蝙蝠蛾拟青霉多糖的结构及活性研究
|
摘要
本论文采用水煮、醇沉的方法,得到蝙蝠蛾拟青霉粗多糖CPHP,CPHP经冻融分级、酶-Sevag法联合脱蛋白、Sepharose CL-6B柱层析和HPLC制备等方法进行纯化,得到PHP,PHP经HPLC分析,为均一组分多糖,分子量为28 kDa。
GC分析表明,PHP主要由Man、Gal、Glc和Rha四种单糖组成,其摩尔比为13.36:8.18:3.59:1。PHP经部分酸水解反应发现,PHP的主链由Man和Gal两种单糖组成,糖链末端或支链主要由Glc、Gal和少量Rha组成。高碘酸氧化及Smith降解结果表明,1→、1→6键型是PHP的主要连接键型,同时还存在1→、1→2、1→2,6、1→4、1→4,6键型。
PHP甲基化分析结果表明,PHP中共存在6种键型,其中Man检出1→2、1→2,6两种键型;Gal检出1→和1→6两种键型;Glc检出1→6键型;Rha检出1→2键型,除Rha外,检出的键型都能被高碘酸氧化,与高碘酸氧化结果一致。经计算,四种单糖的摩尔比为Man:Gal:Glc:Rha = 15.26:9.10:3.63:1,与气象色谱结果基本一致。
CPHP对淋巴细胞的增殖及转化的影响结果表明,CPHP在100-400μg/ml浓度下,均能促进淋巴细胞的增殖,并且随着浓度的增加,促进作用越明显;在100-200μg/ml时,CPHP可以促进ConA诱导的T淋巴细胞的增殖,而对LPS诱导的B淋巴细胞的增殖无明显影响。
CPHP对S180肉瘤的抑制作用结果:PHP在50、100、200 mg/kg浓度下的抑瘤率分别为0.32±0.15、0.44±0.18、0.34±0.24,而环磷酰胺阳性组的抑瘤率为0.75±0.10,该结果表明,PHP对S180肉瘤具有抑制作用,但抑制作用不如阳性药物环磷酰胺;与此同时,100 mg/kg CPHP给药组小鼠的脾指数显著高于阴性对照组(p<0.05),而CPHP给药组小鼠的胸腺指数显著低于生理盐水组;ELISA法检测血清中TNF-α水平结果表明,与阴性对照相比,CPHP在100 mg/kg和200 mg/kg给药后,荷瘤小鼠血清中的TNF-α水平明显增加,并且随着给药浓度的增加,血清中TNF-α水平越高。说明CPHP可以促进S180荷瘤小鼠TNF-α的生成。
The polysaccharide of CPHP was isolated from Paecilomyces hepialis by hot water.The water-extracted polysaccharide CPHP was fractionated by frozen-thawed, deproteinated by Sevag method, and purified by anion-exchange chromatography on a column of DEAE-Sepharose CL-6B and gel filtration chromatography on a column of Sepharose CL-6B. Then fraction were obtained and named as PHP.HPLC analysis showed that the polysaccharide of PHP was homogeneous components, and the average molecular weights was 28kDa.
GC analysis showed that PHP was composed of Man,Gla,Glc,and small content of Rha. The molar ratio was 13.36:8.18:3.59:1. The partial acid hydrolysis of PHP showed that the main chain of PHP was composed of Man and Gal, The end of sugar chain or branched-chain was composed of Glc,Gal and Rha. Periodate oxidation and Smith degradation analysis suggested that PHP was mainly contained 1→、1→6 bond type.In addition, there were 1→、1→2、1→2,6、1→4、1→4,6 in PHP.
The methylation analysis applied for PHP show that there were six bond type in PHP.Man was detected in 1→2 and 1→6;Gal was detected in 1→and 1→2,6;Glc was detected in 1→6; Rha was detected in 1→2.All of them could be Oxidized by Periodate. The molar ratio of Man,Gal,Glc and Rha was 2.90:1.73:0.69:0.19.It was are in agreement with GC analysis.
CPHP was cultivated with lymphocyte cells.Between 100μg/ml to 400μg/ml, CPHP could induce lymphoctes proliferation,and the higher the concentration, the effect is more obvious. At the dose of 100 and 200μg/ml, CPHP can promote the ConA-induced proliferation of T lymphocytes, However, it did not affect the LPS-induced proliferation of B lymphocytes.
