海藻硒多糖的合成及生物活性研究
|
摘要
采用直接水提法对琼枝麒麟菜中的多糖进行提取，乙醇沉淀得麒麟菜多糖，红外光谱表明其主要为卡拉胶。在硝酸-氯化钡或硝酸催化作用下，利用亚硒酸钠对麒麟菜多糖进行分子修饰，合成了硒化麒麟菜多糖，透析除去过量的亚硒酸钠，抗坏血酸检测，约24小时不显红色时停止透析，较好的反应条件是硝酸体积浓度为0.5％，70℃反应10小时，透析24小时后，ICP测得此时硒含量为0.413mg／g，多糖回收率为0.73％。利用红外光谱详细分析了不同反应条件对合成硒多糖结构的影响，透析后硒多糖在930和850cm~(-1)的吸收峰强度减弱，这是因为硝酸存在下多糖部分降解成为小分子寡糖，并能在透析时被除去。对比不同反应条件下和不同透析时间得到硒多糖中的硒含量，亚硒酸根主要与小分子寡糖结合，硒多糖的核磁共振光谱也证实了硒多糖与多糖在结构上的差别。观察到硝酸-氯化钡或硝酸催化合成的硒多糖的效果相差不大，因此可以改进文献方法只用硝酸催化，此外还通过抗坏血酸还原过量亚硒酸钠，得到硒多糖分散的纳米硒，简化了提纯硒多糖的步骤。
采用四甲基偶氮唑盐比色法(MTT法)研究了硒多糖和硒多糖分散的纳米硒对人喉上皮细胞癌HEp-2、人白血病细胞K562及人肺癌细胞H460细胞株的抑制作用。硒多糖具有良好的抗HEp-2细胞癌活性，且抑制作用大于麒麟菜多糖和亚硒酸钠，硒多糖对K562和H460细胞癌的抑制作用大于麒麟菜多糖，但同等剂量条件下的病毒抑制率低于亚硒酸钠，各硒化麒麟菜多糖的抗癌活性具有浓度依赖性特点。硒多糖分散的纳米硒在低剂量下具有优良的抗HEp-2病毒活性，其IC_(50)为硒化麒麟菜多糖的1／291，可望开发成抗肿瘤药物。
本文还研究了等摩尔SDS／非离子表面活性剂反胶束中纳米硒的合成，探究了不同表面活性剂复配后对纳米硒形貌的影响。
Sulfated polysaccharide was extracted from Eucheuma gelatinae with distilled water, precipitated with ethanol. FT IR spectrum of the polysaccharide indicated that it is κ-carrageenan. Seleno-polysaccharides were prepared by employing sodium selenite to modify the polysaccharide under the catalysis of HNO_3-BaCl_2 or HNO_3. Dialysis was applied to remove the excess sodium selenite. It was about 24hrs to stop dialysis when the red detected by ascorbic acid stopped to appear. The favor reaction conditions were at 70℃ for 10 hours in the presence of 0.5% volume concentration of nitric acid. The selenic content determined by ICP of seleno-polysaccharide obtained under such condition was 0.413mg/g, while the yield of seleno-polysaccharide was 0.73 %. The absorption bands of seleno-polysaccharide at 930 and 850 cm~(-1) became weak after dialyzing. The reason is part of the macromolecular polysaccharide can be decomposed to form oligosaccharide which can be remove by dialysis in the presence of nitric acid. Selenite is bonded mainly to the oligosaccharide by comparing selenium contents in seleno-polysaccharides obtained under different reacting conditions and after dialyzing for different time. ~(13)C-NMR confirmed the structural difference between seleno-polysaccharide and polysaccharide. The effects catalyzed by HNO_3-BaCl_2 or HNO_3 on seleno-polysaccharide are similar. Thus, nitric acid can only be used as catalyst by improving the reported method. Excess selenite in the seleno-polysaccharide catalyzed by nitric acid was reduced by ascorbic acid, and nano-selenium dispersed by seleno-polysaccharide was prepared.
