford University Press ,1998. 912
47 . Bozzetti F , Migliavacca S , Scotti A et al . Impact of cancer ,type , site , stage and treatment on the nutritional status of patients[J].Ann Surg,1982,196 (2) :1702179.
48. 薛庆善 主编.提外培养的原理与技术.科学出版社.北京,2001:452-456
49. 万京华,章晓联,辛善禄.黄豆芽多糖对免疫抑制小鼠的实验研究.现代食品科技,2006,22(1):99-100
50. 姚金凤,吴 亮,吴希哲.海洋真菌多糖YCP对巨噬细胞免疫功能的影响.中国药科大学学报, 2004,35(6):573～57
51. 王晓京,丁桂风.范少光.几种阿片肽及ACTH对小鼠腹腔巨噬细胞吞噬功 能的作用[J].中国免疫学杂志,1987,3(4):211
1. Franz G. Polysaccharides in pharmacy:current applications and future concepts. Planta Med,1989,55(6):493-497.
2. Xu GM.PSP. International Symposium 1993.Fudan University Press:179
3. 田庚元.植物多糖的研究进展[J].中国中药杂志,1995,20(7):441-444.
4. 周建华. 国外对多糖的抗肿瘤作用研究简况. 国外医学 中医中药分册,1991,28(6):1-4.
5. 秦红,徐巧元. 香菇多糖与化疗联合应用治疗恶性胸水的疗效观察. 重庆医学,2000,29(5):463-464.
6. 李东霞,苏秀兰.多糖化合物抗肿瘤机制研究进展.中国新药杂志,2002,11(9):689-691
7. Kamei H,Hashimoto Y,Koide T,et al. Irect tumor growth suppressive effect of melanoidin extracted from immunomodulator-PSK[J]Cancer Biother Radiopharm, 1997,12(5):341～344.
8. 周静.近年来国内植物多糖活性研究进展.中草药,1994,25(1):40-44.
9. 唐小江,祝寿芬.山大黄及其多糖的急性毒性和抗突变作用试验.卫生毒理学 杂志,2000,14(1):43.
10. 宋剑秋,徐誉泰,张华坤,等.海带硫酸多糖对小鼠腹腔巨噬细胞的免疫调节作用. 中国免疫学杂志,2000,16(2):70.
11. 李绍平,季晖,李萍,等.冬虫夏草抗肿瘤作用研究进展.中草药,2001,32(4):373-375.
12. 王庆兰 , 张学仁 , 郭长强 , 等 . 枸杞子抗肿瘤作用研究综述 . 时珍国医国药,2001,11(6):559-560.
13. 康白,李广宙,张磊.香菇多糖对肿瘤浸润淋巴细胞体外扩增及抗瘤活性的影响. 中级医刊,2000,35(5):46-47.
14. 常雅平,杨贵贞.几种中药提取物诱生人干扰素.中国实验临床免疫学杂志, 1992,29(2):37-40.
15. Kim KH,Lee YS,Jung IS,et al. Acidic polysaccharide from Panax ginseng, ginsan, induces Th1 cell and macrophage cytokines and generates LAK cells in synergy with rIL-2.[J]Planta Med, 1998,64(2):110-115.
16.魏小龙,茹祥斌.低分子质量地黄多糖体外对 Lewis 肺癌细胞P53 基因表达 的影响.中国药理学通报,1998,14(3):245-248.
17.吕晓英,曾令福,李由,等.红毛五加多糖诱导胃癌细胞凋亡研究.中国新药杂志, 2000,9(3):166-167.
18.佟丽,黄添友,李及来,等.植物多糖对S180、K562 细胞增殖和唾液酸、磷酸、胆固醇含量的影响.中国中西医结合杂志,1994,14(8):482-484.
19.周永.多糖类抗肿瘤作用的研究进展.国外医学 卫生学分册,2001,28(3):129-132.
20.Nose M, Terawaki K, Oguri K,et al. Activation of macrophages by crude polysaccharide fractions obtained from shoots of Glycyrrhiza glabra and hairy roots of Glycyrrhiza uralensis in vitro.[J]Biol Pharm Bull,1998,21(10):1110-1112.
21. Zhang YW,Kiyohara H,Matsumoto T,et al. Fractionation and chemical properties ofimmunomodulating polysaccharides from roots of Dipsacus asperoides. Planta Med, 1997,63(3):393-399.
22. Shimizu N,Tomoda M,Kanari M,et al. An acidic polysaccharides having acitivity on the reticuloendothelial system from the root of Astragalus mongholicus.[J] Chem Pharm Bull, 1991,39(11):2969.
23 amaoka Y,Kawakita T,Kaneko M,etal. A polysaccharide fraction of Shosaikoto –To active in augmentation of natural killer activity by oral administration. [J]Biol Pharm Bull, 1995,18(6):846-849.
24. Liu F,Fung MC,Ooivec,et al. Induction in the mouse of gene expression of immunomodulating cytokines by Mushroom polysaccharide-protein complexes.[J] Life Science, 1996,58(21):1795～1803.
25. 谢学建,张俊慧,马爱华.商陆多糖研究进展. 时珍国医国药,1999,10(1):68-69.
26. Liu CP,Fang JN,Li XY,et al. Structural characterization and biological activities of SC4,an acidic polysaccharide from Salvia chinensis. Acta Pharmacol.Sin, 2002,23(2):162-166.
27. Hikno H, Ishiyama M, Suzuki Y, et al. Mechanisms of hypoglycemic activity of gandodern B: a glycan of ganoderma lucidum fruit bodies. [J]Planta Medica, 1989,55(2):423-428.
28. 徐梓辉,周世文,黄林清.薏苡仁多糖的分离提取及其降血糖作用的研究. 第三军医大学学报,2000,22(6):578-581.
29. 黄志江,季晖,李萍,等.人工虫草多糖降血糖作用极其机制研究.中国药科大学学报,2002,33(1):51-54.
30. Hikino H,Murakami M,Oshima Y.Isolation and hypoglycemic activity of oryzarans A,B,C,and D:glycans of Oryza sativa roots. Planta Med, 1986,(6):490-492.
31. Hikino H,Takahashi M,Oshima Y,et al. Related isolation and hypoglyccmic activity of oryzabrans A,B,C and D, glycans of Oryza sativabran. Planta Med, 1988,54(1):1-3.
32. Amornrut C,Toida T,Imanari T,et al.A new sulfated β-galactan from clams with anti-HIV activity. Carbohydr.Res, 1999,321(1-2):121-127.
33. Damonte E,Neytes J,Pujol CA,et al.Antiviral activity of a sulfated polysaccharides from the red seaweed Nothogenia fastigiata.Biochem.Pharnacol,1994,47(12): 2187-2192.
34. Mitsuya H,Looney DJ,Kuno S,et al.Dextran sulfate suppression of virues in the HIV family:inhibiyion of virion binding to CD4+cells.Science, 1988,240(4852): 646-649.
35. Baba M,Pauwels R,balzarini J,et al .Mechanism of inhibitory effect of dextan sulfate and heparin on replication of human immunodeficiency virus in vitro. Proc.Natl.Acda.Sci.USA, 1988,85(16):6132-6136.
36. Sano Y.Antiviral activity of chondroitin sulfate against infection by tobacco mosaic virus. Carbohydr.Poly, 1997,33(2):125-129.
37. 蒋岩,陈春英,裴丽健,等.硫酸酯化箬叶多糖抗 HIV-1 机制的初步研究.中华实验和临床病毒学杂志,2000,14(1):56-59.
38. 古元冬,史建勋,胡卓逸. 魔芋多糖的抗衰老作用.中草药,1999,30(2):127-128.
39. 陈智松,农志飞,吴志奎,等.枸杞多糖对衰老小鼠原癌基因表达的影响. 中草药,1998,28(5):356-359.
40. R.Dace,E.McBride,K.Brooks,et al.Comparison of the anticoagulant action of sulfated and phosphorylated polysaccharides. Thrombosis Research, 1997,87(1):113-121.
41. 杨彬. 糖类生物学:药物设计的新领域. 国外医药 药学分册. 1992,19(4):219-221.
42. 冯人田,王福清.低分子肝素的作用机制及其临床研究进展.中国生化药物杂志,1998,19(5):217-219.
43.陈建济,吴志强,王秀敏,等.灵芝多糖的药效学研究. 海峡医学,2000,12(1):51-55.
44. Hofer M,Pospisil M.Glucan as stimulator of hematoiesis in normal and gamma-irradiated mice.A survey of the authors’s results.Int J Immunopharmacol. 1997,19(9):607-609.
