黑盖木层孔菌菌丝体多糖分离纯化、结构解析及活性研究
|
摘要
黑盖木层孔菌(Phellinus nigricans)菌丝体经水提得菌丝体多糖PNW,通过正交试验确定菌丝体多糖的最优提取工艺为提取温度100℃,浸提4次,每次2h,浸提比(质量比)1:30,各因素影响大小的次序是:提取次数>浸提比>浸提时间。
GC分析PNW的组成:由甘露糖和葡萄糖构成(摩尔比2.68:1.00)。
经冻融分级、醇沉分级、脱蛋白、DEAE-纤维素柱层析和Sepharose CL-6B柱层析等方法,从PNW中纯化出均一多糖PNW1。HPLC检验为单一峰,平均分子量为33 KDa。GC分析PNW1由半乳糖、甘露糖、葡萄糖和少量的阿拉伯糖和岩藻糖组成,摩尔比为:8.77:6.44:3.26:1.00:1.35。
对PNW1的结构分析采用了部分酸水解、高碘酸氧化、Smith降解、甲基化等化学分析方法及IR、GC、GC/MS、~(13)C-NMR等仪器分析方法。确定了PNW1结构的重复单元。
初步考察了PNW和PNW1对小鼠体外脾淋巴细胞增殖的影响,结果显示PNW和PNW1有增强脾淋巴细胞增殖的作用,PNW1的作用效果优于PNW。PNW和PNW1还可以促进ConA诱导的T淋巴细胞增殖作用和LPS诱导的B淋巴细胞增殖作用。
The mycelia polysaccharides(PNW)were extracted from the mycelia of Phellinus nigricans using boiling water. The extracted condition of PNW was optimized by orthogonal experiment. The optimum condition was that the temperature 100℃, four times, two hours per times and the diffusion ratio 1:30.
GC analysis indicated that the PNW was composed of Man and Glc with the molar ratio of 2.68: 1.00.
PNW1 was purified from PNW using a combination of techniques, such as: ethanol fractional precipitation, deproteinization, DEAE cellulose column chromatography and Sepharose CL-6B column chromatography. HPLC analysis manifested that PNW1 was uniform fraction and the molecular weight was about 33 thousand Dalton. GC analysis indicated that the PNW1 was composed of Gal, Man, Glc, Ara and Xyl with the molar ratio of 8.77:6.44:3.26:1.00:1.35.
The chemical and instrumental analysis methods such as : Partial acid hydrolysis, Periodate oxidation, Smith degradation, IR, GC, GC/MS and ~(13)C-NMR were used on the structure analysis of PNW1.Then the repeated structural unit of PNW1 was determined.
Initially investigated the effects on mouse leukomonocyte proliferation in vitro by PNW and PNW1. The results were that both PNW and PNW1 had significant effects on leukomonocyte proliferation and can promote the effects on the leukomonocyte proliferation stimulated by Con A or LPS. The effects of PNW1 on leukomonocyte proliferation were better than PNW.
GC分析PNW的组成:由甘露糖和葡萄糖构成(摩尔比2.68:1.00)。
经冻融分级、醇沉分级、脱蛋白、DEAE-纤维素柱层析和Sepharose CL-6B柱层析等方法,从PNW中纯化出均一多糖PNW1。HPLC检验为单一峰,平均分子量为33 KDa。GC分析PNW1由半乳糖、甘露糖、葡萄糖和少量的阿拉伯糖和岩藻糖组成,摩尔比为:8.77:6.44:3.26:1.00:1.35。
对PNW1的结构分析采用了部分酸水解、高碘酸氧化、Smith降解、甲基化等化学分析方法及IR、GC、GC/MS、~(13)C-NMR等仪器分析方法。确定了PNW1结构的重复单元。
初步考察了PNW和PNW1对小鼠体外脾淋巴细胞增殖的影响,结果显示PNW和PNW1有增强脾淋巴细胞增殖的作用,PNW1的作用效果优于PNW。PNW和PNW1还可以促进ConA诱导的T淋巴细胞增殖作用和LPS诱导的B淋巴细胞增殖作用。
The mycelia polysaccharides(PNW)were extracted from the mycelia of Phellinus nigricans using boiling water. The extracted condition of PNW was optimized by orthogonal experiment. The optimum condition was that the temperature 100℃, four times, two hours per times and the diffusion ratio 1:30.