At the dose of 50,100,and 200 mg/kg,CPHP could inhibit tumor growth in S180 tumor-bearing mice .tumor inhibition rates were 0.32±0.15,0.44±0.18,and 0.34±0.24.they are lower than the positive control group of Cyclophosphamide.meanwhile, Spleen index of CPHP treated group was significantly higher than the negative control group at the dose of 100mg/kg.however, Among the dose of 50,100,and 200 mg/kg,thymus index of CPHP treated group was lower than the negative control group.
ELISA analysis of TNF-αshowed that CPHP could increase the expression of tumor necrosis factor-alpha (TNF-α) in serum at the dose of 100mg/kg and 200mg/kg,and the higher the concentration, the effect is more obvious. These results indicated that the tumor inhibitory activity of the CPHP may be related to the stimulation of immune system, which has a great significance in therapeutics of the tumor growth.
GC分析表明,PHP主要由Man、Gal、Glc和Rha四种单糖组成,其摩尔比为13.36:8.18:3.59:1。PHP经部分酸水解反应发现,PHP的主链由Man和Gal两种单糖组成,糖链末端或支链主要由Glc、Gal和少量Rha组成。高碘酸氧化及Smith降解结果表明,1→、1→6键型是PHP的主要连接键型,同时还存在1→、1→2、1→2,6、1→4、1→4,6键型。
PHP甲基化分析结果表明,PHP中共存在6种键型,其中Man检出1→2、1→2,6两种键型;Gal检出1→和1→6两种键型;Glc检出1→6键型;Rha检出1→2键型,除Rha外,检出的键型都能被高碘酸氧化,与高碘酸氧化结果一致。经计算,四种单糖的摩尔比为Man:Gal:Glc:Rha = 15.26:9.10:3.63:1,与气象色谱结果基本一致。
CPHP对淋巴细胞的增殖及转化的影响结果表明,CPHP在100-400μg/ml浓度下,均能促进淋巴细胞的增殖,并且随着浓度的增加,促进作用越明显;在100-200μg/ml时,CPHP可以促进ConA诱导的T淋巴细胞的增殖,而对LPS诱导的B淋巴细胞的增殖无明显影响。
CPHP对S180肉瘤的抑制作用结果:PHP在50、100、200 mg/kg浓度下的抑瘤率分别为0.32±0.15、0.44±0.18、0.34±0.24,而环磷酰胺阳性组的抑瘤率为0.75±0.10,该结果表明,PHP对S180肉瘤具有抑制作用,但抑制作用不如阳性药物环磷酰胺;与此同时,100 mg/kg CPHP给药组小鼠的脾指数显著高于阴性对照组(p<0.05),而CPHP给药组小鼠的胸腺指数显著低于生理盐水组;ELISA法检测血清中TNF-α水平结果表明,与阴性对照相比,CPHP在100 mg/kg和200 mg/kg给药后,荷瘤小鼠血清中的TNF-α水平明显增加,并且随着给药浓度的增加,血清中TNF-α水平越高。说明CPHP可以促进S180荷瘤小鼠TNF-α的生成。
The polysaccharide of CPHP was isolated from Paecilomyces hepialis by hot water.The water-extracted polysaccharide CPHP was fractionated by frozen-thawed, deproteinated by Sevag method, and purified by anion-exchange chromatography on a column of DEAE-Sepharose CL-6B and gel filtration chromatography on a column of Sepharose CL-6B. Then fraction were obtained and named as PHP.HPLC analysis showed that the polysaccharide of PHP was homogeneous components, and the average molecular weights was 28kDa.
GC analysis showed that PHP was composed of Man,Gla,Glc,and small content of Rha. The molar ratio was 13.36:8.18:3.59:1. The partial acid hydrolysis of PHP showed that the main chain of PHP was composed of Man and Gal, The end of sugar chain or branched-chain was composed of Glc,Gal and Rha. Periodate oxidation and Smith degradation analysis suggested that PHP was mainly contained 1→、1→6 bond type.In addition, there were 1→、1→2、1→2,6、1→4、1→4,6 in PHP.