The inhibiting effects of seleno-polysaccharide and nano-selenium dispersed by seleno-polysaccharide on HEp-2, K562, and H460 were studied by MTT method. The results showed that seleno-polysaccharide significantly inhibited the proliferation of HEp-2 cells in vitro. Compared with natural polysaccharide or sodium selenite containing the same amount of Se, the seleno-polysaccharide was more effective. Seleno-polysaccharide was more effective on K562 and H460 cells than natural polysaccharide, but less effective than sodium selenite in the same amount. The inhibition relies on the concentration of the seleno-polysaccharide. Nano-selenium dispersed by seleno-polysaccharide revealed
采用四甲基偶氮唑盐比色法(MTT法)研究了硒多糖和硒多糖分散的纳米硒对人喉上皮细胞癌HEp-2、人白血病细胞K562及人肺癌细胞H460细胞株的抑制作用。硒多糖具有良好的抗HEp-2细胞癌活性，且抑制作用大于麒麟菜多糖和亚硒酸钠，硒多糖对K562和H460细胞癌的抑制作用大于麒麟菜多糖，但同等剂量条件下的病毒抑制率低于亚硒酸钠，各硒化麒麟菜多糖的抗癌活性具有浓度依赖性特点。硒多糖分散的纳米硒在低剂量下具有优良的抗HEp-2病毒活性，其IC_(50)为硒化麒麟菜多糖的1／291，可望开发成抗肿瘤药物。
本文还研究了等摩尔SDS／非离子表面活性剂反胶束中纳米硒的合成，探究了不同表面活性剂复配后对纳米硒形貌的影响。
Sulfated polysaccharide was extracted from Eucheuma gelatinae with distilled water, precipitated with ethanol. FT IR spectrum of the polysaccharide indicated that it is κ-carrageenan. Seleno-polysaccharides were prepared by employing sodium selenite to modify the polysaccharide under the catalysis of HNO_3-BaCl_2 or HNO_3. Dialysis was applied to remove the excess sodium selenite. It was about 24hrs to stop dialysis when the red detected by ascorbic acid stopped to appear. The favor reaction conditions were at 70℃ for 10 hours in the presence of 0.5% volume concentration of nitric acid. The selenic content determined by ICP of seleno-polysaccharide obtained under such condition was 0.413mg/g, while the yield of seleno-polysaccharide was 0.73 %. The absorption bands of seleno-polysaccharide at 930 and 850 cm~(-1) became weak after dialyzing. The reason is part of the macromolecular polysaccharide can be decomposed to form oligosaccharide which can be remove by dialysis in the presence of nitric acid. Selenite is bonded mainly to the oligosaccharide by comparing selenium contents in seleno-polysaccharides obtained under different reacting conditions and after dialyzing for different time. ~(13)C-NMR confirmed the structural difference between seleno-polysaccharide and polysaccharide. The effects catalyzed by HNO_3-BaCl_2 or HNO_3 on seleno-polysaccharide are similar. Thus, nitric acid can only be used as catalyst by improving the reported method. Excess selenite in the seleno-polysaccharide catalyzed by nitric acid was reduced by ascorbic acid, and nano-selenium dispersed by seleno-polysaccharide was prepared.
The inhibiting effects of seleno-polysaccharide and nano-selenium dispersed by seleno-polysaccharide on HEp-2, K562, and H460 were studied by MTT method. The results showed that seleno-polysaccharide significantly inhibited the proliferation of HEp-2 cells in vitro. Compared with natural polysaccharide or sodium selenite containing the same amount of Se, the seleno-polysaccharide was more effective. Seleno-polysaccharide was more effective on K562 and H460 cells than natural polysaccharide, but less effective than sodium selenite in the same amount. The inhibition relies on the concentration of the seleno-polysaccharide. Nano-selenium dispersed by seleno-polysaccharide revealed
引文
[1] 许实波，海洋生理活性物质的研究及发展趋势[J]，生物工程进展，1990，16(6)：25-30
[2] 王长云，耿美玉，管华诗。海洋药物研究进展与发展趋势[J]，中国新药杂志，2005，14(3)，278-282
[3] 盛建春，杨方美，胡秋辉，海藻多糖生物活性研究[J]，食品科学，2005，26(3)，262-264
[4] 张翼，李晓明，王斌贵，海藻生物活性物质研究的回顾与展望[J]，世界科技研究与发展，2005，27(05)：56-63
[5] 盛建春，杨方美，胡秋辉，海藻多糖生物活性研究[J]，食品科学，2005，26(3)，262-264
[6] 王长云，管华诗，多糖抗病毒作用研究进展Ⅰ．多糖抗病毒作用[J]，生物工程进展，2000，20(1)：17-20
[7] Woo E R, Kim W S, Kim Y S, Virus-cell fusion inhibitory activity for the polysaccharides from various Korean edible clams[J], Arch. Pharm. Res., 2001, 24(6): 514—517
[8] Huheihel M, Ishanu V, Tal J, et al, Activity of Porphyridium sp. Polysaccharide against herpes simplex viruses in vitro and in vivo[J], Biochem. Biophys. Methods, 2002, 50:189-200
[9] 辛现良，耿美玉，李桂玲，管华诗，李泽琳，海洋硫酸多糖911体外对HIV-1的作用研究[J]，中国海洋药物，2000，(3)：8-10
[10] Fumihiko T, Rieko N, Tomoko K, et al, Lentinan aufrnent skin reation induced by bradykinin: its correlation with vascular dilatation and hemorrhage responses and antitumor activities[J], Internat immunopharmacol, 1995,17(6):465-474
[11] Kim K H, Lee Y S, Jung I S, et al, Acidic polysaccharide from Panax ginseng, ginsan, induces Thl cell and macrophage cytokines and generates LAK cells in synergy with rIL-2[J], Planta. Med., 1998, 64(2): 110-115
[12] Koyanagi S, Tanigawa N, Nakagawa H, et al, Oversulfation of fucoidan enhances its anti-angiogenic and antitumor activities[J], Biochemi Pharmacol, 2003, 65:173-179
[13] Katsuhide M, Shin Y, Yuji K, et al, Activity of microbial surface polysaccharides in inhibition of cancer cell adhesion[J], Kagaku Kogaku, 1996, 60(11): 932
[14] Sogawa K, Sumida T, Hamakawa H, et al, Inhibitory effect of a marine microalgal polysaccharide on thetelomerase activity in K562 cells[J], Research Communications in Molecular Pathology and Pharmacology, 1998, 99(3): 259-265
[15] Zhang M, Zhang L N, Cheung Keung P C, et al, Molecular weight and anti-tumor activity of the water-soluble polysaccharides isolated by hot water and ultrasonic treatment from the sclerotia and mycelia of Pleurotus tuber-regium[J], Carbohydrate Polymers, 2004, 56: 123-128.