GC analysis indicated that the PNW was composed of Man and Glc with the molar ratio of 2.68: 1.00.
PNW1 was purified from PNW using a combination of techniques, such as: ethanol fractional precipitation, deproteinization, DEAE cellulose column chromatography and Sepharose CL-6B column chromatography. HPLC analysis manifested that PNW1 was uniform fraction and the molecular weight was about 33 thousand Dalton. GC analysis indicated that the PNW1 was composed of Gal, Man, Glc, Ara and Xyl with the molar ratio of 8.77:6.44:3.26:1.00:1.35.
The chemical and instrumental analysis methods such as : Partial acid hydrolysis, Periodate oxidation, Smith degradation, IR, GC, GC/MS and ~(13)C-NMR were used on the structure analysis of PNW1.Then the repeated structural unit of PNW1 was determined.
Initially investigated the effects on mouse leukomonocyte proliferation in vitro by PNW and PNW1. The results were that both PNW and PNW1 had significant effects on leukomonocyte proliferation and can promote the effects on the leukomonocyte proliferation stimulated by Con A or LPS. The effects of PNW1 on leukomonocyte proliferation were better than PNW.
引文
[1] 陈惠黎主编,王克夷副主编.糖复合物的结构和功能[M] .第 1 版.上海医科大学出版社,1997.
[2] 张树政主编.糖生物学与糖生物工程[M].第 1 版.清华大学出版社,2002.
[3] Borchers A T,Stern J S,Hackman R M,et al.Mushrooms,tumors and immunity [J].Proc Soc Exp Biol Med ,1999,221 (4):281-293.
[4] Ooi VE , Liu F . Immunomodulation and anti-cancer activity of polysaccharide–protein complexes[J].Curr Med Chem ,2000 ,7 (7):715-729.
[5] 王晓炜,程光宇,吴京燕,等.超微粉碎和普通粉碎对柳松菇多糖的提取及凝胶柱层析分离的研究[J] .南京师大学报(自然科学版),2006,29 (1):66-70.
[6] 李艳辉,王琦.真菌多糖的生物活性与构效关系研究评介[J] .吉林农业大学学报,2002,24 (2) :70-74.
[7] 王小东,毛慧玲,王顺启.真菌多糖的研究进展[J] .江西科学,2005,23 (3):243-246.
[8] 陈红霞.真菌多糖的活性研究进展[J] .生物技术通讯,2005,16 (4) :460-462.
[9] 郭嘉铭.药用真菌的研究与开发概述[J] .中园食用菌,1994,13 (3) :8-10.
[10] 李小定,荣建华,吴谋成.真菌多糖生物活性研究进展[J] .食用菌学报,2002,9 (4):50-58.
[11] 秦俊哲,陈明,陈合,等.食药用真菌多糖的研究现状与展望[J] .中国食用菌,2004,23 (2):6-9.
[12] 孔小卫,江力.蜜环菌胞外多糖对小鼠免疫功能的影响[J].安徽大学学报(自然科学版),2007,31 (1):87-90.
[13] Chihara G,Maeda YY,Hamuro J,et al.Inhibition of mouse sarcoma 180 by polysaccharides from Lentinus edodes[M] .Nature,1969,222:687-688.
[14]杨久根.香菇液体菌种在栽培中的应用[J].中国食用菌,1992,11 (2):25-26.
[15] 贾薇.姬松茸菌丝体多糖提取方法初探[J] .食用菌学报,2003,10 (3):41-44.
[16] 徐任生.天然产物化学[M].第 1 版.北京:科学出版社,1993.469-475.
[17] 史琦云.食用菌多糖提取工艺研究[J] .食品工业科技,2004,25 (2) :98-99.
[18] 林宇野,杨虹.酶法提取银耳多糖的研究[J] .食品与发酵工业,1995 (1):13-17.
[19] Cardoso SM,Silva AMS, Coimbra MA. Structural characterisation of the olive pomacepectic polysaccharide arabinan side chains[J]. Carbohydr. Res, 2002,337,917-924.