The methylation analysis applied for PHP show that there were six bond type in PHP.Man was detected in 1→2 and 1→6;Gal was detected in 1→and 1→2,6;Glc was detected in 1→6; Rha was detected in 1→2.All of them could be Oxidized by Periodate. The molar ratio of Man,Gal,Glc and Rha was 2.90:1.73:0.69:0.19.It was are in agreement with GC analysis.
CPHP was cultivated with lymphocyte cells.Between 100μg/ml to 400μg/ml, CPHP could induce lymphoctes proliferation,and the higher the concentration, the effect is more obvious. At the dose of 100 and 200μg/ml, CPHP can promote the ConA-induced proliferation of T lymphocytes, However, it did not affect the LPS-induced proliferation of B lymphocytes.
At the dose of 50,100,and 200 mg/kg,CPHP could inhibit tumor growth in S180 tumor-bearing mice .tumor inhibition rates were 0.32±0.15,0.44±0.18,and 0.34±0.24.they are lower than the positive control group of Cyclophosphamide.meanwhile, Spleen index of CPHP treated group was significantly higher than the negative control group at the dose of 100mg/kg.however, Among the dose of 50,100,and 200 mg/kg,thymus index of CPHP treated group was lower than the negative control group.
ELISA analysis of TNF-αshowed that CPHP could increase the expression of tumor necrosis factor-alpha (TNF-α) in serum at the dose of 100mg/kg and 200mg/kg,and the higher the concentration, the effect is more obvious. These results indicated that the tumor inhibitory activity of the CPHP may be related to the stimulation of immune system, which has a great significance in therapeutics of the tumor growth.
引文
[1]张丽萍主编,杨建雄副主编.生物化学简明教程[M].高等教育出版社,2009,6,第四版.
[2]张树政主编.糖生物学与糖生物工程[M].清华大学出版社,2002,11,第一版.
[3]阳佛送,李雪华.多糖结构研究的方法和进展[J].食品安全与检测,2008,03:200-203.
[4]刘长福,刘涛,刘秀梅.高效液相色谱法测定香菇多糖的分子量[J].化学工程师,2005,118(7):28-29.
[5]肖炳坤,骆传环等.气相色谱法测定柴胡多糖的组分[J].生命科学仪器,2005,3(3):34-35.
[6]裴丽娟.碱提松木层孔菌菌丝体多糖的结构及活性研究[硕士毕业论文].长春:东北师范大学生命科学学院,2010.
[7]张维杰.糖复合物生化研究技术(第二版)[M].浙江大学出版社,1999.
[8] Nicola Volpi. Milligram- scale preparation and purification of oligosaccharides of defined length possessing the structure of chondroitin from defructosylated capsular polysaccharide K4[J]. Glycobiology,2003,13(9):635- 640.
[9]余杰,崔仕超.鸡腿菇子实体多糖分离纯化和光谱分析[J].汕头大学学报, 2009,01:45-49.
[10]丁士林.波谱分析法[M].第2版,重庆大学出版社,1994.
[11]严衍禄.明代仪器分析[M].北京:北京农业大学出版社,1995.
[12]李海萍,苗在京. NMR技术在多糖研究中的应用[J].安徽农业科学, 2009,37(30):14589—14593.
[13] RINAUDO H P,Two-dimensional NMR spectroscopic studies of fenugreek (Trigonella foenum-graectum L.)galactomannan without chemical fragmentation[J]. Carbohydrate Polymers,2001,45:69-77.
[14]陈志强,金杨.多糖结构分析方法的研究[J].饲料工业,2008,29(6):59-61.
[15]高小荣,刘培勋.多糖结构分析研究进展[J].天津药学, 2003,12,15(6):67-69.
[16] Gamian A.Ulrich J.Defaye J Structural heterogeneity of thesialic-acid-containing oligosaccharides from the lipopolysaccharide of Hafria alvei strain 2 as detected by FAB-MS studies 1998.
[17] Edge C J,R adem acher T W ,W orm ald M R ,et al.Fast Sequencing of O ligosacchrides: the R eagent-array A nalysis M ethod [J]. ProcN atlA cad Sci,1992,89(4):6 338-6 342.
[18]邓云霞,瞿伟菁,曹群华等.金耳胞外多糖的结构分析[J].中草药.2005,36(4):497-498.
