不同诱导物提高猪苓甾体含量的研究
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摘要
猪苓[Polyporus umbellatus (Pers.)Fries],在分类学上属于无隔担子菌亚纲、无褶菌目、多孔菌科(Polyporaceae)、多孔菌属(Polyporus)的药用真菌,它的干燥菌核即为猪苓,属于常用中药。猪苓具有利尿、抗癌、抗菌等药理活性,利水渗湿是其主要药理活性。猪苓甾体含量仅次于猪苓多糖,是猪苓的另一主要化学成分,具有诸多生物活性,且大多数无毒副作用,在医药工业上具有广泛的用途和良好的发展前景。此外,猪苓甾体可以作为猪苓药材质量评价指标。因此,猪苓甾体是一种很有开发价值的药用资源,同时猪苓甾体的成功开发也为猪苓资源的充分利用作出贡献甾醇是甾体生物合成途径中的一个重要环节,甾醇类化合物是甾体合成的中间体,在猪苓发酵液中添加甾醇,促进猪苓细胞内甾体合成相关的酶的产生,从而促进甾体的合成,增加猪苓甾体的产量。
本课题选用β-谷甾醇,豆甾醇和胆固醇三种不同甾醇及其混合物作为诱导物,在植物甾醇最佳添加浓度范围内(0.05‰~0.20‰),选择三个浓度梯度,添加在猪苓液体发酵培养基中,并通过加入β-环糊精和超声增加甾醇诱导物的溶解度;于23℃悬浮振荡暗培养10d后,检测干燥猪苓菌丝体和发酵液中甾体积累量的变化,从而筛选出最佳诱导物,及其最佳添加浓度。
通过实验,获得结果如下:
(1)单一诱导物中,β-谷甾醇和胆固醇对猪苓菌丝体的生长都具有促进作用,当β-谷甾醇的浓度为0.020%时,菌丝体的生长状况最好,菌丝体总数目最多,浓度为0.015%时,甾体的含量最高;胆固醇的最佳浓度为0.020%,菌丝体的干重最多,甾体净增长量也是最高的。豆甾醇培养基无论是在菌丝体干重还是甾体的净增长量上,就都不及对照组。三者相比,胆固醇效果最好,菌丝体干重达到最大的浓度和甾体净增长量最高的浓度是一致的。胆固醇浓度为0.020%时,菌丝体甾体含量最大(53.224mg),是三者中诱导效果最好。
(2)混合诱导物对猪苓甾体含量和猪苓菌丝体干重的影响混合诱导物对猪苓菌丝体的生长具有促进作用,菌丝体的总量的已增加,甾体的量都得到不同程度的提高。在菌丝体生长状况方面,E组(β-谷甾醇+胆固醇)在当甾醇浓度达到0.005%时,菌丝体干重最高。含有胆固醇的组(E组、F组G组)猪苓甾醇的净增长量明显高于不含胆固醇的组(D组);除了浓度为0.020%时,甾体净增长量低于其他三组,E组(β-谷甾醇+胆固醇)在其他两个浓度都高于其他组,且当浓度为0.005%时,甾体净增量达到最高(97.338),因此可以说β-谷甾醇与胆固醇混合,是混合诱导物中诱导效果最佳的。
通过研究表明,混合诱导物对猪苓甾体的诱导效果强于单一诱导物对猪苓甾体诱导,胆固醇的诱导效果强于β-谷甾醇和胆固醇,β-谷甾醇与胆固醇的混合物诱导效果最好,当浓度为0.005%,猪苓甾体的净增长量最大,达到97.338mg。猪苓甾体对研究猪苓药用价值,充分利用猪苓资源,甾体药物的合成都具有很重要的意义,所以提高猪苓甾体的含量是很有必要的。
Polyporus [Polyporus umbellatus (Pers.) Fries] is a medicinal Fungi ,which belongs to holobasidi- omycetidae,Aphyllophorales, Polyporaceae, Polyporus in the taxonomy.it shall be Polyporus dry sclerotia, are commonly used in Chinese medicine.Modern medicine study indicated that Polyporus umbellatus Fries have many pharmacological activities,for example, diuresis, anti-imflammation; anti-tumor, anti-radiation and enhancing immune function and so on,which has a good prosperity in apply and exploit in fulture. Therefore, polyporus steroid is a kind of valuable medicinal resources and beneficial to make full use of polyporus resource.steroids is biosynthetic intermediate in steroid synthetic. It can promote enzyme related to steroid synthetic product in cell of Polyporus.
This topic selectedβ-sitosterol, stigmasterol and cholesterol and their mixtures as inducer, add inducers in the range of (0.05‰~ 0.20‰) to liquid fermented medium, by addβ-CD and ultrasound for improving inducer solubility.23℃suspended oscillating culture.Then testing steroid accumulation of Polyporus mycelium and broth. furthemore, three kinds of mixtures of sterol and its optimal concentration, which also compared the induction of sterol best.
The results as follows:
(1) single inducer
β-sitosterol and cholesterol promote the growth of mycelium of Polyporus umbellatus when theβ-sitosterol concentration is 0.020%, the best growth conditions of mycelium, concentration is 0.015%, the highest content of steroids; when cholesterol concentration is 0.020% dry weight of mycelium is biggest, the net increase amount of steroid is the highest. Stigmasterol can not promote Polyporus steroid accumulation.
In a word,when cholesterol concentration is 0.020%, the largest concentration of steroids mycelium (53.224mg), so it is the best effect among the three.
(2) Mixed inducer
Tests show that the hybrid inducer promote on the growth of mycelium of Polyporus .The amount of steroids were increased to varying degrees. Status in the growth of mycelium,when concentration of inducer as 0.005%, E group (β-sitosterol + cholesterol) dry weight of mycelium is the highest. Containing cholesterol group (E group, F Group G Group) Polyporus sterol was significantly higher than the net increase in non-cholesterol group (D group); addition to the concentration of 0.020%, the net increase of steroids was less than the other three groups,E group(β-sitosterol+cholesterol) in the other two levels are higher than the other group,and a concentration of 0.005%,the highest net increase steroid(97.338),it can be said thatβ–sitosterol and cholesterol mixed is the best inducer.
The resule show that the mixed inducer was better than a single inducer .The induction effect of cholesterol is stronger thanβ-sitosterol and cholesterol,β-sitosterol and cholesterol mixture induction the best, when the concentration of 0.005%, Polyporus steroidal largest net increase, reaching 97.338mg. Polyporus steroidal medicinal value of research, make full use of Polyporus resources, the synthesis of steroidal drugs have very important meaning, so to improve the content of Polyporus steroid is necessary.
本课题选用β-谷甾醇,豆甾醇和胆固醇三种不同甾醇及其混合物作为诱导物,在植物甾醇最佳添加浓度范围内(0.05‰~0.20‰),选择三个浓度梯度,添加在猪苓液体发酵培养基中,并通过加入β-环糊精和超声增加甾醇诱导物的溶解度;于23℃悬浮振荡暗培养10d后,检测干燥猪苓菌丝体和发酵液中甾体积累量的变化,从而筛选出最佳诱导物,及其最佳添加浓度。
通过实验,获得结果如下:
(1)单一诱导物中,β-谷甾醇和胆固醇对猪苓菌丝体的生长都具有促进作用,当β-谷甾醇的浓度为0.020%时,菌丝体的生长状况最好,菌丝体总数目最多,浓度为0.015%时,甾体的含量最高;胆固醇的最佳浓度为0.020%,菌丝体的干重最多,甾体净增长量也是最高的。豆甾醇培养基无论是在菌丝体干重还是甾体的净增长量上,就都不及对照组。三者相比,胆固醇效果最好,菌丝体干重达到最大的浓度和甾体净增长量最高的浓度是一致的。胆固醇浓度为0.020%时,菌丝体甾体含量最大(53.224mg),是三者中诱导效果最好。
(2)混合诱导物对猪苓甾体含量和猪苓菌丝体干重的影响混合诱导物对猪苓菌丝体的生长具有促进作用,菌丝体的总量的已增加,甾体的量都得到不同程度的提高。在菌丝体生长状况方面,E组(β-谷甾醇+胆固醇)在当甾醇浓度达到0.005%时,菌丝体干重最高。含有胆固醇的组(E组、F组G组)猪苓甾醇的净增长量明显高于不含胆固醇的组(D组);除了浓度为0.020%时,甾体净增长量低于其他三组,E组(β-谷甾醇+胆固醇)在其他两个浓度都高于其他组,且当浓度为0.005%时,甾体净增量达到最高(97.338),因此可以说β-谷甾醇与胆固醇混合,是混合诱导物中诱导效果最佳的。
通过研究表明,混合诱导物对猪苓甾体的诱导效果强于单一诱导物对猪苓甾体诱导,胆固醇的诱导效果强于β-谷甾醇和胆固醇,β-谷甾醇与胆固醇的混合物诱导效果最好,当浓度为0.005%,猪苓甾体的净增长量最大,达到97.338mg。猪苓甾体对研究猪苓药用价值,充分利用猪苓资源,甾体药物的合成都具有很重要的意义,所以提高猪苓甾体的含量是很有必要的。
Polyporus [Polyporus umbellatus (Pers.) Fries] is a medicinal Fungi ,which belongs to holobasidi- omycetidae,Aphyllophorales, Polyporaceae, Polyporus in the taxonomy.it shall be Polyporus dry sclerotia, are commonly used in Chinese medicine.Modern medicine study indicated that Polyporus umbellatus Fries have many pharmacological activities,for example, diuresis, anti-imflammation; anti-tumor, anti-radiation and enhancing immune function and so on,which has a good prosperity in apply and exploit in fulture. Therefore, polyporus steroid is a kind of valuable medicinal resources and beneficial to make full use of polyporus resource.steroids is biosynthetic intermediate in steroid synthetic. It can promote enzyme related to steroid synthetic product in cell of Polyporus.
This topic selectedβ-sitosterol, stigmasterol and cholesterol and their mixtures as inducer, add inducers in the range of (0.05‰~ 0.20‰) to liquid fermented medium, by addβ-CD and ultrasound for improving inducer solubility.23℃suspended oscillating culture.Then testing steroid accumulation of Polyporus mycelium and broth. furthemore, three kinds of mixtures of sterol and its optimal concentration, which also compared the induction of sterol best.
The results as follows:
(1) single inducer
β-sitosterol and cholesterol promote the growth of mycelium of Polyporus umbellatus when theβ-sitosterol concentration is 0.020%, the best growth conditions of mycelium, concentration is 0.015%, the highest content of steroids; when cholesterol concentration is 0.020% dry weight of mycelium is biggest, the net increase amount of steroid is the highest. Stigmasterol can not promote Polyporus steroid accumulation.
In a word,when cholesterol concentration is 0.020%, the largest concentration of steroids mycelium (53.224mg), so it is the best effect among the three.
(2) Mixed inducer
Tests show that the hybrid inducer promote on the growth of mycelium of Polyporus .The amount of steroids were increased to varying degrees. Status in the growth of mycelium,when concentration of inducer as 0.005%, E group (β-sitosterol + cholesterol) dry weight of mycelium is the highest. Containing cholesterol group (E group, F Group G Group) Polyporus sterol was significantly higher than the net increase in non-cholesterol group (D group); addition to the concentration of 0.020%, the net increase of steroids was less than the other three groups,E group(β-sitosterol+cholesterol) in the other two levels are higher than the other group,and a concentration of 0.005%,the highest net increase steroid(97.338),it can be said thatβ–sitosterol and cholesterol mixed is the best inducer.
The resule show that the mixed inducer was better than a single inducer .The induction effect of cholesterol is stronger thanβ-sitosterol and cholesterol,β-sitosterol and cholesterol mixture induction the best, when the concentration of 0.005%, Polyporus steroidal largest net increase, reaching 97.338mg. Polyporus steroidal medicinal value of research, make full use of Polyporus resources, the synthesis of steroidal drugs have very important meaning, so to improve the content of Polyporus steroid is necessary.
引文
北京中医研究院中药研究所. 1985,猪苓多糖[J].药学通报, 2(2): 74-75
卞志远,李金枝. 1997,猪苓多糖与8AB方案联合治疗小细胞肺癌[J].中国肿瘤生物治疗杂志, 4(3):233
川村清一. 1954原色日本菌图鉴(第一册).东京:东京风间书房,: 130
陈明, 1998,猪苓汤106例验案统计分析[J].中国医药学报. 13 (2): 29-32
陈霞. 2000.猪苓林下栽培技术初报[J].中国林副特产, 3 (2) : 16
陈知本,熊那,朱巧庆.中国药科大学学报.1991,22(4):211-214
董群,方积年. 2001,多糖在医药领域中的应用[J].中国药学杂志, 36(10): 649-651.
戴如琴等. 1979.猪苓菌丝体的液体培养及其抗肿瘤作用的研究[J].新医药学杂志, 2: 19-22
戴肖东,张欣,韩增华等. 2001.野生猪苓菌丝分离培养试验初报[J].食用菌, 23 (6): 10-11
冯青然,付桂兰,孟彬,章育中等. 1987,猪苓多糖分子量和黏度的测定,中药通报, 12(1): 38-40
国家中医药管理局《中华本草》编委会,1999,中华本草[M],上海科学技术出版社, 1-551
国家药典委员会. 2000.中华人民共和国药典[M].北京:化学工业出版社,附录: 53-54
郭顺星,徐锦堂,肖培根. 1996.猪苓生物学特性的研究进展[J].中国药学杂志, 21(9): 515
郭顺星,徐锦堂,王春兰等. 1993.不同年龄的野生与家种猪苓菌核氨基酸及微量元素分析[J].真菌学报, 18(4): 204-206
郭顺星,徐锦堂. 1992.猪苓菌核的营养来源及与蜜环菌的关系.植物学报, 34(8 ) : 576-580
郭顺星,徐锦堂. 1993.蜜环菌侵染猪苓菌核的细胞学研究.植物学报, 35(1): 44-50
郭顺星,徐锦堂,肖培根. 1996.猪苓生物学特性的研究进展[J].中国药学杂志, 21(9): 515
顾若卿. 1987,复方丹参片中三七总皂甙的含量测定[J].中成药研究, 8: 10-11.
高双新,强治国. 1985.猪苓栽培技术的研究[J].陕西中医学院学报, 8(2): 29-31
管伟举,谷克仁.植物甾醇的研究进展[J].粮食与油脂.2006(3):5-9
谷严芳等. 1991,猪苓汤证治规律的研究:古今中外119例统计分析[J]实用中医内科杂志. 5 (1) :14-17
黄芳,蒙义文. 1999 ,活性多糖的研究进展[J].天然产物研究与开发, 11(5): 90-98.
杭共存,刘茂君. 2001,加减猪荃汤治疗肝硬化腹水31例[J].陕西中医. 22(11): 671
韩军花. 2001.植物甾醇的性质、功能及应用[J].国外医学卫生学分册. 28(5):286
黄庆纾, 1998 ,杀伤肿瘤细胞活性的实验研究[J].中国免疫学杂, 14 (2): 109-111.
韩汝城,张维经等. 1980.猪苓与蜜环菌关系的初步研究.中药材科技[J], (2): 6- 8
胡昭庚. 1999.17种药用真菌栽培[M].北京:中国农业出版社: 83-90
姜嘉,巫善明,徐伟民等. 1994,猪苓多糖注射液合并乙型肝炎疫苗抗鸭HBV的作用[J].中华传染病杂志, 12(2): 102.
江苏新医学院编. 1986.中药大辞典[M].上海:上海科学技术出版社. 2191-2192
吕陈峰,陈毅力.王龙刚.杨胜利,王武.2001.利用胆固醇制备胆甾-4-烯-3酮[J].无锡轻工业大学学报,20(5):485~488
刘瑰琦,郑焕春,于长有等. 2001,野外栽培猪苓的实用技术[J].特种经济动植物, 4(6): 28
李静,周立刚,文成敬.2008.真菌甾体化合物的研究进展[J]. (20):165
兰进等. 1994.应用同位素示踪法研究猪苓第二营养源[J].中国药学杂志, 29(7): 394-395
兰进,徐锦堂,贺秀霞. 2001.药用真菌栽培实用技术[M].北京:中国农业出版社,:210-220
李健. 1992.药用菌栽培与加工[M].北京:中国农业科学技术出版社: 45-54
刘荣忠. 1984.药用菌栽培[M].成都:四川科学技术出版社: 67-79
梁生旺,刘伟. 1997.中药制剂定量分析[M] .北京:中国中医药出版社.
刘小丰,陈昌珍,赵玉娟. 1998,抗乙型肝炎免疫核糖核酸联合猪苓多糖治疗慢性乙型肝炎[J].临床荟萃, 13(12): 550
李晓静,冯霞,阳葵.2004.底物的溶解对甾体微生物羟化反应的影响[J].天津大学学报,37(11)941~944
罗英,李梁. 2002.猪苓生长的土壤条件研究[J ].核农学报, 16(2): 115-118
马敬中,张友德. 2000,甾体化合物的分光光度法测定[J].分析化学, 28(3): 390
桑岚. 2000,猪苓汤治疗糖尿病性肾病35例[J].临床报道河南中医药学刊. 15(3)-34-35
邵春杰,徐景达. 1981,人参皂苷的含量测定[J ].中草药, 12
孙凤春,张朴. 1994,猪苓多糖治疗寻常型银屑病[J].中华皮肤科杂志, 27(3):170
山西古县猪苓种植场. 1978.人工种植猪等新成果[J].中药材科技, (1): 19-23
山西省猪苓科研协作组. 1981.猪苓生态环境及生物学特性的调查研究初报[J].中药材科技,(3) :7 -11
孙晔. 2005.猪苓液体发酵及猪苓多糖的研究[D].华东师范大学, 34-36
田庚元,冯宇澄,林颖. 1995 ,植物多糖的研究进展[J].中国中药杂志, 20 (7): 441.
王弘,晁建平,陈文举,杨玥,陈世忠.2009.猪苓药材的质量评价标准研究.中草药[J].40(6)
韦继愫. 2002.植物甾醇的提取工艺研究[D].杭州.浙江大学.
魏群等.中国药理学报, 1983, 4(1), 52
魏群,聂剑初,宋书元,白玉珍,吴国利等. 1983,猪苓多糖对荷肝癌H22小鼠肝糖代谢和肾上腺皮质功能的作用[J],中国药理学报, 4 (1): 52-54
王启祥. 1991,猪苓汤加味治疗尿血症证68例[J].国医论坛. 6(4)-12-12
王秋颖,徐锦堂. 1993.蜜环菌发酵液在猪苓菌发酵过程中的应用[J].中国药学杂志, 28(8): 466-468
王秋颖,徐锦堂. 1997.假单胞杆菌、灵芝等几种菌的发酵液对猪苓菌生长的影响[J].中国药学杂志, 32(8): 459-462
王秋颖,徐锦堂,郭顺星,贺秀霞. 2000.猪苓与蜜环菌营养关系的初步探讨[J].中国中药杂志, 25(8): 472-473
汪友永. 1992,抗乙肝新药H-猪苓多糖注射液[J].中国医院药学杂志, 12(10):447
王艳,吴玉波,张永恒. 1996,猪苓多糖对顺铂增效作用及毒性的影响[J].中医药研究, (5): 60
吴益仙. 1997,猪苓汤治疗老年性癃闭60例[J].四川中医. 15(4)-26-26
王莹莹寇永奎朱建松. 2007,华中师范大学研究生学报[J]. 14(1):143-147
夏洪燕,郭顺星. 2000.猪苓菌丝细胞活性氧的产生及其种类[J].菌物系统, 19(4): 576
夏洪燕,郭顺星. 2001.蜜环菌激发子诱导猪苓活性氧产生及相关酶变化[J].微生物学通报, 28(3):22
徐锦堂. 1997中国药用真菌[M].第一版.北京:中国医科大学,中国协和医科大学,联合出版社.518-534
徐锦堂,郭顺星. 1992.猪苓与蜜环菌的关系.真菌学报, 11(2): 142-145
徐锦堂. 1997.中国药用真菌学[M].北京医科大学·中国协和医科人学联合出版社
徐锦堂. 1997.中国药用真菌学[M].北京:中国医科大学·中国协和医科大学联合出版
阳葵,李晓静,冯霞,尹强,赵海燕.2001.底物的分散和溶解对甾体微生物酶反应的影响[J].微生物学通报,28(6):68~71
严术常,曹望芳,张英华等. 1988,猪苓多糖治疗慢性病毒性肝炎的临床和实验研究[J].中西医结合杂志, 8(3): 141-143
俞志成. 1997.猪苓菌种的制作与栽培加工技术[J].农村科技开发, (2): 28
中国医学科学院药用植物资源开发研究所. 1991,中国药用植物栽培学[M].北京:农业出版社, 1~85
郑虎占等. 1999.中药现代研究与应用[M],学苑出版社, 4159
中华人民共和国卫生部药典委员会. 2000.中国药典部[S ].北京:化学工业出版社,
张娟聪,潘秋月,孟祥河,孙培龙.2008.β-环糊精改善植物甾醇水溶性的研究.中国粮食学报[J]. 24(4):111~112
朱勤,戴如琴,张惟杰.猪苓发酵滤液多糖组分及摩尔比的测定[J].中国中药杂志, 13(9): 544
张新,王志伟. 2000,娑罗子中总皂甙的比色测定[J].天然产物研究与开发, 1(12): 281
周晓燕,顾顺明,张文玉等. 2001.发酵法生产猪苓菌丝体及猪苓多糖的研究[J].工业微生物, 31(4) :1~4
张言良,陈功夫等. 1978.猪苓人工栽培初报[J].陕西新医药, (2): 65~66
赵英勇崔秀明张文斌周家明程显隆张萍孙文基林瑞超.2009.猪苓的化学成分与药理作用研究进展中药材.32(11).11.1785-1787
Chiu S H, ChungT W, Giridhar R, Wu W T. 2004. Immo-bilization ofβ-cyclodextrin in chitosan beads for separation of cholesterol from egg yolk. Food Research International, 37(3): 217–223.
Choi Y H, Yang C H, Kim H W, Jung S. 2001. Molecular modeling studies of theβ-cyclodextrin in monomer and dimer form as hosts for the complexation of cholesterol. Journal of Inclusion Phenomena and Macrocyclic Chemistry, 39(1-2): 71-76. ChP(中国药典).2005.volⅠ(一部):222
Hamuro J , Maeda Y Y, Arai Y, et al. 1971, The significance of the higher structure of the polysaccharides lentinan and pachy-maran with regard to their antitumor activity[J]. Chem BiolInteract , 3(1): 69-71.
Hiroyuki Asanuina, Masaya Kakazu, Mflsahiko Shihata.1998.Synthesis of molecularly imprinted polymerofβ-cyclodextrin for the efficient reeogonization of cholesterol. Suoramolecular science, 5: 417-421.Hitoshi Ishida, et al . 1999, Biol Pharm Bull, 22(11), 1189
IshixucaT, Yaoieta Y, Kikuchi M. 1997. Sterols from the fruit bodies of Grifola from dosa. Chern.Pharm.Bull, 45: 1756-1760
Joachim Rosecke, Wilfried A. K?nig . 2000.Constituents of various wood-rotting basidiomycetes. Phytoc -hemistry, 54(6), 603~610
Kobayshi M, Hayashi T, Hayashi K. 1983. Isolatio and studies on marine Natura products(II). New polyhydroxytated Sterols from the soft coral lobolytum pauciftorum(Ehrenbery). Chem-Pharm Bull, 31, 18-48.
Koji O, Yosulori Y, Noriko M, Masao K. 1996. Sterol Constituents from the Selerotium of Polyporus umbellate Fries. Naturalm edicines, 50( 2): 170—181
Lu W., Adachi I, Kana K, Yasuta A, Toriizuka K, Ueno M. 1985. Chem. Pharm. Bull, 33, 5083
Manohar B, Basappa C, Rao D N, Divakar S. 1998. Response surface studies on cholesterol reduction in egg yolk usingβ-cy-clodextrin. Zeitschrift fuer Lebensmittel-Untersuchung und-Forschung A, 206 (3) : 189–192.
Robert O, WilliamsⅢ, Vompann M, Mongkol S. 1998. Characterizafion of an inclusion complex of β-cycledex-trin and cholesterol by aeration at elevated temperatures. Biochemical Engineering Journal, 22(3): 197-205.
Seoung wook jun.min-soo kim.jeong-soo kim.2007.Perhapation and characterization of simvastafin/hydroxypropy l-β-cyclodextrin inclusion complex using supercritical anti-solvent(SAS) process
Shinjiro Y, Hajime K, Talmaki M, Xin H X, Hajime U. 2005. Cholesterol recovery from inclusion complex T. ohsawa, M. yukawa, C. takao, M. nrayama, AND H. eando. 1992. Studies on constituents of Fruit Body of Polyporus umbellatus Fries and Their cytotoxic Activity. chem-pharm.Bull. 40 (1): 143-147
Takatomi Ohsawa, et al. 1992, Chem Pharm Bull, 40(1), 143
Tayade.Pralhad.Kale.Rajen 2004 drakumar Study of freeze-dried quercefin cyclodextrin binary systems by DSC,FT-IR,X-ray diffraction and SEM analysis
Weete JD. 1989, Structure and function sterols in fungi . Adv Lipid Res, 23: 1152 167 .
XU Jin-tang(徐锦堂). 1997.Chinese Medicinal Fungus(中国药用真菌).Peking Union Medical College and Beijing Medical Univercity Press(北京医科大学.中国协和医科大学联合出版), 150
Yasunori Inaoka. 1994, Chem Pharm Bull, 42(3), 530
You J S, Hau D M, Chen K T, Huang H F.1994.Combined effects of chuling (Polyporus umbellatus) extract and mitomycin C on experimental liver caheer. Am. J. Chin. Med., 22 (1): 19-28.
卞志远,李金枝. 1997,猪苓多糖与8AB方案联合治疗小细胞肺癌[J].中国肿瘤生物治疗杂志, 4(3):233
川村清一. 1954原色日本菌图鉴(第一册).东京:东京风间书房,: 130
陈明, 1998,猪苓汤106例验案统计分析[J].中国医药学报. 13 (2): 29-32
陈霞. 2000.猪苓林下栽培技术初报[J].中国林副特产, 3 (2) : 16
陈知本,熊那,朱巧庆.中国药科大学学报.1991,22(4):211-214
董群,方积年. 2001,多糖在医药领域中的应用[J].中国药学杂志, 36(10): 649-651.
戴如琴等. 1979.猪苓菌丝体的液体培养及其抗肿瘤作用的研究[J].新医药学杂志, 2: 19-22
戴肖东,张欣,韩增华等. 2001.野生猪苓菌丝分离培养试验初报[J].食用菌, 23 (6): 10-11
冯青然,付桂兰,孟彬,章育中等. 1987,猪苓多糖分子量和黏度的测定,中药通报, 12(1): 38-40
国家中医药管理局《中华本草》编委会,1999,中华本草[M],上海科学技术出版社, 1-551
国家药典委员会. 2000.中华人民共和国药典[M].北京:化学工业出版社,附录: 53-54
郭顺星,徐锦堂,肖培根. 1996.猪苓生物学特性的研究进展[J].中国药学杂志, 21(9): 515
郭顺星,徐锦堂,王春兰等. 1993.不同年龄的野生与家种猪苓菌核氨基酸及微量元素分析[J].真菌学报, 18(4): 204-206
郭顺星,徐锦堂. 1992.猪苓菌核的营养来源及与蜜环菌的关系.植物学报, 34(8 ) : 576-580
郭顺星,徐锦堂. 1993.蜜环菌侵染猪苓菌核的细胞学研究.植物学报, 35(1): 44-50
郭顺星,徐锦堂,肖培根. 1996.猪苓生物学特性的研究进展[J].中国药学杂志, 21(9): 515
顾若卿. 1987,复方丹参片中三七总皂甙的含量测定[J].中成药研究, 8: 10-11.
高双新,强治国. 1985.猪苓栽培技术的研究[J].陕西中医学院学报, 8(2): 29-31
管伟举,谷克仁.植物甾醇的研究进展[J].粮食与油脂.2006(3):5-9
谷严芳等. 1991,猪苓汤证治规律的研究:古今中外119例统计分析[J]实用中医内科杂志. 5 (1) :14-17
黄芳,蒙义文. 1999 ,活性多糖的研究进展[J].天然产物研究与开发, 11(5): 90-98.
杭共存,刘茂君. 2001,加减猪荃汤治疗肝硬化腹水31例[J].陕西中医. 22(11): 671
韩军花. 2001.植物甾醇的性质、功能及应用[J].国外医学卫生学分册. 28(5):286
黄庆纾, 1998 ,杀伤肿瘤细胞活性的实验研究[J].中国免疫学杂, 14 (2): 109-111.
韩汝城,张维经等. 1980.猪苓与蜜环菌关系的初步研究.中药材科技[J], (2): 6- 8
胡昭庚. 1999.17种药用真菌栽培[M].北京:中国农业出版社: 83-90
姜嘉,巫善明,徐伟民等. 1994,猪苓多糖注射液合并乙型肝炎疫苗抗鸭HBV的作用[J].中华传染病杂志, 12(2): 102.
江苏新医学院编. 1986.中药大辞典[M].上海:上海科学技术出版社. 2191-2192
吕陈峰,陈毅力.王龙刚.杨胜利,王武.2001.利用胆固醇制备胆甾-4-烯-3酮[J].无锡轻工业大学学报,20(5):485~488
刘瑰琦,郑焕春,于长有等. 2001,野外栽培猪苓的实用技术[J].特种经济动植物, 4(6): 28
李静,周立刚,文成敬.2008.真菌甾体化合物的研究进展[J]. (20):165
兰进等. 1994.应用同位素示踪法研究猪苓第二营养源[J].中国药学杂志, 29(7): 394-395
兰进,徐锦堂,贺秀霞. 2001.药用真菌栽培实用技术[M].北京:中国农业出版社,:210-220
李健. 1992.药用菌栽培与加工[M].北京:中国农业科学技术出版社: 45-54
刘荣忠. 1984.药用菌栽培[M].成都:四川科学技术出版社: 67-79
梁生旺,刘伟. 1997.中药制剂定量分析[M] .北京:中国中医药出版社.
刘小丰,陈昌珍,赵玉娟. 1998,抗乙型肝炎免疫核糖核酸联合猪苓多糖治疗慢性乙型肝炎[J].临床荟萃, 13(12): 550
李晓静,冯霞,阳葵.2004.底物的溶解对甾体微生物羟化反应的影响[J].天津大学学报,37(11)941~944
罗英,李梁. 2002.猪苓生长的土壤条件研究[J ].核农学报, 16(2): 115-118
马敬中,张友德. 2000,甾体化合物的分光光度法测定[J].分析化学, 28(3): 390
桑岚. 2000,猪苓汤治疗糖尿病性肾病35例[J].临床报道河南中医药学刊. 15(3)-34-35
邵春杰,徐景达. 1981,人参皂苷的含量测定[J ].中草药, 12
孙凤春,张朴. 1994,猪苓多糖治疗寻常型银屑病[J].中华皮肤科杂志, 27(3):170
山西古县猪苓种植场. 1978.人工种植猪等新成果[J].中药材科技, (1): 19-23
山西省猪苓科研协作组. 1981.猪苓生态环境及生物学特性的调查研究初报[J].中药材科技,(3) :7 -11
孙晔. 2005.猪苓液体发酵及猪苓多糖的研究[D].华东师范大学, 34-36
田庚元,冯宇澄,林颖. 1995 ,植物多糖的研究进展[J].中国中药杂志, 20 (7): 441.
王弘,晁建平,陈文举,杨玥,陈世忠.2009.猪苓药材的质量评价标准研究.中草药[J].40(6)
韦继愫. 2002.植物甾醇的提取工艺研究[D].杭州.浙江大学.
魏群等.中国药理学报, 1983, 4(1), 52
魏群,聂剑初,宋书元,白玉珍,吴国利等. 1983,猪苓多糖对荷肝癌H22小鼠肝糖代谢和肾上腺皮质功能的作用[J],中国药理学报, 4 (1): 52-54
王启祥. 1991,猪苓汤加味治疗尿血症证68例[J].国医论坛. 6(4)-12-12
王秋颖,徐锦堂. 1993.蜜环菌发酵液在猪苓菌发酵过程中的应用[J].中国药学杂志, 28(8): 466-468
王秋颖,徐锦堂. 1997.假单胞杆菌、灵芝等几种菌的发酵液对猪苓菌生长的影响[J].中国药学杂志, 32(8): 459-462
王秋颖,徐锦堂,郭顺星,贺秀霞. 2000.猪苓与蜜环菌营养关系的初步探讨[J].中国中药杂志, 25(8): 472-473
汪友永. 1992,抗乙肝新药H-猪苓多糖注射液[J].中国医院药学杂志, 12(10):447
王艳,吴玉波,张永恒. 1996,猪苓多糖对顺铂增效作用及毒性的影响[J].中医药研究, (5): 60
吴益仙. 1997,猪苓汤治疗老年性癃闭60例[J].四川中医. 15(4)-26-26
王莹莹寇永奎朱建松. 2007,华中师范大学研究生学报[J]. 14(1):143-147
夏洪燕,郭顺星. 2000.猪苓菌丝细胞活性氧的产生及其种类[J].菌物系统, 19(4): 576
夏洪燕,郭顺星. 2001.蜜环菌激发子诱导猪苓活性氧产生及相关酶变化[J].微生物学通报, 28(3):22
徐锦堂. 1997中国药用真菌[M].第一版.北京:中国医科大学,中国协和医科大学,联合出版社.518-534
徐锦堂,郭顺星. 1992.猪苓与蜜环菌的关系.真菌学报, 11(2): 142-145
徐锦堂. 1997.中国药用真菌学[M].北京医科大学·中国协和医科人学联合出版社
徐锦堂. 1997.中国药用真菌学[M].北京:中国医科大学·中国协和医科大学联合出版
阳葵,李晓静,冯霞,尹强,赵海燕.2001.底物的分散和溶解对甾体微生物酶反应的影响[J].微生物学通报,28(6):68~71
严术常,曹望芳,张英华等. 1988,猪苓多糖治疗慢性病毒性肝炎的临床和实验研究[J].中西医结合杂志, 8(3): 141-143
俞志成. 1997.猪苓菌种的制作与栽培加工技术[J].农村科技开发, (2): 28
中国医学科学院药用植物资源开发研究所. 1991,中国药用植物栽培学[M].北京:农业出版社, 1~85
郑虎占等. 1999.中药现代研究与应用[M],学苑出版社, 4159
中华人民共和国卫生部药典委员会. 2000.中国药典部[S ].北京:化学工业出版社,
张娟聪,潘秋月,孟祥河,孙培龙.2008.β-环糊精改善植物甾醇水溶性的研究.中国粮食学报[J]. 24(4):111~112
朱勤,戴如琴,张惟杰.猪苓发酵滤液多糖组分及摩尔比的测定[J].中国中药杂志, 13(9): 544
张新,王志伟. 2000,娑罗子中总皂甙的比色测定[J].天然产物研究与开发, 1(12): 281
周晓燕,顾顺明,张文玉等. 2001.发酵法生产猪苓菌丝体及猪苓多糖的研究[J].工业微生物, 31(4) :1~4
张言良,陈功夫等. 1978.猪苓人工栽培初报[J].陕西新医药, (2): 65~66
赵英勇崔秀明张文斌周家明程显隆张萍孙文基林瑞超.2009.猪苓的化学成分与药理作用研究进展中药材.32(11).11.1785-1787
Chiu S H, ChungT W, Giridhar R, Wu W T. 2004. Immo-bilization ofβ-cyclodextrin in chitosan beads for separation of cholesterol from egg yolk. Food Research International, 37(3): 217–223.
Choi Y H, Yang C H, Kim H W, Jung S. 2001. Molecular modeling studies of theβ-cyclodextrin in monomer and dimer form as hosts for the complexation of cholesterol. Journal of Inclusion Phenomena and Macrocyclic Chemistry, 39(1-2): 71-76. ChP(中国药典).2005.volⅠ(一部):222
Hamuro J , Maeda Y Y, Arai Y, et al. 1971, The significance of the higher structure of the polysaccharides lentinan and pachy-maran with regard to their antitumor activity[J]. Chem BiolInteract , 3(1): 69-71.
Hiroyuki Asanuina, Masaya Kakazu, Mflsahiko Shihata.1998.Synthesis of molecularly imprinted polymerofβ-cyclodextrin for the efficient reeogonization of cholesterol. Suoramolecular science, 5: 417-421.Hitoshi Ishida, et al . 1999, Biol Pharm Bull, 22(11), 1189
IshixucaT, Yaoieta Y, Kikuchi M. 1997. Sterols from the fruit bodies of Grifola from dosa. Chern.Pharm.Bull, 45: 1756-1760
Joachim Rosecke, Wilfried A. K?nig . 2000.Constituents of various wood-rotting basidiomycetes. Phytoc -hemistry, 54(6), 603~610
Kobayshi M, Hayashi T, Hayashi K. 1983. Isolatio and studies on marine Natura products(II). New polyhydroxytated Sterols from the soft coral lobolytum pauciftorum(Ehrenbery). Chem-Pharm Bull, 31, 18-48.
Koji O, Yosulori Y, Noriko M, Masao K. 1996. Sterol Constituents from the Selerotium of Polyporus umbellate Fries. Naturalm edicines, 50( 2): 170—181
Lu W., Adachi I, Kana K, Yasuta A, Toriizuka K, Ueno M. 1985. Chem. Pharm. Bull, 33, 5083
Manohar B, Basappa C, Rao D N, Divakar S. 1998. Response surface studies on cholesterol reduction in egg yolk usingβ-cy-clodextrin. Zeitschrift fuer Lebensmittel-Untersuchung und-Forschung A, 206 (3) : 189–192.
Robert O, WilliamsⅢ, Vompann M, Mongkol S. 1998. Characterizafion of an inclusion complex of β-cycledex-trin and cholesterol by aeration at elevated temperatures. Biochemical Engineering Journal, 22(3): 197-205.
Seoung wook jun.min-soo kim.jeong-soo kim.2007.Perhapation and characterization of simvastafin/hydroxypropy l-β-cyclodextrin inclusion complex using supercritical anti-solvent(SAS) process
Shinjiro Y, Hajime K, Talmaki M, Xin H X, Hajime U. 2005. Cholesterol recovery from inclusion complex T. ohsawa, M. yukawa, C. takao, M. nrayama, AND H. eando. 1992. Studies on constituents of Fruit Body of Polyporus umbellatus Fries and Their cytotoxic Activity. chem-pharm.Bull. 40 (1): 143-147
Takatomi Ohsawa, et al. 1992, Chem Pharm Bull, 40(1), 143
Tayade.Pralhad.Kale.Rajen 2004 drakumar Study of freeze-dried quercefin cyclodextrin binary systems by DSC,FT-IR,X-ray diffraction and SEM analysis
Weete JD. 1989, Structure and function sterols in fungi . Adv Lipid Res, 23: 1152 167 .
XU Jin-tang(徐锦堂). 1997.Chinese Medicinal Fungus(中国药用真菌).Peking Union Medical College and Beijing Medical Univercity Press(北京医科大学.中国协和医科大学联合出版), 150
Yasunori Inaoka. 1994, Chem Pharm Bull, 42(3), 530
You J S, Hau D M, Chen K T, Huang H F.1994.Combined effects of chuling (Polyporus umbellatus) extract and mitomycin C on experimental liver caheer. Am. J. Chin. Med., 22 (1): 19-28.