[20] 曾凯宏,明建,曾凯芳.真菌多糖的结构与功能[J] .食品科技,2001,6 (4) :65-66.
[21] 胡亚男,王义明,罗国安.质谱在糖分析中的应用[J].药学学报,2000,35 (5):397- 400.
[22] 缪振春,冯锐,周永新,等.四川九节龙中新四糖三萜皂苷结构和核磁共振研究[J] .有机化学,2000,20(3):361-366.
[23] 张雪洪,胡洪波.膜生物反应器连续发酵生产胞外灵芝多糖[J] .高校化学工程学报,2002,16 (6):671- 675.
[24] CU I Y,KIMA DS, PARK KC.Antioxidant effect of Inonotus obliquus [J] .J Ethnopharm acol,2005,96 (1-2):79-85.
[25] ZHANG GL,WANG YH,N IW,et al.Hepato protective role of Ganoderm a lucidum polysaccharide against BCG-induced immune liver injury in mice [J] .World J Gastroen terol,2002,8 (4):728-733.
[26] CHEN YY,CHANG HM.Antiproliferative and differentiating effects of polysaccharide fraction from fu-ling ( Poria cocos) on human leukemic U937 and HL260 cells[J] .Food Chem Toxicol,2004,42 (5) :759-769.
[27] 昊锦文.食用菌的医疗保健作用及其发展趋势[J] .生物学通报,1999,34 (9):18-19.
[28] 陈春霞.羧甲基茯苓多糖促诱生细胞因子的研究[J] .中国食用菌,1998 (2):18-19.
[29] 刘正南.中国药用真菌的现状及种质资源[J] .中国食用菌,1996,15 (5) :20-22.
[30] 吴新君.食(药)用真菌多糖的研究[J] .化学与生物工程,2004,21 (1):14-16.
[31] 刘立,田刚,迟国兴.混合真菌多糖制对小鼠耐力及免疫功能的影响[J] .中国生化药物杂志,1999,20 (4):191-193.
[32] 李小定,荣建华,吴谋成.真菌多糖生物活性研究进展[J] .食用菌学报,2002,4 (9):50-58.
[33] Hikino H, Konno C,Mirin Y,et al.Isolation and hypoglyce micactivity of GanoderansA and B. Glycans of Ganodermalucidum fruitbodies[J] .PlantaMedica,1985,51 (4):339-340.
[34] 杨森.猴头菌提取物对大鼠胃粘膜损伤保护作用的研究[J] .食用菌学报,1999,6 (1):14-17.
[35] 黄伟光,戴文英.灵芝的药效[J] .中国食用菌,1993,12 (4):12-13.
[36] 冯慧琴,杨庆尧,杨晓彤,等.灰树花子实体多糖和菌丝体多糖的比较分析[J] .华东师范大学学报( 自然科学版) ,2001 (3):91-96.
[37] 杜巍,李元瑞,袁静.食药用菌多糖生物活性与结构关系[J] .食用菌,2001 (2):3-5.
[38] 王兆梅,李琳,郭祀远,等.活性多糖构效关系研究评述[J] .现代化工,2002,22 (8):18-22.
[39] Mulloy B,Mourao PAS,Gray E.Structure/function studies of anticoagulant sulphated polysaccharides using NMR[J] .Journal of Biotechnology,2000,77 (1):123-135.
[40] 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.
[41] 张丽萍,张翼伸,孙非,等.硫酸化对金针侧耳多糖构效及生物活性的影响[J] .生物化学与生物物理学报,1994,26 (4):417-421.
[42] Dai YC.Phellinus sensu lato (Aphyllophorales , Hymenochaetaceae) in East Asia [J] .Acta Bot Fennici ,1999 (166):111-115.
[43] Baea J S,Jang KH,Yim H,et al.Polysaccharides isolated from Phellinus gilvus inhibit melanoma growth inmice[J] .Cancer Letters,2005,218 (1):43-52.
[44] 樊锦艳,王秋颖,薛梅,等.桑黄胞外多糖生产培养基的初步研究[J] .2004 (2):93-95.
[45] Andrea T,Borchers,Carl L,et al. The Society for Experimental Biology and Medicine[M].2004.393-406.