[19]严琴琴,周南,何惠林等.小儿川崎病与血小板的膜糖蛋白及代谢产物的相关性研究[J].陕西医学杂志,2003,32 (4):297-300.
[20]李波,芦菲,魏新军.二维核磁共振对羊栖菜褐藻DJL的结构研究[J].波谱学杂志,2006,23(4):419-428.
[21]吴新君,毛培宏等.食(药)用真菌多糖的研究[J].食用菌, 2004( 4):3-5.
[22]刘长喜,钱嘉林.复合食用真菌多糖对荷瘤小鼠、环磷酰胺抑制小鼠T细胞亚群的影响[J].中国食品卫生杂志, 2000 , (5) : 3～6.
[23]麻兵继,冯雅岚等.硫酸化香菇分级多糖抗肿瘤活性的研究[J].中国药师,2010,13(4):451-452.
[24]Barre2Sinoussi F , Chermann JC , Charmaret S ,et al. Isolation ofa Tlymphotropic retrovirus from a patient at risk for acquired immunodeficiency syndrome[J] . Science , 1983 , 220 :868～870.
[25]高焕,杨淑娟等.甘草多糖和甘草酸对NDV感染鸡胚成纤维细胞能力的影响[J].畜牧与兽医,2010,42(11):19-23.
[26]王君敏,胡元亮等. 8种硫酸化多糖对新城疫病毒感染鸡胚成纤维细胞能力的影响[J].南京农业大学学报,2011,34(1): 118- 122.
[27]郜玉钢,于文影等.鹿角盘多糖抗病毒的研究[J].安徽农业科学, 2010,38(22):11857-11858.
[28]王晓燕,李莉等.黑加仑多糖抗氧化活性研究[J].中外医疗,2009,9:6.
[29]余萍,郑立群.茶树菇多糖抗氧化性能的研究[J].食品研究与开发,2009,30(11):36-39.
[30]吕喜茹,郭亮等.姬松茸粗多糖抗氧化作用[J].食用菌学报2010,17(1):69~ 71.
[31]阎家麒,王久一,赵敏.中华猕猴桃多糖的提取及其对自由基的清除作用[J].中国生化药物杂志, 1995,16(1):12-14.
[32]许爱华,陈华圣,王玲等.银杏外种皮多糖对不同状态小鼠血清SOD和MDA形成的影响[J].中国中药杂志,1998,23(12):746-747.
[33]肖湘,俞丽君,邱玉莹等.油柑多糖的提取与清除氧自由基作用研究[J].中国药学杂志, 1998,33(5):279-281.
[34]许彬,程清洲等.竹节参多糖抗衰老作用研究[J].中国医院药学杂志,2009,29(13):1083-1084.
[35]左绍远.螺旋藻多糖对D-半乳糖所致衰老小鼠的作用[J].中国生化药物杂志,1998;19(1):15-8.
[36]徐露,董志.当归多糖抗衰老作用的实验研究[J].激光杂志,2008,29(4):89-90.
[37]方哨征,严茂祥,陈芝芸等.金针菇多糖抗肿瘤作用的研究[J].浙江中医学院学报,1996, 20(5) :34～35.
[38] Miyakoshi H , Aoki T , Mizukoshi M. Acting mechanisms of Lentinan in human ,Ⅱ. Enhancement of non2 specific cell2mediated cytotoxicity as an interferon inducer[J]. Int . J . Immunopharmacol , 1984 , 6(4) :373～379.
[39]周爱武,葛志东,梁君山等.香菇多糖的免疫调节作用[J].中国药理学通报, 1995 ,11 (2) :157～159.
[40]董浦江,姚榛祥. A549细胞免疫抑制因子及香菇多糖对T细胞增殖的影响[J].重庆医科大学学报,2000 , 25 (2) :121～122.
[41]高梅,谢蜀生,秦风华等.猪苓多糖对小鼠免疫功能的增强作用[J].中国免疫学杂志, 1991,7 (3) :185～188.
[42]刘彦平枸杞多糖对小鼠T淋巴细胞亚群和淋巴细胞转化作用的研究[J]青海医学院学报,2000(4).