[16] 师然新，徐祖洪，李恩智，降解的角叉菜多糖的抗肿瘤活性[J]，海洋与湖沼，2000，31(6)：45—48
[17] 季宇彬，张琪，孙红，杨卫东，夏月，羊栖菜多糖对P_(388)小鼠红细胞免疫促进作用的机制研究[J]，中国海洋药物，1998，(2)：14—18
[18] 何雅军，吴谦，文蛤提取物对小鼠DTH反应的双向免疫调节作用[J]，中国海洋药物，1995，(3)：20—22
[19] 杨晓林，孙菊云，许汉年，刘晓慧，张翼伸，张绍伦，褐藻糖胶的免疫调节作用[J]，中国海洋药物，1995，(3)：9—12
[20] 刘茜，焦庆才，刘志礼，螺旋藻多糖及其药理作用的研究进展[J]，中国海洋药物，1998，(1)：48—50
[21] Hu J F, Geng M Y, Zhang J T, et al. An in vitro study of the structure-activity relationships of sulfated polysaccharide from brown algae to its antioxidant effect[J]. J. Asian Nat. Prod. Res., 2001, 3(4): 353—358
[22] Yoon S J, Yu M A, Pyun Y R, et al, The nontoxic mushroom auricularia auricular contains a polysaccharide with anticoagulant activity mediated by antithrombin[J], Thrombosis Research, 2003,(112): 151-158
[23] 刘成梅，付桂明，涂宗财，万茵，百合多糖降血糖功能研究[J]，食品科学，2002，23(6)：113-114
[24] Dawes C J, Stanley N F, Moon R E, Physiological and biochemical studies on the L-carrageenan producing red algae Eucheuma uncinatum Setchell and Gardner from the gulf of California[J], Bot. Mar., 1977, 20(7): 437-442
[25] 张其标，耳突麒麟菜卡拉胶提取工艺条件的优化选择[J]，食品科学，1999，(4)：34-36
[26] 韩国华，李海霞，吴杨桦，菲律宾刺生麒麟菜提取：ι-卡拉胶的工艺探讨[J]，食品与机械，2001，(3)：31-32
[27] 汤毅珊，赵谋明，高孔荣，麒麟菜卡拉胶提取新工艺的探讨[J]，食品工业科技，1997，(2)：30-34
[28] 林宇野，杨虹，酶法提取多糖的研究[J]，食品与发酵工业，1995，(1)：13-17
[29] 康琰琰，王一飞，朱良，门晓缓，裙带菜茎中硫酸多糖的分离纯化及分析鉴定[J]，中药材，2005，28(9)：769-771
[30] 王兆梅，李琳，郭祀远，蔡妙颜，活性多糖构效关系研究评述[J]，现代化工，2002，22(8)：18-21
[32] 吴立根，毛文君，衍生化多糖的生物活性研究进展[J]，海洋科学，2002，26(5)：23-25
[33] Dace R, Comparison of the anticoagulant action of sulfated and phosphorylated olysaccharides[J], Thrombosts Research,1997, 87(1): 113-121
[34] 邓成华，杨祥良，王雁，顾小曼，硫酸酯化及羧甲基化虎奶多糖的抗病毒作用[J]，中国地方病学杂志，2000，19(4)：251-253
[35] 孟春，郭养浩，石贤爱，陈剑锋，李锋，海藻多糖生物活性及分子修饰[J]，中国生物工程杂志，2004，24(3)：35-39
[36] 刘占峰，孙汉文，多糖的化学修饰研究进展[J]，河北大学学报(自然科学版)2005，25(1)：104-108
[37] 赖萍，林跃鑫，天然多糖分子修饰研究进展[J]，生命的化学，2003，23(3)：183-187
[38] Uryu T, Ikushima N,Katsuraya K, etal. Sulfated alkyl oligoaacharides with potent inhibitory effects on HIV infection.Biochemical Pharmacol, 1992, 43:2385-2392
[39] Tapiero H, Townsend D M, Tew K D, The antioxidant role of selenium and seleno-compounds[J], Biomedicine & Pharmacotherapy, 2003,(57): 134-144
[40] Gladshev V N, Khangulov S V, Stadtman T C, Properties of the selenium and molybdenum-containing nicotinic acid hydroxylase from clostridium barkeri[J], Biochemistry, 1996,(35): 1212-1233
[41] 刘建林，何新乡，陈宏，硒酸酯多糖的特性及其在食品工业中的应用[J]，中国食品添加剂，2003，(5)：80-82
[42] 许峰，张瞳，周波，贾乐，食用蕈菌硒多糖研究进展[J]，生物技术，2006，16(1)：86-88
[43] 戴伟娟，仲伟法，索金良，林志彬，4-硒硫酸酯多糖对人肝癌细胞Bel／7402DNA RNA及蛋白质合成的影响[J]，济宁医学院学报，2001，24(1)：18-19
[44] 陈展寰，等．无机化学新论选读[M]，西安：陕西人民出版社，1998
[45] 张新宇，王番，绿色巴夫藻硒多糖的提取、分离与纯化[J]，海洋与湖沼，2000，31(6)：643-645
[46] Yoshida M, Fukunaga K, Tsuchita H, et al. Anevaluation of the bio-availability of selenium in high-selenium yeast[J], J-Nutr-Sci-Vitaminol-Tokyo, 1999, 45(1): 119-128
[47] Spolar M R, Schaffer E M, Beelman R B, et al, Selenium-enriched Agaricus bisporus mushrooms suppress 7,12-dimethlybenz[a] anthracene bioactivation in mammary tissue[J], Cancer Lett., 1999, 138(122): 145～150
[48] 尚德静，王孝敏，许永廷，田星，安悦，SeGLP-1的分离纯化及硒活性结构分析[J]，食用菌学报，2002，9(1)：22-27
[49] 郑文杰，贺鸿志，黄峙，杨芳，螺旋藻富集和转化硒研究进展[J]，中国生物工程杂志，2003，23(1)：57-60
[50] 唐家骏，芮海风，硒化卡拉胶的制法，专利CN 1014522B
[51] 龚晓钟，欧阳政，硒化黄芪多糖制备条件及其结构的研究[J]，天然产物研究与开发，1997，10(2)：26-32
[52] 龚晓钟，欧阳政，硒化葡聚糖结构测定及药效研究[J]，深圳大学学报(理工版)，1998，15(1)：70-74
[53] 李光友，缪锦来，刘发义，硒多糖化合物及其制备方法，专利CN 1121414C
[54] 崔乔，尚德静，邹霞，硒多糖的研究进展[J]，中国生化药物杂志，2003，24(3)，155-157
[55] 尚德静，崔乔，高林，灵芝硒多糖SeGLP-1抑制小鼠肝腹水癌作用的研究[J]，食用菌学报，2001，3：34-38
[56] Combs G F, Garbisu C, Lee B C, et al, Bioavailability of selenium accumulated by selenite-reducing bacteria[J], Biological Trace Element Research, 1996, 52(3), 209-225
[57] Nuttall K L, Elemental selenium and glutathione reductase[J], Medical hypotheses, 1985, 16(2): 155-158
[58] 高学云，张劲松，张立德，朱茂祥，纳米红色元素硒对小鼠的免疫功能的调节作用[J]，中国公共卫生，2000，16(5)：421-422
[59] 张劲松，高学云，张立德，朱茂祥，杨陟华，蛋白质分散的纳米红色元素硒的延缓衰老作用[J]，营养学报，2000，22(3)：219-222
[60] Gao X Y, Zhang J S, Zhang L D, Hollow sphere selenium nanoparticles: their in-vitro anti hydroxyl radical effect[J], Advanced Materials, 2002, 14(4): 290-293
[61] Huang B, Zhang J S, Hou J W, Chen C, Free radical scavenging efficiency of Nano-Se in vitro[J], Free Radical Biology & Medicine, 2003, 35(7): 805-813
[62] Zhang JinSong, Gao XueYun, Zhang LiDe, Bao YongPing, Biological effects of a nano red elemental selenium[J], BioFactors, 2001, 15(1):27-38
[63] Zhang J S, Wang H L, Bao Y P, Zhang L D, Nano red elemental selenium has no size effect in the induction of seleno-enzymes in both cultured cells and mice[J], Life Sciences, 2004, 75(2): 237-244
[64] Johnson J A, Saboungi M L, Thiyagarajan P, Csencsits R, Meisel D, Selenium Nanoparticles: A Small-Angle Neutron Scattering Study[J], Journal of Physical Chemistry B, 1999, 103(1): 59-63
[65] Abdelouas A, Gong W L, Lutze W, et al, Using cytochrome C3 to make selenium nanowires[J], Chem. Mater., 2000,12:1510～1512
[66] Gates B, Yin Y D, Xia Y N, et al, A solution-phase approach to the synthesis of uniform nanowires of crystalline selenium with lateral dimensional in the range of 10-30nm[J], J. Am. Chem. Soc., 2000, 122:12582～12583
[67] Mees D R, Pysto W, Tarcha P J, Formation of selenium Colloids Using Sodium Ascorbate as the Reducing Agent[J], Journal of Colloid and Interface Science, 1995, 170:254-260
[68] Gao X Y, Zhang J S, Zhang L D, Hollow Sphere Selenium Nanoparticles: There In-vitro Anti Hydroxyl Radical Effect[J], Adv. Mater., 2002, 14(4): 290-293
[69] 张胜义，张娟，刘明珠，朱俊杰，陈洪渊，PVP存在下液液界面生长法制备硒纳米线[J]，无机化学学报，2005，21(1)：137-141
[70] Gates B, Mayers B, Grossman A, et al, A Sonochemical Approach to the Synthesis of Crystalline Selenium Nanowires in Solutions and on Solid Supports[J],Adv. Mater., 2000, 14(23): 1749-1752
[71] Gates B, Mayers B, Cattle B, et al, Synthesis and Characterization of Uniform Nanowires of Trigonal Selenium Whose Diameters Could Be Controlled in the Range of 10-800nm[J]. Adv. Funct. Mater., 2002, 12(3): 219-227
[72] Bunker C E, Harruff B A, Pathak P, et al, Formation of cadmium sulfide nanoparticles in reverse micelles: Extreme sensitivity to preparation procedure[J], Langmuir,2004,20(13): 5642-5644
[73] Palla B J, Shah D O, Stabilization of high ionic strength slurries using surfactant mixtures: molecular factors that determine optimal stability[J], Journal of Colloid and Interface Science, 2002, 256:143-152
[74] Tian B Z, Liu X Y, Zhang Z D, Tu B, Zhao D Y, Syntheses of High-Quality Mesoporous Materials Directed by Blends of Nonionic Amphiphiles under Nonaqueous Conditions[J], Journal of Solid State Chemistry, 2002, 167(2): 324-329
[75] Wang Y D, Ma C L, Sun X D, Li H D, Synthesis and characterization of ordered hexagonal and cubic mesoporous tin oxides via mixed-surfactant templates route[J], Journal of Colloid and Interface Science, 2005, 296:627-631
[76] Shen Q, Wei H, Wang L C, et al, Crystallization and aggregation behaviors of calcium carbonate in the presence of poly(vinylpyrrolidone) and sodium dodecyl sulfate[J], Journal of Physical Chemistry B, 2005, 109(39): 18342-18347
[77] Yu J G, Zhao X F, Cheng B, et al, Controlled synthesis of calcium carbonate in a mixed aqueous solution of PSMA and CTAB[J], Journal of Solid State Chemistry, 2005, 178(3): 861-867
[78] Kang S H, Hirasawa I, Kim W S, et al, Morphological control of calcium carbonate crystallized in reverse micelle system with anionic surfactants SDS and AOT[J], Journal of Colloid and Interface Science, 2005, 288(2): 496-502
[79] Somasundaran P, Changes in micelle compositions and monomer concentrations in mixed surfactant solutions[J], Langmuir, 1996, 12:5790-5795
[80] Md Emdadul Haque, Akhil Ranjandas, Satya priya moulik J, Mixed micelles of sodium deoxycholate and polyoxyethylene sorbitan monooleate(Tween80)[J], Journal of Colloid and Interface Science, 1999, 217:1-7
[81] 高景镰，麒麟菜的开发利用，中国野生植物[J]，1991，(2)：31-33
[82] 吴风山，吴涤，光度法测定黑木耳中微量硒[J]，光谱实验室，2004，21(4)：719-721
[83] 刘庆堂，李锐增，李坚，紫外分光光度法测定龙胆中微量元素硒[J]，广东药学，2001，11(6)30-32
[84] 高静娴，谷学新，赵晓倩，微孔滤膜富集分光光度法测定痕量硒[J]，首都师范大学学报，1996，17(1)：60-64
[85] 杨金辉，杨江柳，胡鄂明，邓喆，柳建祥，电感耦合等离子体发射光谱法(ICP-AES)测定铀矿石中微量钪、硒、铼[J]，南华大学学报(理工版)，2004，18(4)：83-85
[86] 梁智渊，岑颖洲，叶绍明，伍秋明，琼枝和异枝麒麟菜中硫酸酯基多糖不同提取方法的化学分析比较[J]，暨南大学学报，2005，26(3)：380-385
[87] Prado-Fernandez J, Rodriguez-Vazquez J A, Tojo E, Andrade J M, Quantitation of κ-, ι-and λ-carrageenans by mid-infrared spectroscopy and PLS regression[J], Analytica Chimica Acta, 2003, 480:23-37
[88] Sekkal M, Huvenne J P, Legrand P, Sombret B, Mollet J, Mouradi-Givernaud A, Verdus M, Direct structural identification of polysaccharides from red algae by FT-IR microspectromtery Ⅰ: Localization of agar in Gracilaria verrucosa sections[J], MikrochimActa, 1993,(121): 110
[89] 史升耀，刘万庆，李智恩，用~(13)C-NMR分析几种中国麒麟菜卡拉胶，海洋与湖沼，1987，18(3)，265-272
[90] 裴小娟，苏敏，田东萍，等．硒化合物与EGFR／MAPK信号传导相关性的初步研究[J]．肿瘤防治杂志，2004，11(12)：1233-1238
[91] 刘德新，黄宏度，吴一慧，王尤富，非离子表面活性剂对石油氧化皂及其复配体系界面活性的影响[J]，汉江石油学院学报，1997，19(4)：50-53
[2] 王长云，耿美玉，管华诗。海洋药物研究进展与发展趋势[J]，中国新药杂志，2005，14(3)，278-282
[3] 盛建春，杨方美，胡秋辉，海藻多糖生物活性研究[J]，食品科学，2005，26(3)，262-264
[4] 张翼，李晓明，王斌贵，海藻生物活性物质研究的回顾与展望[J]，世界科技研究与发展，2005，27(05)：56-63
[5] 盛建春，杨方美，胡秋辉，海藻多糖生物活性研究[J]，食品科学，2005，26(3)，262-264
[6] 王长云，管华诗，多糖抗病毒作用研究进展Ⅰ．多糖抗病毒作用[J]，生物工程进展，2000，20(1)：17-20
[7] Woo E R, Kim W S, Kim Y S, Virus-cell fusion inhibitory activity for the polysaccharides from various Korean edible clams[J], Arch. Pharm. Res., 2001, 24(6): 514—517
[8] Huheihel M, Ishanu V, Tal J, et al, Activity of Porphyridium sp. Polysaccharide against herpes simplex viruses in vitro and in vivo[J], Biochem. Biophys. Methods, 2002, 50:189-200
[9] 辛现良，耿美玉，李桂玲，管华诗，李泽琳，海洋硫酸多糖911体外对HIV-1的作用研究[J]，中国海洋药物，2000，(3)：8-10
[10] Fumihiko T, Rieko N, Tomoko K, et al, Lentinan aufrnent skin reation induced by bradykinin: its correlation with vascular dilatation and hemorrhage responses and antitumor activities[J], Internat immunopharmacol, 1995,17(6):465-474
[11] Kim K H, Lee Y S, Jung I S, et al, Acidic polysaccharide from Panax ginseng, ginsan, induces Thl cell and macrophage cytokines and generates LAK cells in synergy with rIL-2[J], Planta. Med., 1998, 64(2): 110-115
[12] Koyanagi S, Tanigawa N, Nakagawa H, et al, Oversulfation of fucoidan enhances its anti-angiogenic and antitumor activities[J], Biochemi Pharmacol, 2003, 65:173-179
[13] Katsuhide M, Shin Y, Yuji K, et al, Activity of microbial surface polysaccharides in inhibition of cancer cell adhesion[J], Kagaku Kogaku, 1996, 60(11): 932
[14] Sogawa K, Sumida T, Hamakawa H, et al, Inhibitory effect of a marine microalgal polysaccharide on thetelomerase activity in K562 cells[J], Research Communications in Molecular Pathology and Pharmacology, 1998, 99(3): 259-265
[15] Zhang M, Zhang L N, Cheung Keung P C, et al, Molecular weight and anti-tumor activity of the water-soluble polysaccharides isolated by hot water and ultrasonic treatment from the sclerotia and mycelia of Pleurotus tuber-regium[J], Carbohydrate Polymers, 2004, 56: 123-128.
[16] 师然新，徐祖洪，李恩智，降解的角叉菜多糖的抗肿瘤活性[J]，海洋与湖沼，2000，31(6)：45—48
[17] 季宇彬，张琪，孙红，杨卫东，夏月，羊栖菜多糖对P_(388)小鼠红细胞免疫促进作用的机制研究[J]，中国海洋药物，1998，(2)：14—18
[18] 何雅军，吴谦，文蛤提取物对小鼠DTH反应的双向免疫调节作用[J]，中国海洋药物，1995，(3)：20—22
[19] 杨晓林，孙菊云，许汉年，刘晓慧，张翼伸，张绍伦，褐藻糖胶的免疫调节作用[J]，中国海洋药物，1995，(3)：9—12
[20] 刘茜，焦庆才，刘志礼，螺旋藻多糖及其药理作用的研究进展[J]，中国海洋药物，1998，(1)：48—50
[21] Hu J F, Geng M Y, Zhang J T, et al. An in vitro study of the structure-activity relationships of sulfated polysaccharide from brown algae to its antioxidant effect[J]. J. Asian Nat. Prod. Res., 2001, 3(4): 353—358
[22] Yoon S J, Yu M A, Pyun Y R, et al, The nontoxic mushroom auricularia auricular contains a polysaccharide with anticoagulant activity mediated by antithrombin[J], Thrombosis Research, 2003,(112): 151-158
[23] 刘成梅，付桂明，涂宗财，万茵，百合多糖降血糖功能研究[J]，食品科学，2002，23(6)：113-114
[24] Dawes C J, Stanley N F, Moon R E, Physiological and biochemical studies on the L-carrageenan producing red algae Eucheuma uncinatum Setchell and Gardner from the gulf of California[J], Bot. Mar., 1977, 20(7): 437-442
[25] 张其标，耳突麒麟菜卡拉胶提取工艺条件的优化选择[J]，食品科学，1999，(4)：34-36
[26] 韩国华，李海霞，吴杨桦，菲律宾刺生麒麟菜提取：ι-卡拉胶的工艺探讨[J]，食品与机械，2001，(3)：31-32
[27] 汤毅珊，赵谋明，高孔荣，麒麟菜卡拉胶提取新工艺的探讨[J]，食品工业科技，1997，(2)：30-34
[28] 林宇野，杨虹，酶法提取多糖的研究[J]，食品与发酵工业，1995，(1)：13-17
[29] 康琰琰，王一飞，朱良，门晓缓，裙带菜茎中硫酸多糖的分离纯化及分析鉴定[J]，中药材，2005，28(9)：769-771
[30] 王兆梅，李琳，郭祀远，蔡妙颜，活性多糖构效关系研究评述[J]，现代化工，2002，22(8)：18-21
[32] 吴立根，毛文君，衍生化多糖的生物活性研究进展[J]，海洋科学，2002，26(5)：23-25
[33] Dace R, Comparison of the anticoagulant action of sulfated and phosphorylated olysaccharides[J], Thrombosts Research,1997, 87(1): 113-121
[34] 邓成华，杨祥良，王雁，顾小曼，硫酸酯化及羧甲基化虎奶多糖的抗病毒作用[J]，中国地方病学杂志，2000，19(4)：251-253
[35] 孟春，郭养浩，石贤爱，陈剑锋，李锋，海藻多糖生物活性及分子修饰[J]，中国生物工程杂志，2004，24(3)：35-39
[36] 刘占峰，孙汉文，多糖的化学修饰研究进展[J]，河北大学学报(自然科学版)2005，25(1)：104-108
[37] 赖萍，林跃鑫，天然多糖分子修饰研究进展[J]，生命的化学，2003，23(3)：183-187
[38] Uryu T, Ikushima N,Katsuraya K, etal. Sulfated alkyl oligoaacharides with potent inhibitory effects on HIV infection.Biochemical Pharmacol, 1992, 43:2385-2392
[39] Tapiero H, Townsend D M, Tew K D, The antioxidant role of selenium and seleno-compounds[J], Biomedicine & Pharmacotherapy, 2003,(57): 134-144
[40] Gladshev V N, Khangulov S V, Stadtman T C, Properties of the selenium and molybdenum-containing nicotinic acid hydroxylase from clostridium barkeri[J], Biochemistry, 1996,(35): 1212-1233
[41] 刘建林，何新乡，陈宏，硒酸酯多糖的特性及其在食品工业中的应用[J]，中国食品添加剂，2003，(5)：80-82
[42] 许峰，张瞳，周波，贾乐，食用蕈菌硒多糖研究进展[J]，生物技术，2006，16(1)：86-88
[43] 戴伟娟，仲伟法，索金良，林志彬，4-硒硫酸酯多糖对人肝癌细胞Bel／7402DNA RNA及蛋白质合成的影响[J]，济宁医学院学报，2001，24(1)：18-19
[44] 陈展寰，等．无机化学新论选读[M]，西安：陕西人民出版社，1998
[45] 张新宇，王番，绿色巴夫藻硒多糖的提取、分离与纯化[J]，海洋与湖沼，2000，31(6)：643-645
[46] Yoshida M, Fukunaga K, Tsuchita H, et al. Anevaluation of the bio-availability of selenium in high-selenium yeast[J], J-Nutr-Sci-Vitaminol-Tokyo, 1999, 45(1): 119-128
[47] Spolar M R, Schaffer E M, Beelman R B, et al, Selenium-enriched Agaricus bisporus mushrooms suppress 7,12-dimethlybenz[a] anthracene bioactivation in mammary tissue[J], Cancer Lett., 1999, 138(122): 145～150
[48] 尚德静，王孝敏，许永廷，田星，安悦，SeGLP-1的分离纯化及硒活性结构分析[J]，食用菌学报，2002，9(1)：22-27
[49] 郑文杰，贺鸿志，黄峙，杨芳，螺旋藻富集和转化硒研究进展[J]，中国生物工程杂志，2003，23(1)：57-60
[50] 唐家骏，芮海风，硒化卡拉胶的制法，专利CN 1014522B
[51] 龚晓钟，欧阳政，硒化黄芪多糖制备条件及其结构的研究[J]，天然产物研究与开发，1997，10(2)：26-32
[52] 龚晓钟，欧阳政，硒化葡聚糖结构测定及药效研究[J]，深圳大学学报(理工版)，1998，15(1)：70-74
[53] 李光友，缪锦来，刘发义，硒多糖化合物及其制备方法，专利CN 1121414C
[54] 崔乔，尚德静，邹霞，硒多糖的研究进展[J]，中国生化药物杂志，2003，24(3)，155-157
[55] 尚德静，崔乔，高林，灵芝硒多糖SeGLP-1抑制小鼠肝腹水癌作用的研究[J]，食用菌学报，2001，3：34-38
[56] Combs G F, Garbisu C, Lee B C, et al, Bioavailability of selenium accumulated by selenite-reducing bacteria[J], Biological Trace Element Research, 1996, 52(3), 209-225
[57] Nuttall K L, Elemental selenium and glutathione reductase[J], Medical hypotheses, 1985, 16(2): 155-158
[58] 高学云，张劲松，张立德，朱茂祥，纳米红色元素硒对小鼠的免疫功能的调节作用[J]，中国公共卫生，2000，16(5)：421-422
[59] 张劲松，高学云，张立德，朱茂祥，杨陟华，蛋白质分散的纳米红色元素硒的延缓衰老作用[J]，营养学报，2000，22(3)：219-222
[60] Gao X Y, Zhang J S, Zhang L D, Hollow sphere selenium nanoparticles: their in-vitro anti hydroxyl radical effect[J], Advanced Materials, 2002, 14(4): 290-293
[61] Huang B, Zhang J S, Hou J W, Chen C, Free radical scavenging efficiency of Nano-Se in vitro[J], Free Radical Biology & Medicine, 2003, 35(7): 805-813
[62] Zhang JinSong, Gao XueYun, Zhang LiDe, Bao YongPing, Biological effects of a nano red elemental selenium[J], BioFactors, 2001, 15(1):27-38
[63] Zhang J S, Wang H L, Bao Y P, Zhang L D, Nano red elemental selenium has no size effect in the induction of seleno-enzymes in both cultured cells and mice[J], Life Sciences, 2004, 75(2): 237-244
[64] Johnson J A, Saboungi M L, Thiyagarajan P, Csencsits R, Meisel D, Selenium Nanoparticles: A Small-Angle Neutron Scattering Study[J], Journal of Physical Chemistry B, 1999, 103(1): 59-63
[65] Abdelouas A, Gong W L, Lutze W, et al, Using cytochrome C3 to make selenium nanowires[J], Chem. Mater., 2000,12:1510～1512
[66] Gates B, Yin Y D, Xia Y N, et al, A solution-phase approach to the synthesis of uniform nanowires of crystalline selenium with lateral dimensional in the range of 10-30nm[J], J. Am. Chem. Soc., 2000, 122:12582～12583
[67] Mees D R, Pysto W, Tarcha P J, Formation of selenium Colloids Using Sodium Ascorbate as the Reducing Agent[J], Journal of Colloid and Interface Science, 1995, 170:254-260
[68] Gao X Y, Zhang J S, Zhang L D, Hollow Sphere Selenium Nanoparticles: There In-vitro Anti Hydroxyl Radical Effect[J], Adv. Mater., 2002, 14(4): 290-293
[69] 张胜义，张娟，刘明珠，朱俊杰，陈洪渊，PVP存在下液液界面生长法制备硒纳米线[J]，无机化学学报，2005，21(1)：137-141
[70] Gates B, Mayers B, Grossman A, et al, A Sonochemical Approach to the Synthesis of Crystalline Selenium Nanowires in Solutions and on Solid Supports[J],Adv. Mater., 2000, 14(23): 1749-1752
[71] Gates B, Mayers B, Cattle B, et al, Synthesis and Characterization of Uniform Nanowires of Trigonal Selenium Whose Diameters Could Be Controlled in the Range of 10-800nm[J]. Adv. Funct. Mater., 2002, 12(3): 219-227