[46] 杨菁,黄大斌.碳源和氮源对桑黄菌丝( Phellinus liteus) 生长的影响[J].中国食用菌,2005,24 (4):31-32.
[47] 彭烨城,郑维发,陈才法,等.Phellinus chrysoloma (Fr.) Donk 抗肿瘤作用初步研究[J].中国科技信息,2006 (5):101-102.
[48] 张爽,李红,杨世杰.缝裂层孔菌对小鼠的抑瘤及免疫调节作用[J].吉林大学学报( 医学版),2006,32 (6):964-967.
[49] 郑立军,沈业寿,季俊虬,等.桑黄胞外多糖药理活性的初步研究[J].食品科学,2007,28 (1):318-321.
[50] 温克,陈劲,李红,等.桑黄等四种抗癌药物抗癌活性比较[J].吉林大学学报(医学版),2002,28 (3):247-249.
[51] 吴卫卫,张佩,李淑华,等.桑黄多糖对 S180 的直接抑制作用[J].辽宁医学院学报,2007 ,28 (2):18-20.
[52] 郭军鹏,李红,曲极冰,等.缝裂层孔菌对荷瘤小鼠放、化疗的减毒增效作用[J].吉林大学学报 (医学版),2007,33 (3):518-521.
[53] 李宜明,沈业寿,季俊虬,等.桑黄菌质多糖体外抑瘤及抗环磷酰胺致突变的作用[J].中国科学技术大学学报,2006,36 (7):700-703.
[54] 杨全,胡旭光,王琦,等.药用真菌桑黄菌丝体多糖抗肿瘤作用的研究[J].中国中药杂志,2006,31 (20):1713-1715.
[55] 傅海庆,陈绍军,陈汉清,等.桑黄口服液对小鼠免疫功能的影响[J].福建农林大学学报(自然科学版) ,2006,35 (5):38-54.
[56] 刘安军,任寅,王稳航,等.水溶性 phellinus pini 多糖 PS1 的分离提取及免疫活性的研究[J].现代生物医学进展,2006,6 (11):14-17.
[57] 张万国,胡晋红,蔡溱.桑黄诱生γ_干扰素与抗肝纤维化桑黄预防大鼠肝纤维化作用的实验研究[J].中医药学报,2002,30 (6):22-25.
[58] 张万国,胡晋红,蔡溱.桑黄增强人外周血单个核细胞产生γ_干扰素的研究[J].基层中药杂志,2002,16 (3):5-6.
[59] 张万国,胡晋红,黄瑾.桑黄治疗大鼠肝纤维化实验研究[J].中西医结合肝病杂志,2002,12 (6):348-350.
[60]张万国,胡晋红,蔡溱.桑黄调节细胞因子及其在抗肝纤维化中的意义[J].中国新医药,2003,2 (6):19-20.
[61] 张万国,胡晋红,蔡溱.桑黄对实验性肝纤维化大鼠血液动力学的影响[J].解放军药学学报,2002,18 (6):341-343.
[62] 张万国,胡晋红,蔡溱,等.桑黄对四氯化碳致大鼠肝损伤的保护作用[J].中国药房,2003 ,14 (5):267-269.
[63] 张万国,胡晋红,蔡溱,等.桑黄抗大鼠肝纤维化与抗脂质过氧化[J].中成药,2002,24 (4):281-283.
[64] 张万国,胡晋红,蔡溱,等.桑黄抗大鼠肝纤维化作用实验研究[J].中医药学刊,2001,19 (5):518-519.
[65] 王稳航,李玉,刘安军.松木层孔菌多糖的提取及抗氧化性研究[J].食品研究与开发,2006,27 (11):53-56.
[66] 李宜明,沈业寿,季俊虬,等.桑黄菌质多糖的固态发酵及其抗氧化作用[J].合肥工业大学学报(自然科学版),2006,29(12):1580-1584.
[67] 淡黄木层孔菌子实体提取物治疗呼吸系统炎症、糖尿病, 促进酸性菌的生长[J].国外医药.植物药分册,2006 ,21 (5):223.
[68] 池玉杰,潘学仁.7 种木层孔菌属真菌的培养特性[J].菌物系统 2001,20 (3):378-380.
