孔石莼多糖及其铁（Ⅲ）配合物抗辐射有效部位筛选
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摘要
孔石莼是一种广泛分布于西太平洋沿海地区的大型绿藻,资源十分丰富。其具有很高的经济和药用价值,富含蛋白质、氨基酸、糖类等多种营养成分,味道鲜美.孔石莼多糖可增强机体免疫力,具有抗擬血、抗肿瘤、降胆固醇、抗氧化等药用保障功能。
实验中对孔石莼多糖进行水提醇沉,并通过去蛋白、高速离心等对粗多糖进行纯化,预处理后通过100ku和30ku超滤膜对孔石莼多糖进行分级纯化,最终将其分为了≧100ku、30~100ku及≦30ku三个不同分子量段的多糖溶液。经小鼠骨髓细胞微核、肝脾脏系数测定、SOD活性测定及MDA含量测定等多项抗辐射实验,对其抗辐射作用效果进行了比较和筛选,最终发现≦30ku分子量段多糖溶液相较于另两种,其对因辐射引起的骨髓微核率的提升有明显抑制作用(P<0.01),对脾脏系数有较明显的升高(P<0.05),且在小鼠辐照后明显提升了其肝脏中SOD的活性并有效降低了其MDA的含量,通过以上实验结果显示,≦30ku分子量段多糖抗辐射作用强。
取≦30ku分子量段多糖与铁(Ⅲ)离子进行螯合,得到孔石莼多糖铁(Ⅲ)配合物。能谱显示其铁元素的含量较高,原子分光光度法测得其铁含量为27.4%,通过红外光谱显示,其在850cm-1和650cm-1处有特征峰,显示其铁核为β-FeOOH。对多糖及多糖铁(Ⅲ)进行抗辐射实验后发现,两者均有较强的抗辐射作用,但效果差别不大。
对多糖和多糖铁(Ⅲ)辐射实验中的小鼠脾脏进行2-DE蛋白组学研究。分析各组差异点后发现,相对于空白组,辐照对照组中2倍下调的蛋白点有13个,2倍上调的蛋白点有2个;相对于辐照对照组,多糖组中2倍下调蛋白点有5个,2倍上调的蛋白点有14个,多糖铁组中有8个2倍下调的蛋白点,12个2倍上调的蛋白点。从中选取五个差异点进行质谱分析(3502,4705, 5402,5803及6801),发现此五种蛋白分别为78kDa葡萄糖调控蛋白,脯氨酰羟化酶beta,亚基钙网蛋白前体,微管蛋白β5及波形蛋白等。经分析发现,这五种蛋白均与机体免疫及损伤相关,通过对多糖及多糖铁与辐照对照组相比较,发现这两者都在大多蛋白表达上有效果,且其抗辐射效果较好两者差异较小。
coast, and its resources are very rich. Ulva pertusa has a high economic and medicinal value, contains protein, amino acids, sugars and other nutrients,who is very delicious.The polysaccharide of Ulva pertusa can enhance immunity, with the proposed anti-blood, anti-tumor, reduce cholesterol, anti-oxidant and other medical support functions.
The polysaccharide of Ulva pertusa was extracted by water,and precipitated with ethanol, then separated by centrifuagation, the crude polysaccharide extract was obtained. After pretreatment, the polysaccharide of Ulva pertusa was futher purified by 100ku and 30ku membrane, and then the polysaccharide of Ulva pertusa was divided into different molecular weight polysaccharide solution(≧100ku、30~100ku and≦ 30ku). The radioprotective ability of different molecular weight polysaccharide solution were studied using micronucleus assay of mice bone marrow cells, and determination of the spleen index, the activity of SOD and the level of malondialdehyde (MDA). Eventually found the polysaccharide solution less than 30ku showed best ability of radioprotection, it reduced the radiation induced frequency of micronuclei(P<0.01), and increasing the spleen index (P<0.05) and the activity of SOD in the liver after irradiation in mice significantly. The radiation-induced decrease of MDA in the liver by pretreatment wtth polysaccharide (≦30ku). The results show that polysaccharide(≦30ku) possess effitive antiradiation activity.
The polysaccharide of Ulva pertusa(≦30ku) and Fe(Ⅲ) chelation,and got Ulva pertusa polysaccharide iron complex. Result showed high iron(27.4%) of it by atomic spectrophotometry. And in the 850cm-1 and 650cm-1 Office had characteristic peaks by infrared spectroscopy, indicating its iron core wasβ- FeOOH. And also, Ulva pertusa polysaccharide iron complex possess effitive antiradiation activity,compared to the polysaccharide of Ulva pertusa, and the difference of them were not significant.
2-DE study of spleen in mice radiation experiments fed polysaccharide and polysaccharide iron (Ⅲ) show that,compared to control group , 2 proteins are 2-fold up-regulated and 13 2-fold down-regulated in irradiation control group, and compared to irradiation control group , there are 14 2-fold up-regulated proteins , 5 2-fold down-regulated in polysaccharide group and 12 2-fold up-regulated proteins,8 2-fold down-regulated in polysaccharide iron group including 78kDa glucose regulated protein, prolyl hydroxylase beta, subunit calreticulin precursor, micro- tubulinβ5 and vimentin which all were associated with immune and injury.