[43]阐建全,徐荻.甘薯活性多糖抗突变作用的体外实验研究[J].中国粮油学报,2001,11(6):23-27.
[44] Pang Q S.Guo B J.Ada K Radioprotective effect of extract from Spirulina platensis in mouse bone marrow cells studied by using the micronucleus test 1989.
[45]刘葛,焦庆才,刘忠礼.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).
[46]赵容杰,赵正林,王丹等.姬松茸多糖的抗炎作用[J].延边大学医学学报,2004;27(1):19-22.
[47]孔庆胜,魏洪伟.多糖类的生物功能[J].济宁医学院学报,1998,21(3):80-82.
[48] FRANZ G.Polysaccharides in pharmacy: current applications and future concepts[J], Plant Medical, 1989, 55: 493- 497.
[49]邓槐春.海带多糖的药理用[J].中草药, 1987, 18(2): 15.
[50] Tadashi Kibo, et al. Effect of a polysaccharide (TAP) from the fruiting bodies of Tremella auranitia on glucose metabolism in mouse liver[J]. Biotcehnol. Biochem, 2000, 64(2): 417.
[51] NORIKO S, RYOKO G, NAOKO O.Characterization of two acidic polysaccharides having immunological activites from the root of Panax ginseng[J]. Biol Pharm Bull, 1993, 16(1): 22.
[52] Maurice M. Dioscoreatine. The Hypoglyce mic Activity of Dioscorans A,B,C,D and F;Glycans of Dioscorea japonica Rhizophors[J]. Planta, 1986,53(3):168.
[53]陈福君,卢军,张永煜.桑的药理研究:桑叶降血糖有效组分对糖尿病动物糖代谢的影响[J].沈阳药科大学学报, 1996,13(1):24.
[54]顾学裘,顾茂珍.抗衰老、抗痛中药的研究展望[M].北京:中国医药科技出版社, 1989: 22- 33.
[55]李亚娜,彭志英.羊栖草多糖减肥功能评价[J].广州食品工业科技, 2004,20(2):63-64.
[56]肖建辉,梁宗琦,刘爱英.虫草无性型及其相关真菌多糖的研究开发现状[J].药学学报, 2002,37(7):589–592.
[57]戴如琴,李晓明.蝙蝠蛾拟青霉名称的合格化[J].菌物学报,2008,27(5):641-644.
[58]戴如琴,兰江丽,陈伟华.蝙幅蛾拟青霉新种的研究[J].北京农业大学学报,1989,15(2):221—225.
[59]杨金玲,肖薇.蝙蝠蛾拟青霉与冬虫夏草关系的分子系统学研究[J].药学学报,2008,43(4):421—426.
[60]于洪飞,孟宪志.一株虫草头抱菌深层发酵条件的研究[J].微生物学杂志,2006,26(1): 51一53.
[61]姜文侠,孙武岳.蝙蝠蛾拟青霉菌丝体最适发酵条件研究[J].食用菌学报,1999,6(2):38-40.
[62]吴迎春,吴喜民.不同菌种/菌株发酵虫草菌丝体的化学成分比较[J].时珍国医国药,2010,21(5):1060-1061.
[63]陈安徽.陈宏伟.几种虫草无性型菌株深层发酵菌丝体的有效成分分析[J].徐州工程学院学报,2007,2,22(2):5-9.
[64]郑建仙.功能性食品[M].中国轻工业出版社,1995,19.
[65]袁建国.功能因子冬虫夏草多糖的研究开发[J].中国食品添加剂,1999,(2):18-22.
[66]王蕾,于荣敏,张辉等.人工培养蛹虫草多糖的分离纯化及其结构的初步研究[J].中国生化药物杂志,2003,24(1):23—25.
[67] MIYAZAKI T.NAOKO O,YAMADA H.Studies on funsal polysaccharidesXX,galact0maI1nan of Cordyceps sinensis[J]. Chem Phann Bu,1977,25(12):3324—3328.
[68]龚敏,朱勤,王彤等.冬虫夏草多糖的分子结构与免疫活性[J].生物化学杂志,1990,6(6):486—491.
[69]李有贵,范雷法,钟石等.家蚕蛹虫草水溶性多糖的分子结构研究[J].蚕业科学,2008,34(1):169—17.
[70]席昭雁,赵起华.蝙蝠蛾拟青霉菌丝体抗疲劳功能实验研究[J].中国自然医学杂志2006,6,8(2):143-145.
[71]邹伟平,黄明明.蝙蝠蛾拟青霉抗排斥反应机理的初步研究[J].同济医科大学学报,1993,22(4):282-284.
[72]童秋声,黄明明.蝙蝠蛾拟青霉抗艾氏癌作用机理研究[J].同济医科大学学报,1991,20(2):104-106.
[73]元敏,王娜,梁素忍.蝙蝠蛾拟青霉Cs-4对阿霉素肾病大鼠致纤维化因子表达的影响[J].临床肾脏病杂志,2010,1,10(1):35-37.
[74]向明,汤静,张程亮.人工发酵虫草菌丝对链佐霉素致大鼠1型糖尿病的防治作用[J].中国药师2005,8(5):355-358.
[2]张树政主编.糖生物学与糖生物工程[M].清华大学出版社,2002,11,第一版.
[3]阳佛送,李雪华.多糖结构研究的方法和进展[J].食品安全与检测,2008,03:200-203.
[4]刘长福,刘涛,刘秀梅.高效液相色谱法测定香菇多糖的分子量[J].化学工程师,2005,118(7):28-29.
[5]肖炳坤,骆传环等.气相色谱法测定柴胡多糖的组分[J].生命科学仪器,2005,3(3):34-35.
[6]裴丽娟.碱提松木层孔菌菌丝体多糖的结构及活性研究[硕士毕业论文].长春:东北师范大学生命科学学院,2010.
[7]张维杰.糖复合物生化研究技术(第二版)[M].浙江大学出版社,1999.
[8] Nicola Volpi. Milligram- scale preparation and purification of oligosaccharides of defined length possessing the structure of chondroitin from defructosylated capsular polysaccharide K4[J]. Glycobiology,2003,13(9):635- 640.
[9]余杰,崔仕超.鸡腿菇子实体多糖分离纯化和光谱分析[J].汕头大学学报, 2009,01:45-49.
[10]丁士林.波谱分析法[M].第2版,重庆大学出版社,1994.
[11]严衍禄.明代仪器分析[M].北京:北京农业大学出版社,1995.
[12]李海萍,苗在京. NMR技术在多糖研究中的应用[J].安徽农业科学, 2009,37(30):14589—14593.
[13] RINAUDO H P,Two-dimensional NMR spectroscopic studies of fenugreek (Trigonella foenum-graectum L.)galactomannan without chemical fragmentation[J]. Carbohydrate Polymers,2001,45:69-77.
[14]陈志强,金杨.多糖结构分析方法的研究[J].饲料工业,2008,29(6):59-61.
[15]高小荣,刘培勋.多糖结构分析研究进展[J].天津药学, 2003,12,15(6):67-69.
[16] Gamian A.Ulrich J.Defaye J Structural heterogeneity of thesialic-acid-containing oligosaccharides from the lipopolysaccharide of Hafria alvei strain 2 as detected by FAB-MS studies 1998.
[17] Edge C J,R adem acher T W ,W orm ald M R ,et al.Fast Sequencing of O ligosacchrides: the R eagent-array A nalysis M ethod [J]. ProcN atlA cad Sci,1992,89(4):6 338-6 342.
[18]邓云霞,瞿伟菁,曹群华等.金耳胞外多糖的结构分析[J].中草药.2005,36(4):497-498.
[19]严琴琴,周南,何惠林等.小儿川崎病与血小板的膜糖蛋白及代谢产物的相关性研究[J].陕西医学杂志,2003,32 (4):297-300.
[20]李波,芦菲,魏新军.二维核磁共振对羊栖菜褐藻DJL的结构研究[J].波谱学杂志,2006,23(4):419-428.
[21]吴新君,毛培宏等.食(药)用真菌多糖的研究[J].食用菌, 2004( 4):3-5.
[22]刘长喜,钱嘉林.复合食用真菌多糖对荷瘤小鼠、环磷酰胺抑制小鼠T细胞亚群的影响[J].中国食品卫生杂志, 2000 , (5) : 3～6.
[23]麻兵继,冯雅岚等.硫酸化香菇分级多糖抗肿瘤活性的研究[J].中国药师,2010,13(4):451-452.
[24]Barre2Sinoussi F , Chermann JC , Charmaret S ,et al. Isolation ofa Tlymphotropic retrovirus from a patient at risk for acquired immunodeficiency syndrome[J] . Science , 1983 , 220 :868～870.
[25]高焕,杨淑娟等.甘草多糖和甘草酸对NDV感染鸡胚成纤维细胞能力的影响[J].畜牧与兽医,2010,42(11):19-23.
[26]王君敏,胡元亮等. 8种硫酸化多糖对新城疫病毒感染鸡胚成纤维细胞能力的影响[J].南京农业大学学报,2011,34(1): 118- 122.
[27]郜玉钢,于文影等.鹿角盘多糖抗病毒的研究[J].安徽农业科学, 2010,38(22):11857-11858.
[28]王晓燕,李莉等.黑加仑多糖抗氧化活性研究[J].中外医疗,2009,9:6.
[29]余萍,郑立群.茶树菇多糖抗氧化性能的研究[J].食品研究与开发,2009,30(11):36-39.
[30]吕喜茹,郭亮等.姬松茸粗多糖抗氧化作用[J].食用菌学报2010,17(1):69~ 71.
[31]阎家麒,王久一,赵敏.中华猕猴桃多糖的提取及其对自由基的清除作用[J].中国生化药物杂志, 1995,16(1):12-14.
[32]许爱华,陈华圣,王玲等.银杏外种皮多糖对不同状态小鼠血清SOD和MDA形成的影响[J].中国中药杂志,1998,23(12):746-747.
[33]肖湘,俞丽君,邱玉莹等.油柑多糖的提取与清除氧自由基作用研究[J].中国药学杂志, 1998,33(5):279-281.
[34]许彬,程清洲等.竹节参多糖抗衰老作用研究[J].中国医院药学杂志,2009,29(13):1083-1084.
[35]左绍远.螺旋藻多糖对D-半乳糖所致衰老小鼠的作用[J].中国生化药物杂志,1998;19(1):15-8.
[36]徐露,董志.当归多糖抗衰老作用的实验研究[J].激光杂志,2008,29(4):89-90.
[37]方哨征,严茂祥,陈芝芸等.金针菇多糖抗肿瘤作用的研究[J].浙江中医学院学报,1996, 20(5) :34～35.
[38] Miyakoshi H , Aoki T , Mizukoshi M. Acting mechanisms of Lentinan in human ,Ⅱ. Enhancement of non2 specific cell2mediated cytotoxicity as an interferon inducer[J]. Int . J . Immunopharmacol , 1984 , 6(4) :373～379.
[39]周爱武,葛志东,梁君山等.香菇多糖的免疫调节作用[J].中国药理学通报, 1995 ,11 (2) :157～159.
[40]董浦江,姚榛祥. A549细胞免疫抑制因子及香菇多糖对T细胞增殖的影响[J].重庆医科大学学报,2000 , 25 (2) :121～122.
[41]高梅,谢蜀生,秦风华等.猪苓多糖对小鼠免疫功能的增强作用[J].中国免疫学杂志, 1991,7 (3) :185～188.
[42]刘彦平枸杞多糖对小鼠T淋巴细胞亚群和淋巴细胞转化作用的研究[J]青海医学院学报,2000(4).
[43]阐建全,徐荻.甘薯活性多糖抗突变作用的体外实验研究[J].中国粮油学报,2001,11(6):23-27.
[44] Pang Q S.Guo B J.Ada K Radioprotective effect of extract from Spirulina platensis in mouse bone marrow cells studied by using the micronucleus test 1989.
[45]刘葛,焦庆才,刘忠礼.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).
[46]赵容杰,赵正林,王丹等.姬松茸多糖的抗炎作用[J].延边大学医学学报,2004;27(1):19-22.
[47]孔庆胜,魏洪伟.多糖类的生物功能[J].济宁医学院学报,1998,21(3):80-82.
[48] FRANZ G.Polysaccharides in pharmacy: current applications and future concepts[J], Plant Medical, 1989, 55: 493- 497.
[49]邓槐春.海带多糖的药理用[J].中草药, 1987, 18(2): 15.
[50] Tadashi Kibo, et al. Effect of a polysaccharide (TAP) from the fruiting bodies of Tremella auranitia on glucose metabolism in mouse liver[J]. Biotcehnol. Biochem, 2000, 64(2): 417.
[51] NORIKO S, RYOKO G, NAOKO O.Characterization of two acidic polysaccharides having immunological activites from the root of Panax ginseng[J]. Biol Pharm Bull, 1993, 16(1): 22.
[52] Maurice M. Dioscoreatine. The Hypoglyce mic Activity of Dioscorans A,B,C,D and F;Glycans of Dioscorea japonica Rhizophors[J]. Planta, 1986,53(3):168.
[53]陈福君,卢军,张永煜.桑的药理研究:桑叶降血糖有效组分对糖尿病动物糖代谢的影响[J].沈阳药科大学学报, 1996,13(1):24.
[54]顾学裘,顾茂珍.抗衰老、抗痛中药的研究展望[M].北京:中国医药科技出版社, 1989: 22- 33.
[55]李亚娜,彭志英.羊栖草多糖减肥功能评价[J].广州食品工业科技, 2004,20(2):63-64.
[56]肖建辉,梁宗琦,刘爱英.虫草无性型及其相关真菌多糖的研究开发现状[J].药学学报, 2002,37(7):589–592.
[57]戴如琴,李晓明.蝙蝠蛾拟青霉名称的合格化[J].菌物学报,2008,27(5):641-644.
[58]戴如琴,兰江丽,陈伟华.蝙幅蛾拟青霉新种的研究[J].北京农业大学学报,1989,15(2):221—225.
[59]杨金玲,肖薇.蝙蝠蛾拟青霉与冬虫夏草关系的分子系统学研究[J].药学学报,2008,43(4):421—426.
[60]于洪飞,孟宪志.一株虫草头抱菌深层发酵条件的研究[J].微生物学杂志,2006,26(1): 51一53.
[61]姜文侠,孙武岳.蝙蝠蛾拟青霉菌丝体最适发酵条件研究[J].食用菌学报,1999,6(2):38-40.
[62]吴迎春,吴喜民.不同菌种/菌株发酵虫草菌丝体的化学成分比较[J].时珍国医国药,2010,21(5):1060-1061.
[63]陈安徽.陈宏伟.几种虫草无性型菌株深层发酵菌丝体的有效成分分析[J].徐州工程学院学报,2007,2,22(2):5-9.
[64]郑建仙.功能性食品[M].中国轻工业出版社,1995,19.
[65]袁建国.功能因子冬虫夏草多糖的研究开发[J].中国食品添加剂,1999,(2):18-22.
[66]王蕾,于荣敏,张辉等.人工培养蛹虫草多糖的分离纯化及其结构的初步研究[J].中国生化药物杂志,2003,24(1):23—25.
[67] MIYAZAKI T.NAOKO O,YAMADA H.Studies on funsal polysaccharidesXX,galact0maI1nan of Cordyceps sinensis[J]. Chem Phann Bu,1977,25(12):3324—3328.
[68]龚敏,朱勤,王彤等.冬虫夏草多糖的分子结构与免疫活性[J].生物化学杂志,1990,6(6):486—491.
[69]李有贵,范雷法,钟石等.家蚕蛹虫草水溶性多糖的分子结构研究[J].蚕业科学,2008,34(1):169—17.
[70]席昭雁,赵起华.蝙蝠蛾拟青霉菌丝体抗疲劳功能实验研究[J].中国自然医学杂志2006,6,8(2):143-145.
[71]邹伟平,黄明明.蝙蝠蛾拟青霉抗排斥反应机理的初步研究[J].同济医科大学学报,1993,22(4):282-284.
[72]童秋声,黄明明.蝙蝠蛾拟青霉抗艾氏癌作用机理研究[J].同济医科大学学报,1991,20(2):104-106.
[73]元敏,王娜,梁素忍.蝙蝠蛾拟青霉Cs-4对阿霉素肾病大鼠致纤维化因子表达的影响[J].临床肾脏病杂志,2010,1,10(1):35-37.
[74]向明,汤静,张程亮.人工发酵虫草菌丝对链佐霉素致大鼠1型糖尿病的防治作用[J].中国药师2005,8(5):355-358.