[69] Hata T.Recent advances in medical and veterinary mycology[M] .Baltimore:University Park Press,1997.299.
[70] Sasaki T,Fujii K,Sugura M,et al.Antitumor polysaccharides from some polyporaceae Ganderma applanatum(Pers) Pat and Phellinus linteus (Berk & Curt) Teng[J] .Chem Pharm Bull ,1971 ,19 (4)::821-826.
[71] Ajith TA , Janardhanan KK.Cytotoxic and antitumor activities of a polypore macrofungus Phellinus rimosus (Berk) Pilat[J] .J Ethnopharmacol,2003,84 (2) :157-162.
[72] Kim HM,Han SB,Oh GT,et al.Stimulation of humoral and cell mediated immunity by polysaccharide from mushroom Phellinus linteus[J] . Int J Immunopharmacol,1996,18 (5):295-303.
[73] Kim GY,Choi GS,Lee SH,et al.Acidic polysaccharide isolated from Phellinus linteus enhances through the up – regulation of nitric oxide and tumor necrosis factor - alpha from peritoneal macrophages[J] .J Ethnopharmacol,2004,95 (1):69-76.
[74] CruegerW , Crueger A . Biotechnology : a textbook of industrial microbiology[M].Sunderland,IL,USA:Sinauer Associates Inc,1990.191–193 .
[75] Blumenkrantz N,Asboe-Hansen G.New method for quantitative determination of uronic acid[M].Anal Biochem,Analytical Biochemistry,1973.484-489.
[76] Wei YA , Fang JN . Determination of purity and molecular weight of polysaccharides by high performance gel permeation chromatography[J].Acta Pharmaceutica Sinica (in Chinese),1989,9 (24):532-536.
[77] 张惟杰.糖复合物生化研究技术[M] .浙江:浙江大学出版社,1999.245-252.
[78] Oades JM.Gas–liquid chromatography of alditol acetates and its application to the analysis of sugars in complex hydrolysates[J].Journal of Chromatography,1967 (28):246-252.
[79] Johnes TM,Albersheim P.A gas chromatographic method for the determination of aldose and uronic acid constituents of plant cell wall polysaccharide[J].Plant Physiology,1972 (49):926-936.
[80] Bao Xing-feng,Wang Xue-Song,Dong Qun,et al.Structural features of immonologicallyactivepolysaccharidesfromGanodermaLucidum[J] . Phytochemistry,2002 (59):175-181.
[81] Milton H Fischer,Yu Nanxiong,Gary R Gray,et al.The gel-forming polysaccharide of psyllium husk [J] .Carbohydrate research,2004 (339) :2009-2017.
[82] Needs PW,Selvendran RR,Selvendran RR.Avoiding oxidative degradation during sodium hydroxide/methyl iodide-mediated carbohydrate methylation in dimethyl sulfoxide[J].Carbohydrate Research,1993 (245) :1-10.
[2] 张树政主编.糖生物学与糖生物工程[M].第 1 版.清华大学出版社,2002.
[3] Borchers A T,Stern J S,Hackman R M,et al.Mushrooms,tumors and immunity [J].Proc Soc Exp Biol Med ,1999,221 (4):281-293.
[4] Ooi VE , Liu F . Immunomodulation and anti-cancer activity of polysaccharide–protein complexes[J].Curr Med Chem ,2000 ,7 (7):715-729.
[5] 王晓炜,程光宇,吴京燕,等.超微粉碎和普通粉碎对柳松菇多糖的提取及凝胶柱层析分离的研究[J] .南京师大学报(自然科学版),2006,29 (1):66-70.
[6] 李艳辉,王琦.真菌多糖的生物活性与构效关系研究评介[J] .吉林农业大学学报,2002,24 (2) :70-74.
[7] 王小东,毛慧玲,王顺启.真菌多糖的研究进展[J] .江西科学,2005,23 (3):243-246.
[8] 陈红霞.真菌多糖的活性研究进展[J] .生物技术通讯,2005,16 (4) :460-462.
[9] 郭嘉铭.药用真菌的研究与开发概述[J] .中园食用菌,1994,13 (3) :8-10.
[10] 李小定,荣建华,吴谋成.真菌多糖生物活性研究进展[J] .食用菌学报,2002,9 (4):50-58.