实验中对孔石莼多糖进行水提醇沉,并通过去蛋白、高速离心等对粗多糖进行纯化,预处理后通过100ku和30ku超滤膜对孔石莼多糖进行分级纯化,最终将其分为了≧100ku、30~100ku及≦30ku三个不同分子量段的多糖溶液。经小鼠骨髓细胞微核、肝脾脏系数测定、SOD活性测定及MDA含量测定等多项抗辐射实验,对其抗辐射作用效果进行了比较和筛选,最终发现≦30ku分子量段多糖溶液相较于另两种,其对因辐射引起的骨髓微核率的提升有明显抑制作用(P<0.01),对脾脏系数有较明显的升高(P<0.05),且在小鼠辐照后明显提升了其肝脏中SOD的活性并有效降低了其MDA的含量,通过以上实验结果显示,≦30ku分子量段多糖抗辐射作用强。
取≦30ku分子量段多糖与铁(Ⅲ)离子进行螯合,得到孔石莼多糖铁(Ⅲ)配合物。能谱显示其铁元素的含量较高,原子分光光度法测得其铁含量为27.4%,通过红外光谱显示,其在850cm-1和650cm-1处有特征峰,显示其铁核为β-FeOOH。对多糖及多糖铁(Ⅲ)进行抗辐射实验后发现,两者均有较强的抗辐射作用,但效果差别不大。
对多糖和多糖铁(Ⅲ)辐射实验中的小鼠脾脏进行2-DE蛋白组学研究。分析各组差异点后发现,相对于空白组,辐照对照组中2倍下调的蛋白点有13个,2倍上调的蛋白点有2个;相对于辐照对照组,多糖组中2倍下调蛋白点有5个,2倍上调的蛋白点有14个,多糖铁组中有8个2倍下调的蛋白点,12个2倍上调的蛋白点。从中选取五个差异点进行质谱分析(3502,4705, 5402,5803及6801),发现此五种蛋白分别为78kDa葡萄糖调控蛋白,脯氨酰羟化酶beta,亚基钙网蛋白前体,微管蛋白β5及波形蛋白等。经分析发现,这五种蛋白均与机体免疫及损伤相关,通过对多糖及多糖铁与辐照对照组相比较,发现这两者都在大多蛋白表达上有效果,且其抗辐射效果较好两者差异较小。
coast, and its resources are very rich. Ulva pertusa has a high economic and medicinal value, contains protein, amino acids, sugars and other nutrients,who is very delicious.The polysaccharide of Ulva pertusa can enhance immunity, with the proposed anti-blood, anti-tumor, reduce cholesterol, anti-oxidant and other medical support functions.
The polysaccharide of Ulva pertusa was extracted by water,and precipitated with ethanol, then separated by centrifuagation, the crude polysaccharide extract was obtained. After pretreatment, the polysaccharide of Ulva pertusa was futher purified by 100ku and 30ku membrane, and then the polysaccharide of Ulva pertusa was divided into different molecular weight polysaccharide solution(≧100ku、30~100ku and≦ 30ku). The radioprotective ability of different molecular weight polysaccharide solution were studied using micronucleus assay of mice bone marrow cells, and determination of the spleen index, the activity of SOD and the level of malondialdehyde (MDA). Eventually found the polysaccharide solution less than 30ku showed best ability of radioprotection, it reduced the radiation induced frequency of micronuclei(P<0.01), and increasing the spleen index (P<0.05) and the activity of SOD in the liver after irradiation in mice significantly. The radiation-induced decrease of MDA in the liver by pretreatment wtth polysaccharide (≦30ku). The results show that polysaccharide(≦30ku) possess effitive antiradiation activity.
The polysaccharide of Ulva pertusa(≦30ku) and Fe(Ⅲ) chelation,and got Ulva pertusa polysaccharide iron complex. Result showed high iron(27.4%) of it by atomic spectrophotometry. And in the 850cm-1 and 650cm-1 Office had characteristic peaks by infrared spectroscopy, indicating its iron core wasβ- FeOOH. And also, Ulva pertusa polysaccharide iron complex possess effitive antiradiation activity,compared to the polysaccharide of Ulva pertusa, and the difference of them were not significant.
2-DE study of spleen in mice radiation experiments fed polysaccharide and polysaccharide iron (Ⅲ) show that,compared to control group , 2 proteins are 2-fold up-regulated and 13 2-fold down-regulated in irradiation control group, and compared to irradiation control group , there are 14 2-fold up-regulated proteins , 5 2-fold down-regulated in polysaccharide group and 12 2-fold up-regulated proteins,8 2-fold down-regulated in polysaccharide iron group including 78kDa glucose regulated protein, prolyl hydroxylase beta, subunit calreticulin precursor, micro- tubulinβ5 and vimentin which all were associated with immune and injury.