[72] Bunker C E, Harruff B A, Pathak P, et al, Formation of cadmium sulfide nanoparticles in reverse micelles: Extreme sensitivity to preparation procedure[J], Langmuir,2004,20(13): 5642-5644
[73] Palla B J, Shah D O, Stabilization of high ionic strength slurries using surfactant mixtures: molecular factors that determine optimal stability[J], Journal of Colloid and Interface Science, 2002, 256:143-152
[74] Tian B Z, Liu X Y, Zhang Z D, Tu B, Zhao D Y, Syntheses of High-Quality Mesoporous Materials Directed by Blends of Nonionic Amphiphiles under Nonaqueous Conditions[J], Journal of Solid State Chemistry, 2002, 167(2): 324-329
[75] Wang Y D, Ma C L, Sun X D, Li H D, Synthesis and characterization of ordered hexagonal and cubic mesoporous tin oxides via mixed-surfactant templates route[J], Journal of Colloid and Interface Science, 2005, 296:627-631
[76] Shen Q, Wei H, Wang L C, et al, Crystallization and aggregation behaviors of calcium carbonate in the presence of poly(vinylpyrrolidone) and sodium dodecyl sulfate[J], Journal of Physical Chemistry B, 2005, 109(39): 18342-18347
[77] Yu J G, Zhao X F, Cheng B, et al, Controlled synthesis of calcium carbonate in a mixed aqueous solution of PSMA and CTAB[J], Journal of Solid State Chemistry, 2005, 178(3): 861-867
[78] Kang S H, Hirasawa I, Kim W S, et al, Morphological control of calcium carbonate crystallized in reverse micelle system with anionic surfactants SDS and AOT[J], Journal of Colloid and Interface Science, 2005, 288(2): 496-502
[79] Somasundaran P, Changes in micelle compositions and monomer concentrations in mixed surfactant solutions[J], Langmuir, 1996, 12:5790-5795
[80] Md Emdadul Haque, Akhil Ranjandas, Satya priya moulik J, Mixed micelles of sodium deoxycholate and polyoxyethylene sorbitan monooleate(Tween80)[J], Journal of Colloid and Interface Science, 1999, 217:1-7
[81] 高景镰，麒麟菜的开发利用，中国野生植物[J]，1991，(2)：31-33
[82] 吴风山，吴涤，光度法测定黑木耳中微量硒[J]，光谱实验室，2004，21(4)：719-721
[83] 刘庆堂，李锐增，李坚，紫外分光光度法测定龙胆中微量元素硒[J]，广东药学，2001，11(6)30-32
[84] 高静娴，谷学新，赵晓倩，微孔滤膜富集分光光度法测定痕量硒[J]，首都师范大学学报，1996，17(1)：60-64
[85] 杨金辉，杨江柳，胡鄂明，邓喆，柳建祥，电感耦合等离子体发射光谱法(ICP-AES)测定铀矿石中微量钪、硒、铼[J]，南华大学学报(理工版)，2004，18(4)：83-85
[86] 梁智渊，岑颖洲，叶绍明，伍秋明，琼枝和异枝麒麟菜中硫酸酯基多糖不同提取方法的化学分析比较[J]，暨南大学学报，2005，26(3)：380-385
[87] Prado-Fernandez J, Rodriguez-Vazquez J A, Tojo E, Andrade J M, Quantitation of κ-, ι-and λ-carrageenans by mid-infrared spectroscopy and PLS regression[J], Analytica Chimica Acta, 2003, 480:23-37
[88] Sekkal M, Huvenne J P, Legrand P, Sombret B, Mollet J, Mouradi-Givernaud A, Verdus M, Direct structural identification of polysaccharides from red algae by FT-IR microspectromtery Ⅰ: Localization of agar in Gracilaria verrucosa sections[J], MikrochimActa, 1993,(121): 110
[89] 史升耀，刘万庆，李智恩，用~(13)C-NMR分析几种中国麒麟菜卡拉胶，海洋与湖沼，1987，18(3)，265-272
[90] 裴小娟，苏敏，田东萍，等．硒化合物与EGFR／MAPK信号传导相关性的初步研究[J]．肿瘤防治杂志，2004，11(12)：1233-1238
[91] 刘德新，黄宏度，吴一慧，王尤富，非离子表面活性剂对石油氧化皂及其复配体系界面活性的影响[J]，汉江石油学院学报，1997，19(4)：50-53