[11] 秦俊哲,陈明,陈合,等.食药用真菌多糖的研究现状与展望[J] .中国食用菌,2004,23 (2):6-9.
[12] 孔小卫,江力.蜜环菌胞外多糖对小鼠免疫功能的影响[J].安徽大学学报(自然科学版),2007,31 (1):87-90.
[13] Chihara G,Maeda YY,Hamuro J,et al.Inhibition of mouse sarcoma 180 by polysaccharides from Lentinus edodes[M] .Nature,1969,222:687-688.
[14]杨久根.香菇液体菌种在栽培中的应用[J].中国食用菌,1992,11 (2):25-26.
[15] 贾薇.姬松茸菌丝体多糖提取方法初探[J] .食用菌学报,2003,10 (3):41-44.
[16] 徐任生.天然产物化学[M].第 1 版.北京:科学出版社,1993.469-475.
[17] 史琦云.食用菌多糖提取工艺研究[J] .食品工业科技,2004,25 (2) :98-99.
[18] 林宇野,杨虹.酶法提取银耳多糖的研究[J] .食品与发酵工业,1995 (1):13-17.
[19] Cardoso SM,Silva AMS, Coimbra MA. Structural characterisation of the olive pomacepectic polysaccharide arabinan side chains[J]. Carbohydr. Res, 2002,337,917-924.
[20] 曾凯宏,明建,曾凯芳.真菌多糖的结构与功能[J] .食品科技,2001,6 (4) :65-66.
[21] 胡亚男,王义明,罗国安.质谱在糖分析中的应用[J].药学学报,2000,35 (5):397- 400.
[22] 缪振春,冯锐,周永新,等.四川九节龙中新四糖三萜皂苷结构和核磁共振研究[J] .有机化学,2000,20(3):361-366.
[23] 张雪洪,胡洪波.膜生物反应器连续发酵生产胞外灵芝多糖[J] .高校化学工程学报,2002,16 (6):671- 675.
[24] CU I Y,KIMA DS, PARK KC.Antioxidant effect of Inonotus obliquus [J] .J Ethnopharm acol,2005,96 (1-2):79-85.
[25] ZHANG GL,WANG YH,N IW,et al.Hepato protective role of Ganoderm a lucidum polysaccharide against BCG-induced immune liver injury in mice [J] .World J Gastroen terol,2002,8 (4):728-733.
[26] CHEN YY,CHANG HM.Antiproliferative and differentiating effects of polysaccharide fraction from fu-ling ( Poria cocos) on human leukemic U937 and HL260 cells[J] .Food Chem Toxicol,2004,42 (5) :759-769.
[27] 昊锦文.食用菌的医疗保健作用及其发展趋势[J] .生物学通报,1999,34 (9):18-19.
[28] 陈春霞.羧甲基茯苓多糖促诱生细胞因子的研究[J] .中国食用菌,1998 (2):18-19.
[29] 刘正南.中国药用真菌的现状及种质资源[J] .中国食用菌,1996,15 (5) :20-22.
[30] 吴新君.食(药)用真菌多糖的研究[J] .化学与生物工程,2004,21 (1):14-16.
[31] 刘立,田刚,迟国兴.混合真菌多糖制对小鼠耐力及免疫功能的影响[J] .中国生化药物杂志,1999,20 (4):191-193.
[32] 李小定,荣建华,吴谋成.真菌多糖生物活性研究进展[J] .食用菌学报,2002,4 (9):50-58.
[33] Hikino H, Konno C,Mirin Y,et al.Isolation and hypoglyce micactivity of GanoderansA and B. Glycans of Ganodermalucidum fruitbodies[J] .PlantaMedica,1985,51 (4):339-340.
[34] 杨森.猴头菌提取物对大鼠胃粘膜损伤保护作用的研究[J] .食用菌学报,1999,6 (1):14-17.
[35] 黄伟光,戴文英.灵芝的药效[J] .中国食用菌,1993,12 (4):12-13.
[36] 冯慧琴,杨庆尧,杨晓彤,等.灰树花子实体多糖和菌丝体多糖的比较分析[J] .华东师范大学学报( 自然科学版) ,2001 (3):91-96.