引文
1杭金欣主编.浙江沿海藻类与原色图谱[M].浙江科技出版社, 1983:10~11
2王艳梅,李智恩,徐祖洪.孔石莼化学组分和药用活性研究进展[J].海洋科学,2000, 24(3):25~28
3臧笑雪.孔石纯多糖的分离和结构研究[D].青岛:中国海洋大学硕士学位论文, 2006
4苏秀榕,李太武,常少杰.孔石莼营养成分的研究[J].中国海洋药物,1997,61(1): 33~35
5陶平,贺凤伟.大连沿海3种大型速生海藻的营养组成分析[J].中国水产科学,2001, 7(4): 60~63
6 Liquori A M,L.Pizzoferrato,N.Roved,et al.Cellulose from the macroalga’Ulva rigida’of Venice lagoon[J].Chim.Oggi.1991, 9: 57~59
7吴志军,熊慧萍,李智恩.孔石莼脂溶性化学成份的分析[J].海洋科学, 2000, 24(5): 3~4
8纪明侯.海藻化学[M].北京:科学出版社, 1997: 658~684
9张起信.浅谈石莼资源的开发与利用[J].海洋科学, 1998, 3: 1~2
10 Lei Xiaoling, Chen Rong. Elementary study on nutritional compositions of the green alga, Ulva lactucain the South China Sea. Journal of Hainan Normal University(Natural Science). 2003,16(2):79~83
11毛文君,管华诗,李八方.几种海洋生物体内硒含量的测定[J].海洋湖沼通报, 1995, (4): 28~32
12王艳梅,李智恩,牛锡珍等.孔石莼多糖降血脂活性的初步研究[J].中国海洋药物, 2003, 91(2): 33~35
13徐娟华,赵孟辉,谢强明等.海藻石莼降血糖作用的实验研究[J].中国中医药科技, 2002, 9(3): 168~169
14 J.B.Hudson,J.H.Kim,M.K.Lee,et al.Journal of Applied Phycology. 1999,110:427~434
15安卫征.石莼多糖的提取、纯化及生物活性初步研究[D].广州:暨南大学硕士学位论文, 2008
16张颖,岑颖洲,黄日明等.南海七种海藻多糖的抗病毒活性初步研究[J].病毒学报, 2006, 22(4): 282~285
17 Liu Zhenyu,Xie Liyan,Wu Zujian,et al.Purification and characterization of an Anti-TMV protein from a Marin Algae Ulva pertus[J].Acta Phytopathologica Sinica.2005, 35(3): 256~261
18 Hiroyuki Noda,Hideomi Amano,Koiki Arashima,et al.Antitum or activity of marine algae[J]. Hydrobiologia. 1990: 577~584
19 Lee Young Suk,Kim Dong Seuk,Ryu Beung Ho,et al.Aititumor and immuno modulating effects of seaweeds toward sarcoma-180 cells[J].Han'gukYongyang Siklyong Hakhoechi. 1992, 21(5): 544~550
20卞俊,储智勇,鲍蕾蕾等.孔石莼多糖对小鼠免疫功能的影响[J].中国生化药物杂志, 2006, 27(5): 276~279
21徐秀丽,范晓,宋福行.中国经济海藻提取物生物活性[J].海洋与湖沼,2004, 35(1): 55~63
22徐年军,范晓,韩丽君等.海藻乙醇提取物抗菌活性的研究[J].海洋与湖沼, 2002, 3: 265~270
23 Jin Q.,Dong S.L.Comparative studies on the allelopathic effects of two different strains of Ulva pertusa on Heterosigma akashiwo and Alexandrium tamarense[J]. Journal of Experimental Marine Biology and Ecology. 2003, 293: 41~55
24南春容,张海智,董双林.孔石莼水溶性抽提液抑制3种海洋赤潮藻的生长[J].环境科学学报, 2004, 24(4): 702~706
25李丹彤,崔铁军,吕欧等.孔石莼(Ulva pertusa)凝集素的分离纯化及性质的研究[J].中国生物化学与分子生物学报, 2000, 16(6): 774~778
26 Rogers DJ,et al.黄方吕摘.海藻凝集素[J].中国海洋药物, 1991, 38(2):54~56
27 Muto, Shigeaki, Niimura, et al. USPatent5089481, 1992
28中国人民解放军海军后勤部卫生部、上海医药工业研究所合编.中国药用海洋生物[M].上海:上海人民出版社, 1977: 5~6
29刘朝阳,孙晓庆.石莼的综合开发与利用前景[J].饲料广角,2006,17:35~37
30康凯,王长云,王斌等.大型海藻孔石莼化学组分和生物活性研究新进展[J].海洋湖沼通报, 2007, 3: 155~158
31 S.Avallove,J.P. Guiraud, et al. Polysaccharide constituents of coffee-Bean Mueilage. Joumal of food science. 2000, 23(8):1308~1310
32 Peters T, STUIKE-PRILLR, et al. Amontecarlo method for conformational analysis of saccharides[J]. CarRes, 1993, 238: 49
33 B.Ray, M.Lahaye. Cell-wall poylsaccharides from the marine green alga UIva”rigida”(Ulvales,chlorophyta). Chemical structure of ulvna. Carbohydrate Research. 1995b, 274, 313~318
34 Borchers A T, Stem J S, Hackman R M. Mushroom,tumors,immunity[J]. Proc Soc Exp Biol Med. 1999, 221(4): 281~293
35魏小龙,茹祥斌.低分子量地黄多糖对P53基因表达的影响[J].中国药理学报, 1997, 18(5): 471~474
36田庚元.天然多糖的研究和应用(上) [J].上海化工, 2000, (10): 29~31
37张卉,刘长江.姬松茸(Agaricusblazei Murill)生理活性物质的研究进展[J].沈阳农业大学学报, 2003, 34(1): 59~62
38张卉,李长彪,刘长江.姬松茸胞外多糖体外抗氧化活性的研究[J].中国食用菌, 2005, 24(3): 48~49
39李雪华,龙盛京.木耳多糖的提取与抗活性氧的研究[J].食品科学, 2001, 22(3): 26~29
40王雁,杨祥良,邓成华等.羧甲基虎奶多糖的制备及抗氧化性研究[J].生物化学与生物物理进展, 2000, 27(4): 411~413
41庞战军,陈瑗,周玫.云芝多糖对巨噬细胞GPx基因表达的影响[J].生物化学与生物物理学报, 1999, 31(3): 284~288
42刘志辉,孟庆勇.多糖抗幅射研究现状[J].中华放射医学与防护杂志. 2003, 23(1):70
43 M Ito,M Baba,A Sato,et al. Inhibitory effect of dextran sulfate and heparin on the replication of human immun-odeficiency virus(HIV)in vitro[J]. Antiviral Research. 1987, 7: 361~367
44 J Neyts,R Snoeck,D Schols,et al. Sulfated polymers inhibit the interaction of human cytomegalovirus with cell surface heparin sulfate[J]. Virology. 1992, 189(1):48~58
45 ME Gonzalez,B Alarcon,L Carrasco. Polysaccharides as antiviral agents:antiviral activity of carrageenan[J]. An-timicrob Agents Chemother. 1987,31(9):1388~1393
46王长云,管华诗.多糖抗病毒作用研究进展[J].生物工程进展, 2000, 20(1): 17~20
47毛平,夏卉莉.怀牛膝多糖抗凝血作用实验研究[J].时珍国医国药, 2000, 11(12):1075~1076
48 Guyard C,Dehecq E, Tissier JP, et al. Involvement of[beta]-glu-cans in the wide-spectrum antimicrobial activity of Williopsis saturnus var.mrakii MUCL 41968 killer toxin[J]. Molecular Medicine. 2002, 8(11): 686~694
49 Chen YJ, Yang YC. A novel polysaccharide of black soybean promotes myelopoiesis and reconstitutes bone marrow after 5-flurouracil and irradiation-induced myelosuppression[J]. Life Sciences. 2005, 77(4): 400~413
50 JPF Angeli,LR Ribiro,MLC Gonzaga,et al. Protective effeets ofβ-glucan extracted from Agaricus brasiliensis against chemically induced DNA damage inhuman lymphocytes[J]. Cell Biology and Toxicology. 2006, 22(4): 285~291
51 Wang Pei-pei, ZhangYu, Wang Kai-ping et al. Effects of angeliea sinensis Polysaceharide-iron complex on hemolytic anemia and bone marrow injury in mice[J]. Journal of Chinese Pharmaceutical Scienees. 2008, 17: 197~202
52李公斌,王振宇.黑木耳多糖与Fe(Ⅱ)螯合物的制备工艺及红外光谱研究[J].食品工业科技, 2006, 5: 136~138
53秦卫东,马利华等.金针菇多糖螯合亚铁的制备[J].徐州工程学院学报(自然科学版), 2009, 24(4): 15~18
54李玉贤,裴晓红.茶叶多糖铁的合成及其铁含量的测定[J].茶叶科学, 2005, 25(1): 71~74
55李明静,杨静等.怀山药多糖铁(Ⅲ)的组成及性能表征[J].中国中药杂志, 2008,33(16): 2002~2005
56 Maston D R, Thompson R P, Hider R C. Absorption of iron ferric hydroxy pyrannone complex[J]. Br J Nutr. 1994, 71: 203
57 Liu T C,Lin S F,Chang C S, et al. Comparison of a combination ferrous fumarate product and a polysaccharide iron complex as oral treatments of iron deficiency anemia:a Taiwanese study[J]. Int J Hematol. 2004, 80(5): 416~420
58 Sas G, Nemesaszky E, Brauer H. On the therapcutic effects of trivalent and divalent iron deficiency anemia[J]. Arznelm-Forsch/Drug Res. 1984, 34(1): 1575~1579
59 Milligan J R, Aguilera J A, Paglinawan R A, et al. DNA strand break yields after post-high LET irradiation incubation with endonuclease-Ⅲand evidence for hydroxyl radical clustering[J]. Int J Radiat Biol. 2001, 77: 155~164
60 Templeton D M, Cherian M G. Toxicological significance of metallothionein[J]. Methods Enzymol. 1991, 205: 11~24
61 Capizzi R.L.,Oster W.. Protection of normal tissues from the cytotoxic effects of chemotherapy and radiation by amifostine:clinical experiences[J]. European Journal of Cancer(PartA). 1995, 31(1): 8~13
62 Krisch R.E., Ayene I.S., Koch C.J., et al. Simulation of the cellular oxygen effect with an in vitro model system using double strand DNA breaks as an endpoint[J]. International Journal of Radiation oncology’Biology’Physics. 1995, 32(971): 171
63凌笑梅,张娅婕.鳖甲粗多糖对受X射线照射的小鼠的防护作用[J],辐射防护, 1998, 18(1): 57~60
64 Liu Xiaomei, Zhang Hongquan. Effect of Spirulina Platensis Polysaccharide on Hematopoietic Recovery and Related Cytokines in Mice with Transplanted Tumor Treated by Chemotherapy[J]. Chinese Journal of Integrated Traditional and Western Medicine. 2002, 8(2): 130~133
65 Hsin Lin, Dou Mong. Restorative effect of Coriolus versicolor polysaccharides against r-irradiation-induced spleen injury in mice[J]. Chin. Med. J. 1996, 2: 36~41
66 Hu Cheng-jiang, Guo Qing-ping. Protective effect of coriolus versicolor polysaccharides on immunological liver injury in mice[J]. Modern Medicine & Health. 2010, 26(1): 1~2
67 Hui MQ, Ting TZ, Quan BZ, et al. Antioxidant activity of different molecular weight sulfated polysaccharides from Ulva pertusa Kjellm (Chlorophyta) [J]. Journal of Applied Phycology. 2005, 17: 527~534
68赵刚,杨保胜.医学遗传学实验[M].武汉:武汉大学出版社, 1996, 66~70
69曹佳,林真,余争平,等.微核试验——原理、方法及其在人群监测和毒性评价中的应用[M].北京:军事医学科学出版社, 2000: 12~15
70 Pang Q S, Guo B J, Ada K. Radioprotective effect of extract fromSpirulina platensisin mouse bone marrow cells studied by using the micronucleus test [J]. Toxicology Letters. 1989, 48: 165~169
71杜连荣.香叶木素取代物调控静脉血栓蛋白质组及靶蛋白的探索[D].哈尔滨:哈尔滨工业大学硕士论文. 2010
72臧虎.急性脊髓损伤脊髓和脑脊液的蛋白质组学研究[D].长春:吉林大学博士论文. 2004
73王一鸣.桃果实发育硬核期蛋白质双向电泳方法的建立及应用[D].乌鲁木齐:新疆农业大学硕士论文. 2007
74王海凤,钟秀会,李清艳,等. IBV感染雏鸡血清中SOD, GSH-Px活性与MDA含量的研究[ J].畜牧与兽医, 2008, 40(12): 11~14
75 Kuna.P. Duration and degree of radioprotection of WR2721 in mice following its interoeritoneal, intermuscular and subcutaneous adininistration[J]. Radiobioal Radiother. 1983, 24(3):357~364
76 Verhey L J, Sedlacek R. Determination of the radioprotective effects of topical Applications of MEA,WR2721 and N-acetylcysteine on murines skin[J]. Radiat Res. 1980, 93:175~183
77 MarshallP R, Rutherford D. Physical investigations on colloidal iron-dextran complexes[J]. J Colloid Interface Sci . 1971, 37(2): 390
78周晓琳. 78kDa葡萄糖调节蛋白的神经元保护作用研究[D].上海:上海交通大学博士论文. 2007
79 Blackwell GJ, Flower RJ, Nijkamp FP, et a1.Phospholipase A2 activity of guinea-pig isolated perfused 1ungs:stimulation, and inhibitjon by anti-inflammatory steroids[J]. Br J Pharmacol.1978, 62: 79~89
80 Flower RJ.E1eventh Gaddum memorial lecture.Lipocortin and the mechanism ofaction of glucocorticoids.Br J Pharmacol. 1988, 94: 987~1015
81 Osborn M, Weber K. Intermediate filaments: cell-type specific markers in the differentiation and pathology[J]. Cell. 1982, 31: 303
82 Rodrnguez-Pinilla SM, Sarrio D, Honrabo E, et al. Vimentin and laminin expression is associated with basal-like phenotype in both sporadic and BRCA1-associated breast carcinomas[J]. J Clin Pathol. 2007, 60 (9):1006
83 Kim KR, Choi HN, Lee HJ, et al. A peroxisome proliferator-activated receptor gamma antagonist induces vimentin cleavage and inhibits invasion in high-grade hepatocellular carcinoma [J]. Oncol Rep. 2007, 18 (4) :825
84 Singh S, Sadacharan S, Su S, et al. Over expression of Viment in: Role in the invasive phenotype in an androgen independent model of prostate cancer[J]. Cancer Res. 2003, 63(9): 2306
85 Perlson E, Michaelevski I, Kowalsman N, et al.Viment in binding to phosphorylated Erk sterically hinders enzymatic dephosphorylation of the kinase[J]. J Mol Biol. 2006, 364 (5): 938
2王艳梅,李智恩,徐祖洪.孔石莼化学组分和药用活性研究进展[J].海洋科学,2000, 24(3):25~28
3臧笑雪.孔石纯多糖的分离和结构研究[D].青岛:中国海洋大学硕士学位论文, 2006
4苏秀榕,李太武,常少杰.孔石莼营养成分的研究[J].中国海洋药物,1997,61(1): 33~35
5陶平,贺凤伟.大连沿海3种大型速生海藻的营养组成分析[J].中国水产科学,2001, 7(4): 60~63
6 Liquori A M,L.Pizzoferrato,N.Roved,et al.Cellulose from the macroalga’Ulva rigida’of Venice lagoon[J].Chim.Oggi.1991, 9: 57~59
7吴志军,熊慧萍,李智恩.孔石莼脂溶性化学成份的分析[J].海洋科学, 2000, 24(5): 3~4
8纪明侯.海藻化学[M].北京:科学出版社, 1997: 658~684
9张起信.浅谈石莼资源的开发与利用[J].海洋科学, 1998, 3: 1~2
10 Lei Xiaoling, Chen Rong. Elementary study on nutritional compositions of the green alga, Ulva lactucain the South China Sea. Journal of Hainan Normal University(Natural Science). 2003,16(2):79~83
11毛文君,管华诗,李八方.几种海洋生物体内硒含量的测定[J].海洋湖沼通报, 1995, (4): 28~32
12王艳梅,李智恩,牛锡珍等.孔石莼多糖降血脂活性的初步研究[J].中国海洋药物, 2003, 91(2): 33~35
13徐娟华,赵孟辉,谢强明等.海藻石莼降血糖作用的实验研究[J].中国中医药科技, 2002, 9(3): 168~169
14 J.B.Hudson,J.H.Kim,M.K.Lee,et al.Journal of Applied Phycology. 1999,110:427~434
15安卫征.石莼多糖的提取、纯化及生物活性初步研究[D].广州:暨南大学硕士学位论文, 2008
16张颖,岑颖洲,黄日明等.南海七种海藻多糖的抗病毒活性初步研究[J].病毒学报, 2006, 22(4): 282~285
17 Liu Zhenyu,Xie Liyan,Wu Zujian,et al.Purification and characterization of an Anti-TMV protein from a Marin Algae Ulva pertus[J].Acta Phytopathologica Sinica.2005, 35(3): 256~261
18 Hiroyuki Noda,Hideomi Amano,Koiki Arashima,et al.Antitum or activity of marine algae[J]. Hydrobiologia. 1990: 577~584
19 Lee Young Suk,Kim Dong Seuk,Ryu Beung Ho,et al.Aititumor and immuno modulating effects of seaweeds toward sarcoma-180 cells[J].Han'gukYongyang Siklyong Hakhoechi. 1992, 21(5): 544~550
20卞俊,储智勇,鲍蕾蕾等.孔石莼多糖对小鼠免疫功能的影响[J].中国生化药物杂志, 2006, 27(5): 276~279
21徐秀丽,范晓,宋福行.中国经济海藻提取物生物活性[J].海洋与湖沼,2004, 35(1): 55~63
22徐年军,范晓,韩丽君等.海藻乙醇提取物抗菌活性的研究[J].海洋与湖沼, 2002, 3: 265~270
23 Jin Q.,Dong S.L.Comparative studies on the allelopathic effects of two different strains of Ulva pertusa on Heterosigma akashiwo and Alexandrium tamarense[J]. Journal of Experimental Marine Biology and Ecology. 2003, 293: 41~55
24南春容,张海智,董双林.孔石莼水溶性抽提液抑制3种海洋赤潮藻的生长[J].环境科学学报, 2004, 24(4): 702~706
25李丹彤,崔铁军,吕欧等.孔石莼(Ulva pertusa)凝集素的分离纯化及性质的研究[J].中国生物化学与分子生物学报, 2000, 16(6): 774~778
26 Rogers DJ,et al.黄方吕摘.海藻凝集素[J].中国海洋药物, 1991, 38(2):54~56
27 Muto, Shigeaki, Niimura, et al. USPatent5089481, 1992
28中国人民解放军海军后勤部卫生部、上海医药工业研究所合编.中国药用海洋生物[M].上海:上海人民出版社, 1977: 5~6
29刘朝阳,孙晓庆.石莼的综合开发与利用前景[J].饲料广角,2006,17:35~37
30康凯,王长云,王斌等.大型海藻孔石莼化学组分和生物活性研究新进展[J].海洋湖沼通报, 2007, 3: 155~158
31 S.Avallove,J.P. Guiraud, et al. Polysaccharide constituents of coffee-Bean Mueilage. Joumal of food science. 2000, 23(8):1308~1310
32 Peters T, STUIKE-PRILLR, et al. Amontecarlo method for conformational analysis of saccharides[J]. CarRes, 1993, 238: 49
33 B.Ray, M.Lahaye. Cell-wall poylsaccharides from the marine green alga UIva”rigida”(Ulvales,chlorophyta). Chemical structure of ulvna. Carbohydrate Research. 1995b, 274, 313~318
34 Borchers A T, Stem J S, Hackman R M. Mushroom,tumors,immunity[J]. Proc Soc Exp Biol Med. 1999, 221(4): 281~293
35魏小龙,茹祥斌.低分子量地黄多糖对P53基因表达的影响[J].中国药理学报, 1997, 18(5): 471~474
36田庚元.天然多糖的研究和应用(上) [J].上海化工, 2000, (10): 29~31
37张卉,刘长江.姬松茸(Agaricusblazei Murill)生理活性物质的研究进展[J].沈阳农业大学学报, 2003, 34(1): 59~62
38张卉,李长彪,刘长江.姬松茸胞外多糖体外抗氧化活性的研究[J].中国食用菌, 2005, 24(3): 48~49
39李雪华,龙盛京.木耳多糖的提取与抗活性氧的研究[J].食品科学, 2001, 22(3): 26~29
40王雁,杨祥良,邓成华等.羧甲基虎奶多糖的制备及抗氧化性研究[J].生物化学与生物物理进展, 2000, 27(4): 411~413
41庞战军,陈瑗,周玫.云芝多糖对巨噬细胞GPx基因表达的影响[J].生物化学与生物物理学报, 1999, 31(3): 284~288
42刘志辉,孟庆勇.多糖抗幅射研究现状[J].中华放射医学与防护杂志. 2003, 23(1):70
43 M Ito,M Baba,A Sato,et al. Inhibitory effect of dextran sulfate and heparin on the replication of human immun-odeficiency virus(HIV)in vitro[J]. Antiviral Research. 1987, 7: 361~367
44 J Neyts,R Snoeck,D Schols,et al. Sulfated polymers inhibit the interaction of human cytomegalovirus with cell surface heparin sulfate[J]. Virology. 1992, 189(1):48~58
45 ME Gonzalez,B Alarcon,L Carrasco. Polysaccharides as antiviral agents:antiviral activity of carrageenan[J]. An-timicrob Agents Chemother. 1987,31(9):1388~1393
46王长云,管华诗.多糖抗病毒作用研究进展[J].生物工程进展, 2000, 20(1): 17~20
47毛平,夏卉莉.怀牛膝多糖抗凝血作用实验研究[J].时珍国医国药, 2000, 11(12):1075~1076
48 Guyard C,Dehecq E, Tissier JP, et al. Involvement of[beta]-glu-cans in the wide-spectrum antimicrobial activity of Williopsis saturnus var.mrakii MUCL 41968 killer toxin[J]. Molecular Medicine. 2002, 8(11): 686~694
49 Chen YJ, Yang YC. A novel polysaccharide of black soybean promotes myelopoiesis and reconstitutes bone marrow after 5-flurouracil and irradiation-induced myelosuppression[J]. Life Sciences. 2005, 77(4): 400~413
50 JPF Angeli,LR Ribiro,MLC Gonzaga,et al. Protective effeets ofβ-glucan extracted from Agaricus brasiliensis against chemically induced DNA damage inhuman lymphocytes[J]. Cell Biology and Toxicology. 2006, 22(4): 285~291
51 Wang Pei-pei, ZhangYu, Wang Kai-ping et al. Effects of angeliea sinensis Polysaceharide-iron complex on hemolytic anemia and bone marrow injury in mice[J]. Journal of Chinese Pharmaceutical Scienees. 2008, 17: 197~202
52李公斌,王振宇.黑木耳多糖与Fe(Ⅱ)螯合物的制备工艺及红外光谱研究[J].食品工业科技, 2006, 5: 136~138
53秦卫东,马利华等.金针菇多糖螯合亚铁的制备[J].徐州工程学院学报(自然科学版), 2009, 24(4): 15~18
54李玉贤,裴晓红.茶叶多糖铁的合成及其铁含量的测定[J].茶叶科学, 2005, 25(1): 71~74
55李明静,杨静等.怀山药多糖铁(Ⅲ)的组成及性能表征[J].中国中药杂志, 2008,33(16): 2002~2005
56 Maston D R, Thompson R P, Hider R C. Absorption of iron ferric hydroxy pyrannone complex[J]. Br J Nutr. 1994, 71: 203
57 Liu T C,Lin S F,Chang C S, et al. Comparison of a combination ferrous fumarate product and a polysaccharide iron complex as oral treatments of iron deficiency anemia:a Taiwanese study[J]. Int J Hematol. 2004, 80(5): 416~420
58 Sas G, Nemesaszky E, Brauer H. On the therapcutic effects of trivalent and divalent iron deficiency anemia[J]. Arznelm-Forsch/Drug Res. 1984, 34(1): 1575~1579
59 Milligan J R, Aguilera J A, Paglinawan R A, et al. DNA strand break yields after post-high LET irradiation incubation with endonuclease-Ⅲand evidence for hydroxyl radical clustering[J]. Int J Radiat Biol. 2001, 77: 155~164
60 Templeton D M, Cherian M G. Toxicological significance of metallothionein[J]. Methods Enzymol. 1991, 205: 11~24
61 Capizzi R.L.,Oster W.. Protection of normal tissues from the cytotoxic effects of chemotherapy and radiation by amifostine:clinical experiences[J]. European Journal of Cancer(PartA). 1995, 31(1): 8~13
62 Krisch R.E., Ayene I.S., Koch C.J., et al. Simulation of the cellular oxygen effect with an in vitro model system using double strand DNA breaks as an endpoint[J]. International Journal of Radiation oncology’Biology’Physics. 1995, 32(971): 171
63凌笑梅,张娅婕.鳖甲粗多糖对受X射线照射的小鼠的防护作用[J],辐射防护, 1998, 18(1): 57~60
64 Liu Xiaomei, Zhang Hongquan. Effect of Spirulina Platensis Polysaccharide on Hematopoietic Recovery and Related Cytokines in Mice with Transplanted Tumor Treated by Chemotherapy[J]. Chinese Journal of Integrated Traditional and Western Medicine. 2002, 8(2): 130~133
65 Hsin Lin, Dou Mong. Restorative effect of Coriolus versicolor polysaccharides against r-irradiation-induced spleen injury in mice[J]. Chin. Med. J. 1996, 2: 36~41
66 Hu Cheng-jiang, Guo Qing-ping. Protective effect of coriolus versicolor polysaccharides on immunological liver injury in mice[J]. Modern Medicine & Health. 2010, 26(1): 1~2
67 Hui MQ, Ting TZ, Quan BZ, et al. Antioxidant activity of different molecular weight sulfated polysaccharides from Ulva pertusa Kjellm (Chlorophyta) [J]. Journal of Applied Phycology. 2005, 17: 527~534
68赵刚,杨保胜.医学遗传学实验[M].武汉:武汉大学出版社, 1996, 66~70
69曹佳,林真,余争平,等.微核试验——原理、方法及其在人群监测和毒性评价中的应用[M].北京:军事医学科学出版社, 2000: 12~15
70 Pang Q S, Guo B J, Ada K. Radioprotective effect of extract fromSpirulina platensisin mouse bone marrow cells studied by using the micronucleus test [J]. Toxicology Letters. 1989, 48: 165~169
71杜连荣.香叶木素取代物调控静脉血栓蛋白质组及靶蛋白的探索[D].哈尔滨:哈尔滨工业大学硕士论文. 2010
72臧虎.急性脊髓损伤脊髓和脑脊液的蛋白质组学研究[D].长春:吉林大学博士论文. 2004
73王一鸣.桃果实发育硬核期蛋白质双向电泳方法的建立及应用[D].乌鲁木齐:新疆农业大学硕士论文. 2007
74王海凤,钟秀会,李清艳,等. IBV感染雏鸡血清中SOD, GSH-Px活性与MDA含量的研究[ J].畜牧与兽医, 2008, 40(12): 11~14
75 Kuna.P. Duration and degree of radioprotection of WR2721 in mice following its interoeritoneal, intermuscular and subcutaneous adininistration[J]. Radiobioal Radiother. 1983, 24(3):357~364
76 Verhey L J, Sedlacek R. Determination of the radioprotective effects of topical Applications of MEA,WR2721 and N-acetylcysteine on murines skin[J]. Radiat Res. 1980, 93:175~183
77 MarshallP R, Rutherford D. Physical investigations on colloidal iron-dextran complexes[J]. J Colloid Interface Sci . 1971, 37(2): 390
78周晓琳. 78kDa葡萄糖调节蛋白的神经元保护作用研究[D].上海:上海交通大学博士论文. 2007
79 Blackwell GJ, Flower RJ, Nijkamp FP, et a1.Phospholipase A2 activity of guinea-pig isolated perfused 1ungs:stimulation, and inhibitjon by anti-inflammatory steroids[J]. Br J Pharmacol.1978, 62: 79~89
80 Flower RJ.E1eventh Gaddum memorial lecture.Lipocortin and the mechanism ofaction of glucocorticoids.Br J Pharmacol. 1988, 94: 987~1015
81 Osborn M, Weber K. Intermediate filaments: cell-type specific markers in the differentiation and pathology[J]. Cell. 1982, 31: 303
82 Rodrnguez-Pinilla SM, Sarrio D, Honrabo E, et al. Vimentin and laminin expression is associated with basal-like phenotype in both sporadic and BRCA1-associated breast carcinomas[J]. J Clin Pathol. 2007, 60 (9):1006
83 Kim KR, Choi HN, Lee HJ, et al. A peroxisome proliferator-activated receptor gamma antagonist induces vimentin cleavage and inhibits invasion in high-grade hepatocellular carcinoma [J]. Oncol Rep. 2007, 18 (4) :825
84 Singh S, Sadacharan S, Su S, et al. Over expression of Viment in: Role in the invasive phenotype in an androgen independent model of prostate cancer[J]. Cancer Res. 2003, 63(9): 2306
85 Perlson E, Michaelevski I, Kowalsman N, et al.Viment in binding to phosphorylated Erk sterically hinders enzymatic dephosphorylation of the kinase[J]. J Mol Biol. 2006, 364 (5): 938