45. Qiu,Z,Mahiou B.Process for the preparation of immunomodulatory polysaccharides from aloe United States Patent,2000.Patent NO.6133440.
46. 刘曙晨,张慧娟,骆传环,等.猴头菇多糖的抗辐射作用实验研究.中华放射医学与防护杂志,1999,19(5):328-329.
47. 芮菁. 香菇多糖的药理作用和临床应用概况. 天津药学, 2002,12(1):35-37.
48. 陈建国. 茶多糖的提取及其药理作用的研究. 中草药,2000,31(7):附 6-附 7.
49. 汤军,钱华,黄琦,等.麦冬多糖平喘和抗过敏作用研究. 中国现代应用药学,1999(16):16-19.
50. 陈万生,韩军,李力,等.知母总多糖的抗炎作用.第二军医大学学报,1999,20(10):758-760.
51. Kim HS, Lee BM.. Inhibition of benzo[a]pyrene-DNA adduct formation by Aloe barbadensis Miller. [J]Carcinogenesis, 1997 ,18(4):771-776.
52 . Kim HS, Kacew S, Lee BM.In vitro chemopreventive effects of plant polysaccharides (Aloe barbadensis miller, Lentinus edodes, Ganoderma lucidum and Coriolus versicolor). [J]Carcinogenesis,1999,20(8):1637-1640.
2. 李春德译. 一种新的治疗药物的来源--糖类的研究[J]. 国外医学 药学分册,1994,21(6):353-356.
3. 李铁民,梁再赋. 甘草提取物及其衍生物的抗病毒研究现状[J].中草药,1994,25(12):655-658.
4. 张松,徐章.多糖类医药生物活性研究进展[J].中国生化药物志,1996,17(6):272-275.
5. 赵勇,张玲. 中药免疫调节研究进展[J]. 中草药,1994,25(11):603-606.
6. 丁媛媛. 多糖与抗病毒作用. 国外医学-微生物学分册,2001,24(1):18-20.
7. 李建恒,张杏红. 抗肿瘤中药多糖研究进展.中医药学报,1998,26(4):46-48.
8. Srubastava R,Kulshreshtha D.K. Biocide polysaccharides from plants. Phytochemistry, 1989,28(11):2877-2883.
9. 方积年.多糖研究的现状. 药学学报,1986,21(12):944-948.
10. 曹锦华,李华.多糖免疫作用的研究进展. 中国生化药物杂志,1999,20(2):104-105.
11. 王一飞,吕玉民主编.中药与肿瘤免疫研究.北京:中国医药科学出版社,1996:278-290.
12. 周世文,徐传福.多糖的免疫药理作用[J].中国生化药物志,1994,15(2):143.
13. 王蘅文主编.实验肿瘤学基础.北京.人民卫生出版社.1992.18-19.
14. 刘福英,王秀芳,冯旭.可移植性肿瘤动物模型的复制应用及问题.医学动物 治.2000,16(9):482-486.
15. 郭鹞主编.人类疾病的动物模型.北京.人民卫生出版社.1990:34.
16. 汪谦主编.现代医学实验方法.北京.人民卫生出版社.1998:1094.
17. 中华人民共和国药典委员会(一部)〔S〕.北京:化学工业出版社,2000,233.
18. 罗永东. 党参多糖提取工艺及制剂研究. 江西中医学院学报,1996 年增刊.11.
19. 熊元君,陈敏,李晓瑾,等.新疆党参多糖对 SOD、MDA 的影响. 新疆中医药 2000,18(3):13.
20. 陈敏,熊元君,李晓瑾,等.新疆党参和潞党参对小鼠脑 SOD、MDA 的影响. 中草药, 2000,31(4):280-281.
21. 张晓君,祝晨,胡黎.党参多糖对小鼠免疫和造血功能的影响. 中药新药与临床 药理,2003,14(3):174-175.
22. 黄泰康,李祥,陈建伟,等.明党参水煎液及多糖的药理研究.中成药, 994,16(7):31-33.
23. 李艳,鲁建江,孙萍,等.新疆党参多糖的提取及含量测定.新疆中医药,2001,19(3):9-10.
24. 田庚元,王晨,冯宇澄. 枸杞子糖蛋白的分离纯化、物化性质及糖肽键特征. 生物化学与生物物理学报,1995,27(2):201-205.
25. 夏尔宁,陈琼华.黑木耳多糖的分离纯化和鉴定.生物化学与生物物理学报,1988,20(6):614-618.
26. 方积年.多糖的分离纯化及其纯度鉴别与分子量测定.药理学通报,1984,19 (10):46-49.
27. W.preznic and R.H.F.Beck.New high-performance gel permeation chromatog raphic system for the determination of low-molecular-weight amyloses. J.Chromatogr,1987,387(5):467-472.
28. 国家药典委员会.中华人民共和国药典[S].2000年版二部.北京:化学工业出版 社,2000.附录ⅤH.
29. 李艳,鲁建江,孙萍,等. 新疆党参多糖的提取及含量测定. 新疆中医药, 2001,19(3):9-10.
30. 王莉,鲁建江,顾承志,等. 天花粉多糖的微波提取及含量测定. 药学实践杂 志,2001,19(3):54-55.
31. 马宝骊.免疫学技术 [M ].上海:上海医科大学出版社, 1984.188-190.
32. A.M.Staub. Removal of Proteins:Sevag Method,in Whistler RL(eds)Methods in Carbohydr. Chem, 1965,5:5-6.
33. 吴立军主编. 天然药物化学. 第四版:73-74.
34. Dubois M, Gilles KA, Hamilton JK, et al. Colorimetric method for determination of sugars and related substances. A Nal Chem , 1956,28(1):350-355.
35. Dubois M, Gilles KA,Hamilton JK, et al. A colorimetric method for the determination of sugars. Nature, 1951,168(4265):167.
36 . Michelsen KS.The roll of Toll—like receptors in bacteria—induced maturation of murine dendritic cells.J Biol Chem,2001,276(28):25680 —25686.
37. 韩锐主编.抗癌药物研究与实验技术.北京:北京医科大学中国协和医科 大学联合出版社,1997,67.
38. 徐淑云,卞如濂,陈修.药理实验方法学[M].第2版.北京:人民卫生出版社, 1992:l 423～I 440.
39. 李仪奎.中药药理实验方法学[M].上海:上海科学技术出版社,1991.172-175
40. 王慧铭,黄素霞,潘宏铭.香菇多糖对荷瘤小鼠免疫调节作用的研究.中华现代中西医杂志,2005,(3):22-25
41. 许华,杨晓明,杨锦南,等.脂麻花醇提物对 180 和 H22 小鼠的抗肿瘤作用研究, 中药材,2003,26(4):272-273
42. 何学斌,薛存宽,等.生膝多糖抗肿瘤作用的实验研究,肿瘤防治研究,2003,30(1):42-44.
43. 于善谦,免疫学导论[M].高等教育出版社,1999.
44 . 周永.多糖类抗肿瘤作用的研究进展,国外医学.卫生学册,2001,28(3):129-132
45. 杨建英.中西医结合治疗化疗中白细胞减少症[J].河南肿瘤学杂志,1999,8 (2) : 781
46. Ma G, Alexandar HR. Prevalence and pathophysiology of cancer cachexia. In :Bruera E ,Portenoy RK, eds : topics in palliative care , vol2[M] . New York :Oxford University Press ,1998. 912
47 . Bozzetti F , Migliavacca S , Scotti A et al . Impact of cancer ,type , site , stage and treatment on the nutritional status of patients[J].Ann Surg,1982,196 (2) :1702179.
48. 薛庆善 主编.提外培养的原理与技术.科学出版社.北京,2001:452-456
49. 万京华,章晓联,辛善禄.黄豆芽多糖对免疫抑制小鼠的实验研究.现代食品科技,2006,22(1):99-100
50. 姚金凤,吴 亮,吴希哲.海洋真菌多糖YCP对巨噬细胞免疫功能的影响.中国药科大学学报, 2004,35(6):573～57
51. 王晓京,丁桂风.范少光.几种阿片肽及ACTH对小鼠腹腔巨噬细胞吞噬功 能的作用[J].中国免疫学杂志,1987,3(4):211
1. Franz G. Polysaccharides in pharmacy:current applications and future concepts. Planta Med,1989,55(6):493-497.
2. Xu GM.PSP. International Symposium 1993.Fudan University Press:179
3. 田庚元.植物多糖的研究进展[J].中国中药杂志,1995,20(7):441-444.
4. 周建华. 国外对多糖的抗肿瘤作用研究简况. 国外医学 中医中药分册,1991,28(6):1-4.
5. 秦红,徐巧元. 香菇多糖与化疗联合应用治疗恶性胸水的疗效观察. 重庆医学,2000,29(5):463-464.
6. 李东霞,苏秀兰.多糖化合物抗肿瘤机制研究进展.中国新药杂志,2002,11(9):689-691
7. Kamei H,Hashimoto Y,Koide T,et al. Irect tumor growth suppressive effect of melanoidin extracted from immunomodulator-PSK[J]Cancer Biother Radiopharm, 1997,12(5):341～344.
8. 周静.近年来国内植物多糖活性研究进展.中草药,1994,25(1):40-44.
9. 唐小江,祝寿芬.山大黄及其多糖的急性毒性和抗突变作用试验.卫生毒理学 杂志,2000,14(1):43.
10. 宋剑秋,徐誉泰,张华坤,等.海带硫酸多糖对小鼠腹腔巨噬细胞的免疫调节作用. 中国免疫学杂志,2000,16(2):70.
11. 李绍平,季晖,李萍,等.冬虫夏草抗肿瘤作用研究进展.中草药,2001,32(4):373-375.
12. 王庆兰 , 张学仁 , 郭长强 , 等 . 枸杞子抗肿瘤作用研究综述 . 时珍国医国药,2001,11(6):559-560.
13. 康白,李广宙,张磊.香菇多糖对肿瘤浸润淋巴细胞体外扩增及抗瘤活性的影响. 中级医刊,2000,35(5):46-47.
14. 常雅平,杨贵贞.几种中药提取物诱生人干扰素.中国实验临床免疫学杂志, 1992,29(2):37-40.
15. Kim KH,Lee YS,Jung IS,et al. Acidic polysaccharide from Panax ginseng, ginsan, induces Th1 cell and macrophage cytokines and generates LAK cells in synergy with rIL-2.[J]Planta Med, 1998,64(2):110-115.
16.魏小龙,茹祥斌.低分子质量地黄多糖体外对 Lewis 肺癌细胞P53 基因表达 的影响.中国药理学通报,1998,14(3):245-248.
17.吕晓英,曾令福,李由,等.红毛五加多糖诱导胃癌细胞凋亡研究.中国新药杂志, 2000,9(3):166-167.
18.佟丽,黄添友,李及来,等.植物多糖对S180、K562 细胞增殖和唾液酸、磷酸、胆固醇含量的影响.中国中西医结合杂志,1994,14(8):482-484.
19.周永.多糖类抗肿瘤作用的研究进展.国外医学 卫生学分册,2001,28(3):129-132.
20.Nose M, Terawaki K, Oguri K,et al. Activation of macrophages by crude polysaccharide fractions obtained from shoots of Glycyrrhiza glabra and hairy roots of Glycyrrhiza uralensis in vitro.[J]Biol Pharm Bull,1998,21(10):1110-1112.
21. Zhang YW,Kiyohara H,Matsumoto T,et al. Fractionation and chemical properties ofimmunomodulating polysaccharides from roots of Dipsacus asperoides. Planta Med, 1997,63(3):393-399.
22. Shimizu N,Tomoda M,Kanari M,et al. An acidic polysaccharides having acitivity on the reticuloendothelial system from the root of Astragalus mongholicus.[J] Chem Pharm Bull, 1991,39(11):2969.
23 amaoka Y,Kawakita T,Kaneko M,etal. A polysaccharide fraction of Shosaikoto –To active in augmentation of natural killer activity by oral administration. [J]Biol Pharm Bull, 1995,18(6):846-849.
24. Liu F,Fung MC,Ooivec,et al. Induction in the mouse of gene expression of immunomodulating cytokines by Mushroom polysaccharide-protein complexes.[J] Life Science, 1996,58(21):1795～1803.
25. 谢学建,张俊慧,马爱华.商陆多糖研究进展. 时珍国医国药,1999,10(1):68-69.
26. Liu CP,Fang JN,Li XY,et al. Structural characterization and biological activities of SC4,an acidic polysaccharide from Salvia chinensis. Acta Pharmacol.Sin, 2002,23(2):162-166.
27. Hikno H, Ishiyama M, Suzuki Y, et al. Mechanisms of hypoglycemic activity of gandodern B: a glycan of ganoderma lucidum fruit bodies. [J]Planta Medica, 1989,55(2):423-428.
28. 徐梓辉,周世文,黄林清.薏苡仁多糖的分离提取及其降血糖作用的研究. 第三军医大学学报,2000,22(6):578-581.
29. 黄志江,季晖,李萍,等.人工虫草多糖降血糖作用极其机制研究.中国药科大学学报,2002,33(1):51-54.
30. Hikino H,Murakami M,Oshima Y.Isolation and hypoglycemic activity of oryzarans A,B,C,and D:glycans of Oryza sativa roots. Planta Med, 1986,(6):490-492.
31. Hikino H,Takahashi M,Oshima Y,et al. Related isolation and hypoglyccmic activity of oryzabrans A,B,C and D, glycans of Oryza sativabran. Planta Med, 1988,54(1):1-3.
32. Amornrut C,Toida T,Imanari T,et al.A new sulfated β-galactan from clams with anti-HIV activity. Carbohydr.Res, 1999,321(1-2):121-127.
33. Damonte E,Neytes J,Pujol CA,et al.Antiviral activity of a sulfated polysaccharides from the red seaweed Nothogenia fastigiata.Biochem.Pharnacol,1994,47(12): 2187-2192.
34. Mitsuya H,Looney DJ,Kuno S,et al.Dextran sulfate suppression of virues in the HIV family:inhibiyion of virion binding to CD4+cells.Science, 1988,240(4852): 646-649.
35. Baba M,Pauwels R,balzarini J,et al .Mechanism of inhibitory effect of dextan sulfate and heparin on replication of human immunodeficiency virus in vitro. Proc.Natl.Acda.Sci.USA, 1988,85(16):6132-6136.
36. Sano Y.Antiviral activity of chondroitin sulfate against infection by tobacco mosaic virus. Carbohydr.Poly, 1997,33(2):125-129.
37. 蒋岩,陈春英,裴丽健,等.硫酸酯化箬叶多糖抗 HIV-1 机制的初步研究.中华实验和临床病毒学杂志,2000,14(1):56-59.
38. 古元冬,史建勋,胡卓逸. 魔芋多糖的抗衰老作用.中草药,1999,30(2):127-128.
39. 陈智松,农志飞,吴志奎,等.枸杞多糖对衰老小鼠原癌基因表达的影响. 中草药,1998,28(5):356-359.
40. R.Dace,E.McBride,K.Brooks,et al.Comparison of the anticoagulant action of sulfated and phosphorylated polysaccharides. Thrombosis Research, 1997,87(1):113-121.
41. 杨彬. 糖类生物学:药物设计的新领域. 国外医药 药学分册. 1992,19(4):219-221.
42. 冯人田,王福清.低分子肝素的作用机制及其临床研究进展.中国生化药物杂志,1998,19(5):217-219.
43.陈建济,吴志强,王秀敏,等.灵芝多糖的药效学研究. 海峡医学,2000,12(1):51-55.
44. Hofer M,Pospisil M.Glucan as stimulator of hematoiesis in normal and gamma-irradiated mice.A survey of the authors’s results.Int J Immunopharmacol. 1997,19(9):607-609.
45. Qiu,Z,Mahiou B.Process for the preparation of immunomodulatory polysaccharides from aloe United States Patent,2000.Patent NO.6133440.
46. 刘曙晨,张慧娟,骆传环,等.猴头菇多糖的抗辐射作用实验研究.中华放射医学与防护杂志,1999,19(5):328-329.
47. 芮菁. 香菇多糖的药理作用和临床应用概况. 天津药学, 2002,12(1):35-37.
48. 陈建国. 茶多糖的提取及其药理作用的研究. 中草药,2000,31(7):附 6-附 7.
49. 汤军,钱华,黄琦,等.麦冬多糖平喘和抗过敏作用研究. 中国现代应用药学,1999(16):16-19.
50. 陈万生,韩军,李力,等.知母总多糖的抗炎作用.第二军医大学学报,1999,20(10):758-760.
51. Kim HS, Lee BM.. Inhibition of benzo[a]pyrene-DNA adduct formation by Aloe barbadensis Miller. [J]Carcinogenesis, 1997 ,18(4):771-776.
52 . Kim HS, Kacew S, Lee BM.In vitro chemopreventive effects of plant polysaccharides (Aloe barbadensis miller, Lentinus edodes, Ganoderma lucidum and Coriolus versicolor). [J]Carcinogenesis,1999,20(8):1637-1640.