[37] 杜巍,李元瑞,袁静.食药用菌多糖生物活性与结构关系[J] .食用菌,2001 (2):3-5.
[38] 王兆梅,李琳,郭祀远,等.活性多糖构效关系研究评述[J] .现代化工,2002,22 (8):18-22.
[39] Mulloy B,Mourao PAS,Gray E.Structure/function studies of anticoagulant sulphated polysaccharides using NMR[J] .Journal of Biotechnology,2000,77 (1):123-135.
[40] 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.
[41] 张丽萍,张翼伸,孙非,等.硫酸化对金针侧耳多糖构效及生物活性的影响[J] .生物化学与生物物理学报,1994,26 (4):417-421.
[42] Dai YC.Phellinus sensu lato (Aphyllophorales , Hymenochaetaceae) in East Asia [J] .Acta Bot Fennici ,1999 (166):111-115.
[43] Baea J S,Jang KH,Yim H,et al.Polysaccharides isolated from Phellinus gilvus inhibit melanoma growth inmice[J] .Cancer Letters,2005,218 (1):43-52.
[44] 樊锦艳,王秋颖,薛梅,等.桑黄胞外多糖生产培养基的初步研究[J] .2004 (2):93-95.
[45] Andrea T,Borchers,Carl L,et al. The Society for Experimental Biology and Medicine[M].2004.393-406.
[46] 杨菁,黄大斌.碳源和氮源对桑黄菌丝( Phellinus liteus) 生长的影响[J].中国食用菌,2005,24 (4):31-32.
[47] 彭烨城,郑维发,陈才法,等.Phellinus chrysoloma (Fr.) Donk 抗肿瘤作用初步研究[J].中国科技信息,2006 (5):101-102.
[48] 张爽,李红,杨世杰.缝裂层孔菌对小鼠的抑瘤及免疫调节作用[J].吉林大学学报( 医学版),2006,32 (6):964-967.
[49] 郑立军,沈业寿,季俊虬,等.桑黄胞外多糖药理活性的初步研究[J].食品科学,2007,28 (1):318-321.
[50] 温克,陈劲,李红,等.桑黄等四种抗癌药物抗癌活性比较[J].吉林大学学报(医学版),2002,28 (3):247-249.
[51] 吴卫卫,张佩,李淑华,等.桑黄多糖对 S180 的直接抑制作用[J].辽宁医学院学报,2007 ,28 (2):18-20.
[52] 郭军鹏,李红,曲极冰,等.缝裂层孔菌对荷瘤小鼠放、化疗的减毒增效作用[J].吉林大学学报 (医学版),2007,33 (3):518-521.
[53] 李宜明,沈业寿,季俊虬,等.桑黄菌质多糖体外抑瘤及抗环磷酰胺致突变的作用[J].中国科学技术大学学报,2006,36 (7):700-703.
[54] 杨全,胡旭光,王琦,等.药用真菌桑黄菌丝体多糖抗肿瘤作用的研究[J].中国中药杂志,2006,31 (20):1713-1715.
[55] 傅海庆,陈绍军,陈汉清,等.桑黄口服液对小鼠免疫功能的影响[J].福建农林大学学报(自然科学版) ,2006,35 (5):38-54.
[56] 刘安军,任寅,王稳航,等.水溶性 phellinus pini 多糖 PS1 的分离提取及免疫活性的研究[J].现代生物医学进展,2006,6 (11):14-17.
[57] 张万国,胡晋红,蔡溱.桑黄诱生γ_干扰素与抗肝纤维化桑黄预防大鼠肝纤维化作用的实验研究[J].中医药学报,2002,30 (6):22-25.
[58] 张万国,胡晋红,蔡溱.桑黄增强人外周血单个核细胞产生γ_干扰素的研究[J].基层中药杂志,2002,16 (3):5-6.
[59] 张万国,胡晋红,黄瑾.桑黄治疗大鼠肝纤维化实验研究[J].中西医结合肝病杂志,2002,12 (6):348-350.
[60]张万国,胡晋红,蔡溱.桑黄调节细胞因子及其在抗肝纤维化中的意义[J].中国新医药,2003,2 (6):19-20.
[61] 张万国,胡晋红,蔡溱.桑黄对实验性肝纤维化大鼠血液动力学的影响[J].解放军药学学报,2002,18 (6):341-343.
[62] 张万国,胡晋红,蔡溱,等.桑黄对四氯化碳致大鼠肝损伤的保护作用[J].中国药房,2003 ,14 (5):267-269.
[63] 张万国,胡晋红,蔡溱,等.桑黄抗大鼠肝纤维化与抗脂质过氧化[J].中成药,2002,24 (4):281-283.
[64] 张万国,胡晋红,蔡溱,等.桑黄抗大鼠肝纤维化作用实验研究[J].中医药学刊,2001,19 (5):518-519.
[65] 王稳航,李玉,刘安军.松木层孔菌多糖的提取及抗氧化性研究[J].食品研究与开发,2006,27 (11):53-56.
[66] 李宜明,沈业寿,季俊虬,等.桑黄菌质多糖的固态发酵及其抗氧化作用[J].合肥工业大学学报(自然科学版),2006,29(12):1580-1584.
[67] 淡黄木层孔菌子实体提取物治疗呼吸系统炎症、糖尿病, 促进酸性菌的生长[J].国外医药.植物药分册,2006 ,21 (5):223.
[68] 池玉杰,潘学仁.7 种木层孔菌属真菌的培养特性[J].菌物系统 2001,20 (3):378-380.
[69] Hata T.Recent advances in medical and veterinary mycology[M] .Baltimore:University Park Press,1997.299.
[70] Sasaki T,Fujii K,Sugura M,et al.Antitumor polysaccharides from some polyporaceae Ganderma applanatum(Pers) Pat and Phellinus linteus (Berk & Curt) Teng[J] .Chem Pharm Bull ,1971 ,19 (4)::821-826.
[71] Ajith TA , Janardhanan KK.Cytotoxic and antitumor activities of a polypore macrofungus Phellinus rimosus (Berk) Pilat[J] .J Ethnopharmacol,2003,84 (2) :157-162.
[72] Kim HM,Han SB,Oh GT,et al.Stimulation of humoral and cell mediated immunity by polysaccharide from mushroom Phellinus linteus[J] . Int J Immunopharmacol,1996,18 (5):295-303.
[73] Kim GY,Choi GS,Lee SH,et al.Acidic polysaccharide isolated from Phellinus linteus enhances through the up – regulation of nitric oxide and tumor necrosis factor - alpha from peritoneal macrophages[J] .J Ethnopharmacol,2004,95 (1):69-76.
[74] CruegerW , Crueger A . Biotechnology : a textbook of industrial microbiology[M].Sunderland,IL,USA:Sinauer Associates Inc,1990.191–193 .
[75] Blumenkrantz N,Asboe-Hansen G.New method for quantitative determination of uronic acid[M].Anal Biochem,Analytical Biochemistry,1973.484-489.
[76] Wei YA , Fang JN . Determination of purity and molecular weight of polysaccharides by high performance gel permeation chromatography[J].Acta Pharmaceutica Sinica (in Chinese),1989,9 (24):532-536.
[77] 张惟杰.糖复合物生化研究技术[M] .浙江:浙江大学出版社,1999.245-252.
[78] Oades JM.Gas–liquid chromatography of alditol acetates and its application to the analysis of sugars in complex hydrolysates[J].Journal of Chromatography,1967 (28):246-252.
[79] Johnes TM,Albersheim P.A gas chromatographic method for the determination of aldose and uronic acid constituents of plant cell wall polysaccharide[J].Plant Physiology,1972 (49):926-936.
[80] Bao Xing-feng,Wang Xue-Song,Dong Qun,et al.Structural features of immonologicallyactivepolysaccharidesfromGanodermaLucidum[J] . Phytochemistry,2002 (59):175-181.
[81] Milton H Fischer,Yu Nanxiong,Gary R Gray,et al.The gel-forming polysaccharide of psyllium husk [J] .Carbohydrate research,2004 (339) :2009-2017.
[82] Needs PW,Selvendran RR,Selvendran RR.Avoiding oxidative degradation during sodium hydroxide/methyl iodide-mediated carbohydrate methylation in dimethyl sulfoxide[J].Carbohydrate Research,1993 (245) :1-10.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |