知母提取物抗真菌作用实验研究
摘要
由真菌引起的皮肤癣菌病发病率很高,且流行广泛,难以治愈,是目前较难控制的皮肤病之一。临床上常用治疗皮肤癣菌病的药物多为化学合成药,虽然都有肯定的疗效,但多数药物价格昂贵,又有一定的副作用。因此,寻找和开发新的高效、低毒抗真菌药物成为人们研究的热点。
从天然植物中提取筛选抗真菌活性成分,进而开发利用是一个重要的研究方向,具有广阔的前景。有研究报导中药知母煎剂对常见皮肤癣菌有抑制作用,但有关此方面研究资料甚少,利用真菌感染动物模型研究体内实验的未见报道。本实验通过液体试管法研究了知母提取物的体外抗真菌活性;建立豚鼠须癣毛癣菌感染体癣模型,以达克宁为对照,观察知母提取物在实验动物上的抗真菌作用;通过急性毒性实验、皮肤刺激实验、皮肤过敏实验,观察知母提取物乳膏及乳膏基质进行皮肤给药时对动物皮肤的毒副作用。
实验结果表明,知母提取物对须癣毛癣菌,石膏样小孢子菌,絮状表皮癣菌和犬小孢子菌均有较强的抑制作用,对白色念珠菌和新生隐球菌也有一定的抑制作用;治疗效果实验中,知母提取物乳膏具有抗须癣毛癣菌感染作用,其疗效与达克宁乳膏比较无显著差异。毒理学实验表明,知母提取物乳膏对皮肤无毒副作用,无刺激性。
Objective:
Dermatomycosis which is caused by superificial fungi is a common,frequently-occurring disease in dermatology. It is one of the most difficultcontrol dermatoses, at present. Since the first artificially antifungal drugAmphotericin B was synthesized in 60's, human has been fighting with thefungi disease for more than 40 years. Now the antifungal drugs include manypolyene Antibiotics, nonpolyene Antibiotics and chemosynthesis medicine.Though they have definite effect, most of them are expensive, with great sideeffect, drug resistant strain appeared fast and highly relapse. The defection ofthese drugs are still bothering patients and medical researchers and forcingmedical researchers to find the broad-spectrum ,high performance and lowpriced antifungal drugs continuously, at the same time. The traditionalChinese medicine has a long history in treating mycosis .In the past severaldecades, people have been done a lot of work for antifungal drugs intraditional Chinese medicine, and got a lot of progress already. TraditionalChinese medicine has many advantages with high performance, less sideeffect, low drug resistant strain and low price, so it is a bright orientation toselect the antifungal drugs from herbal medicines and to develop. There aresome reports about the Anemarrhena asphodeloides Bge decoction has theantifungal activity. And there is no report about establishing the animal modelby fungi. The aim for this experiment is to determine the antifungal effectualparts of Anemarrhena asphodeloides Bge, to test if the extraction has the
antifungal activity in vitro, to observe the therapeutic effect of the guinea pigtinea corporis model in comparing Anemarrhena asphodeloides Bge extractioncream with Miconzole Nitrate cream, to observe the dermal toxic effectbetween Anemarrhena asphodeloides Bge extraction cream and cream basematerial by using on skin.Method:1. Detecting the antifungal effect of Anemarrhena asphodeloides Bgeextraction to Candidaalbicans, Cryptococcus neoformans, Trichophytonmentagrophytes, Epidermophyton floccosum,Microsporum canis andMicrosporum gypseum by method of liquid test tube.2. Establishing guinea pig tinea corporis model by Trichophytonmentagrophytes, observing the anti-fungi function of Anemarrhenaasphodeloides Bge extraction cream to the experimental animal model, theMiconazole Nitrate cream as positive group.3. toxicology experiments.Appraising the security of Anemarrhena asphodeloides Bge extractionthrough the acute toxic experiment, skin stimulation experiment, skinsensitization experiment .Results:1. In vitro anti-fungi experimentThe extraction of Anemarrhena asphodeloides Bge has inhibitory activityon 6 kinds of experimental fungi. The extraction has the activity of inhibitionand killing to Trichophyton mentagrophytes、Epidermophyton floccosum、Microsporum canis、Microsporum gypseum ,especially to Trichophytonmentagrophytes.
2. Evaluating the therapeutic effect of Anemarrhena asphodeloides Bgeextraction cream to the guinea pig tinea corporis model.The two weeks cure rate of Anemarrhena asphodeloides Bge extractioncream and Miconazole Nitrate cream is respectively 85 percent and 80percent Both of them don't have any significance difference (P> 0.05 ) bystatistical analysis.3. Acute toxicity testWe don't determine the LD50 value, but we conclude that the maximaltoreance dose value of mice is more than 15g/kg by the acute toxicity test.According to acute toxicity appraising results, Anemarrhena asphodeloidesBge extraction has no toxicity.4. Skin stimulating experimentAnemarrhena asphodeloides Bge extraction cream and cream don't haveany obvious difference between normal skin and damaged skin, according tothe result of the experiment, we can conclude that both Anemarrhenaasphodeloides Bge extraction cream and cream don't have skin stimulationeffect.5. Skin allergic experiment2, 4-dinitrochlorobenzene, as positive medicine group, is given theexperimental animals' skin, the skin appears obvious slight erythema, noedema and all experimental animals have cutaneous anaphylaxis. After 6hours, cream as negative medicine group, has slightly erythema only oneexperimental animal, but disappeared after 48 hours and no cutaneousanaphylaxis. Anemarrhena asphodeloides Bge extraction cream group animal'sskin doesn't have any erythema and edema within 72 hours, have no
difference with the blank control group. So we can sure that Anemarrhenaasphodeloides Bge extraction can't bring allergy.Conclusion:1. Anemarrhena asphodeloides Bge extraction has certain inhibition effect toCandida albicans、Cryptococcus neoformans and killing activity toTrichophyton mentagrophytes、Epidermophyton floccosum、Microsporumcanis、Microsporum gypseum etc.2. Anemarrhena asphodeloides Bge extraction cream and MiconazoleNitrate cream have no obvious difference at therapeutic effect to guineapig skin tinea corporis model.3. Anemarrhena asphodeloides Bge extraction cream has no toxicity andside effect of animal's skin, it can be used safely.
从天然植物中提取筛选抗真菌活性成分,进而开发利用是一个重要的研究方向,具有广阔的前景。有研究报导中药知母煎剂对常见皮肤癣菌有抑制作用,但有关此方面研究资料甚少,利用真菌感染动物模型研究体内实验的未见报道。本实验通过液体试管法研究了知母提取物的体外抗真菌活性;建立豚鼠须癣毛癣菌感染体癣模型,以达克宁为对照,观察知母提取物在实验动物上的抗真菌作用;通过急性毒性实验、皮肤刺激实验、皮肤过敏实验,观察知母提取物乳膏及乳膏基质进行皮肤给药时对动物皮肤的毒副作用。
实验结果表明,知母提取物对须癣毛癣菌,石膏样小孢子菌,絮状表皮癣菌和犬小孢子菌均有较强的抑制作用,对白色念珠菌和新生隐球菌也有一定的抑制作用;治疗效果实验中,知母提取物乳膏具有抗须癣毛癣菌感染作用,其疗效与达克宁乳膏比较无显著差异。毒理学实验表明,知母提取物乳膏对皮肤无毒副作用,无刺激性。
Objective:
Dermatomycosis which is caused by superificial fungi is a common,frequently-occurring disease in dermatology. It is one of the most difficultcontrol dermatoses, at present. Since the first artificially antifungal drugAmphotericin B was synthesized in 60's, human has been fighting with thefungi disease for more than 40 years. Now the antifungal drugs include manypolyene Antibiotics, nonpolyene Antibiotics and chemosynthesis medicine.Though they have definite effect, most of them are expensive, with great sideeffect, drug resistant strain appeared fast and highly relapse. The defection ofthese drugs are still bothering patients and medical researchers and forcingmedical researchers to find the broad-spectrum ,high performance and lowpriced antifungal drugs continuously, at the same time. The traditionalChinese medicine has a long history in treating mycosis .In the past severaldecades, people have been done a lot of work for antifungal drugs intraditional Chinese medicine, and got a lot of progress already. TraditionalChinese medicine has many advantages with high performance, less sideeffect, low drug resistant strain and low price, so it is a bright orientation toselect the antifungal drugs from herbal medicines and to develop. There aresome reports about the Anemarrhena asphodeloides Bge decoction has theantifungal activity. And there is no report about establishing the animal modelby fungi. The aim for this experiment is to determine the antifungal effectualparts of Anemarrhena asphodeloides Bge, to test if the extraction has the
antifungal activity in vitro, to observe the therapeutic effect of the guinea pigtinea corporis model in comparing Anemarrhena asphodeloides Bge extractioncream with Miconzole Nitrate cream, to observe the dermal toxic effectbetween Anemarrhena asphodeloides Bge extraction cream and cream basematerial by using on skin.Method:1. Detecting the antifungal effect of Anemarrhena asphodeloides Bgeextraction to Candidaalbicans, Cryptococcus neoformans, Trichophytonmentagrophytes, Epidermophyton floccosum,Microsporum canis andMicrosporum gypseum by method of liquid test tube.2. Establishing guinea pig tinea corporis model by Trichophytonmentagrophytes, observing the anti-fungi function of Anemarrhenaasphodeloides Bge extraction cream to the experimental animal model, theMiconazole Nitrate cream as positive group.3. toxicology experiments.Appraising the security of Anemarrhena asphodeloides Bge extractionthrough the acute toxic experiment, skin stimulation experiment, skinsensitization experiment .Results:1. In vitro anti-fungi experimentThe extraction of Anemarrhena asphodeloides Bge has inhibitory activityon 6 kinds of experimental fungi. The extraction has the activity of inhibitionand killing to Trichophyton mentagrophytes、Epidermophyton floccosum、Microsporum canis、Microsporum gypseum ,especially to Trichophytonmentagrophytes.
2. Evaluating the therapeutic effect of Anemarrhena asphodeloides Bgeextraction cream to the guinea pig tinea corporis model.The two weeks cure rate of Anemarrhena asphodeloides Bge extractioncream and Miconazole Nitrate cream is respectively 85 percent and 80percent Both of them don't have any significance difference (P> 0.05 ) bystatistical analysis.3. Acute toxicity testWe don't determine the LD50 value, but we conclude that the maximaltoreance dose value of mice is more than 15g/kg by the acute toxicity test.According to acute toxicity appraising results, Anemarrhena asphodeloidesBge extraction has no toxicity.4. Skin stimulating experimentAnemarrhena asphodeloides Bge extraction cream and cream don't haveany obvious difference between normal skin and damaged skin, according tothe result of the experiment, we can conclude that both Anemarrhenaasphodeloides Bge extraction cream and cream don't have skin stimulationeffect.5. Skin allergic experiment2, 4-dinitrochlorobenzene, as positive medicine group, is given theexperimental animals' skin, the skin appears obvious slight erythema, noedema and all experimental animals have cutaneous anaphylaxis. After 6hours, cream as negative medicine group, has slightly erythema only oneexperimental animal, but disappeared after 48 hours and no cutaneousanaphylaxis. Anemarrhena asphodeloides Bge extraction cream group animal'sskin doesn't have any erythema and edema within 72 hours, have no
difference with the blank control group. So we can sure that Anemarrhenaasphodeloides Bge extraction can't bring allergy.Conclusion:1. Anemarrhena asphodeloides Bge extraction has certain inhibition effect toCandida albicans、Cryptococcus neoformans and killing activity toTrichophyton mentagrophytes、Epidermophyton floccosum、Microsporumcanis、Microsporum gypseum etc.2. Anemarrhena asphodeloides Bge extraction cream and MiconazoleNitrate cream have no obvious difference at therapeutic effect to guineapig skin tinea corporis model.3. Anemarrhena asphodeloides Bge extraction cream has no toxicity andside effect of animal's skin, it can be used safely.
引文
[1] 中华人民共和国药典委员会.中华人民共和国药典[M].北京:人民卫生出版社,1990.184.
[2] 新文丰出版公司.新编中药大辞典(中册)台湾.新文丰出版社,1980.1076.
[3] 林启寿.中药成分化学[M].北京:科学出版社,1977.484.
[4] 边际,徐绥绪.知母化学及药理研究进展 [J].沈阳药学院学报,1993,10(2):141~144.
[5] Dong JX, Han GY,Studies on the active constituents of Anemarrhena asphodeloides bunge.Yao Xue Xue Bao. 1992,27(1):26~32.
[6] 边际, 徐绥绪,黄松,等.知母化学成分的研究[J].沈阳药科大学学报,1996,13(1):34~40.
[7] 王浴生,邓文龙,薛春生.中药药理与应用(第二版)[M].北京:人民卫生出版社,1998.709.
[8] 孟志云, 徐绥绪.知母中三个新的呋甾皂苷[J].沈阳药科大学学报,1998,15(2):130~131.
[9] 马百平,董俊兴,王秉,等.知母中呋甾皂甙的研究 [J].药学学报,1996,31(4):271~277.
[10] 孟志云, 徐绥绪,周晓棉.知母中新的甾体皂苷[J]. 沈阳药科大学学报,1998,15(4):254~256.
[11] 孟 志 云 , 徐 绥 绪 , 孟 令 宏 . 知 母 皂 甙 E1 和 E2[J]. 药 学 学报,1998,33(9):693~696.
[12] 孟 志 云 , 李 文 , 徐 绥 绪 , 等 . 知 母 的 皂 苷 成 分 [J]. 药 学 学报,1999,34(6):451~453.
[13] 孟志云, 徐绥绪,李文,等.知母中新的皂苷成分[J].中国药物化学杂志,1999,9(4):294~298.
[14] 孟 志 云 , 徐 绥 绪 . 知 母 中 的 皂 苷 成 分 [J]. 中 国 药 物 化 学 杂志,1998,8(2):135~140.
[15] 洪永福,张广明.西陵知母中甾体皂苷的分离与鉴定[J].药学学报,1999,34(7):518~512.
[16] 滕辉,郭顺星.知母须根与根茎化学成分的对比分析[J].中国中药杂志,1990,15(9):14~16.
[17] 洪永福,韩公羽,周卓伦,等.不同采集期知母根茎中芒果甙含量变化的初步研究[J].中草药,1985,16(4):31~33.
[18] Hong YF, Han GY, Guo XM. Isolation and structure determination of xanthone glycosides of Anemarrhena asphodeloides,Yao Xue Xue Bao. 1997,32(6):473~475.
[19] 狄凯军,章静波.国内外淫羊藿苷药理作用研究要览[J].自然杂志,2003,25(4):191.
[20] 刘昆兆.中药知母化学成分的研究[J].阜阳师范学院学报,1991,15:5~6.
[21] 王浴生,邓文龙等主编.中药药理与应用[M].北京:人民卫生出版社,1983.679~709.
[22] 廖洪利,吴秋业,叶光明,等.芒果苷药理研究进展[J].天津药学,2005,17(2):50~52.
[23] 李沙,甄宏,蒋杰,等.知母宁的体外抗单纯病毒疱疹病毒 II 型作用[J].华中科技大学学报(医学版)2005,34(3):304~307.
[24] Iida Y,Oh K,Saito M,et al.Planta Med,2000,66(5):145~150.
[25] 陈万生,韩军,李力,等.知母总多糖的抗炎作用[J].第二军医大学学报[J],1999,20(10):758~760.
[26] 王靖,葛盛芳,陈琦,等.知母多糖降血糖活性研究[J].中草药,1996,27(10):605~606.
[27] Miura T,Ichiki H,Iwamoto N,et al.Phytomedicine,2001,8(2):85~87.
[28] Zhang J,Meng Z, Zhang M, et al.Clin Chim Acta,1999,289(1/2):79~88.
[29] Takeda Y,Togashi H,Matsuo T,et al. J Gastroenterol,2001,36(2):79~90.
[30] 黄华艺,农朝赞,郭凌霄,等.芒果甙对肝癌细胞增殖的抑制和凋亡的诱导[J].中华消化杂志,2002,22(6):341~343.
[31] 彭志刚,罗军,夏凌辉,等.芒果甙对白血病 K562 细胞增殖抑制作用的研究[J].广西医科大学学报,2004,21(2):168~170.
[32] 马玉奎,李莉,刘国宾.知母皂苷对三氯化铝致老年性痴呆模型大鼠的作用[J].齐鲁药事,2005,24/(10):625~626.
[33] 陈勤,胡雅儿,夏宗勤.知母皂甙元对半乳糖拟痴呆型小鼠学习记忆和脑自由基代谢的影响[J].中药药理与临床,2000,16(50):14~16.
[34] 王顺官,徐江平.知母皂苷对衰老大鼠脑乙酰胆碱受体的调节作用[J].解放军药学学报,2001,17(2):71~72.
[35] 赵树进,韩丽萍,李俭洪,等.知母皂苷及其苷元对动物模型β-肾上腺素受体的调整作用[J].中国医院药学杂志,2000,20(2):70~73.
[36] 李政,王国良,金珊,等.31种常见抗菌中药对DPPH自由基清除作用的分析[J].海洋水产研究,2005,26(1):38~40.
[37] 张红雨,王芙媛,李明,等.知母宁清除活性氧作用的研究[J].辐射研究与辐射工艺学报,1997,15(4):224~227.
[38] 朱起之,赵树进.知母皂甙对糖皮质激素副作用的影响[J].实用医学杂志,2001,17(7):583~584.
[39] 冯新港,谈佩萍,易健民,等.92种野生和栽培植物或中草药的提取物杀灭钉螺筛选试验[J].中国血吸虫病防治杂志,2002,14(6):412~417.
[40] Matsuda H,Sato N,Yamazaki M,et al.Biol Pharm Bull,2001,24(5):586~587.
[41] 王道毅,陈炼,李忌,等.知母宁对阿霉素的减毒增效作用[J].天然产物研究与开发,2000,12(4):8~11.
[42] 王凤芝,陶站华,王晓惠,等.中药知母对小鼠免疫功能的影响[J].黑龙江医药科学,2002,25(3):7.
[43] 赵惠仁,胡书群,任孝衡.知母等.40 种中药对猪晶状体醛糖还原酶的抑制作用[J].徐州医学院学报,1995,15(3):235~237.
[44] 韩丽萍,赵树进,李俭洪.大孔树脂法提取知母总皂甙[J].中药材,2004,27(4):288~289.
[45] 贾春林主编.中药药剂学[M].第1版.上海:上海科学技术出版社,1986.332.
[46] 刘水平,邬国军主编.医学微生物学实验指导[M]. 第1版.兴界图书出版公司,2002.209.
[47] 曹永兵,姜远英,王宁,等.微量液基稀释法测定化合物特苄康唑的体外抗真菌活性[J].中国抗生素杂志,2000,25(3):183~187.
[48] 中药新药研究指南[M].中华人民共和国卫生部药政管理局.179.
[49] 苗明三主编.实验动物和动物实验技术[M].第 1 版.中国中医药出版社,1997.255.
[50] 李世荫,宫振钦等.实用皮肤病学[M].青海:青海人民出版社,1978.115.
[51] 朱学俊,顾有守,沈丽玉.实用皮肤病性病治疗学[M].第 2 版.北京:北京医科大学协和医科大学联合出版社,1998.201~202.
[52] 金有豫,孙颂三.药理学[M].第五版.北京:人民卫生出版社,2001.359~362.
[53] 刘潋,常海涛,屠鹏飞.抗真菌药物的研究进展[J].中国药学杂志,2001,36(4):227~230.
[54] 李厚敏,刘伟,李若瑜.美国临床实验室标准化委员会 2003 年版产孢丝状真菌药敏试验方案简介[J].中华检验医学杂志,2005,28(1):121~123.
[2] 新文丰出版公司.新编中药大辞典(中册)台湾.新文丰出版社,1980.1076.
[3] 林启寿.中药成分化学[M].北京:科学出版社,1977.484.
[4] 边际,徐绥绪.知母化学及药理研究进展 [J].沈阳药学院学报,1993,10(2):141~144.
[5] Dong JX, Han GY,Studies on the active constituents of Anemarrhena asphodeloides bunge.Yao Xue Xue Bao. 1992,27(1):26~32.
[6] 边际, 徐绥绪,黄松,等.知母化学成分的研究[J].沈阳药科大学学报,1996,13(1):34~40.
[7] 王浴生,邓文龙,薛春生.中药药理与应用(第二版)[M].北京:人民卫生出版社,1998.709.
[8] 孟志云, 徐绥绪.知母中三个新的呋甾皂苷[J].沈阳药科大学学报,1998,15(2):130~131.
[9] 马百平,董俊兴,王秉,等.知母中呋甾皂甙的研究 [J].药学学报,1996,31(4):271~277.
[10] 孟志云, 徐绥绪,周晓棉.知母中新的甾体皂苷[J]. 沈阳药科大学学报,1998,15(4):254~256.
[11] 孟 志 云 , 徐 绥 绪 , 孟 令 宏 . 知 母 皂 甙 E1 和 E2[J]. 药 学 学报,1998,33(9):693~696.
[12] 孟 志 云 , 李 文 , 徐 绥 绪 , 等 . 知 母 的 皂 苷 成 分 [J]. 药 学 学报,1999,34(6):451~453.
[13] 孟志云, 徐绥绪,李文,等.知母中新的皂苷成分[J].中国药物化学杂志,1999,9(4):294~298.
[14] 孟 志 云 , 徐 绥 绪 . 知 母 中 的 皂 苷 成 分 [J]. 中 国 药 物 化 学 杂志,1998,8(2):135~140.
[15] 洪永福,张广明.西陵知母中甾体皂苷的分离与鉴定[J].药学学报,1999,34(7):518~512.
[16] 滕辉,郭顺星.知母须根与根茎化学成分的对比分析[J].中国中药杂志,1990,15(9):14~16.
[17] 洪永福,韩公羽,周卓伦,等.不同采集期知母根茎中芒果甙含量变化的初步研究[J].中草药,1985,16(4):31~33.
[18] Hong YF, Han GY, Guo XM. Isolation and structure determination of xanthone glycosides of Anemarrhena asphodeloides,Yao Xue Xue Bao. 1997,32(6):473~475.
[19] 狄凯军,章静波.国内外淫羊藿苷药理作用研究要览[J].自然杂志,2003,25(4):191.
[20] 刘昆兆.中药知母化学成分的研究[J].阜阳师范学院学报,1991,15:5~6.
[21] 王浴生,邓文龙等主编.中药药理与应用[M].北京:人民卫生出版社,1983.679~709.
[22] 廖洪利,吴秋业,叶光明,等.芒果苷药理研究进展[J].天津药学,2005,17(2):50~52.
[23] 李沙,甄宏,蒋杰,等.知母宁的体外抗单纯病毒疱疹病毒 II 型作用[J].华中科技大学学报(医学版)2005,34(3):304~307.
[24] Iida Y,Oh K,Saito M,et al.Planta Med,2000,66(5):145~150.
[25] 陈万生,韩军,李力,等.知母总多糖的抗炎作用[J].第二军医大学学报[J],1999,20(10):758~760.
[26] 王靖,葛盛芳,陈琦,等.知母多糖降血糖活性研究[J].中草药,1996,27(10):605~606.
[27] Miura T,Ichiki H,Iwamoto N,et al.Phytomedicine,2001,8(2):85~87.
[28] Zhang J,Meng Z, Zhang M, et al.Clin Chim Acta,1999,289(1/2):79~88.
[29] Takeda Y,Togashi H,Matsuo T,et al. J Gastroenterol,2001,36(2):79~90.
[30] 黄华艺,农朝赞,郭凌霄,等.芒果甙对肝癌细胞增殖的抑制和凋亡的诱导[J].中华消化杂志,2002,22(6):341~343.
[31] 彭志刚,罗军,夏凌辉,等.芒果甙对白血病 K562 细胞增殖抑制作用的研究[J].广西医科大学学报,2004,21(2):168~170.
[32] 马玉奎,李莉,刘国宾.知母皂苷对三氯化铝致老年性痴呆模型大鼠的作用[J].齐鲁药事,2005,24/(10):625~626.
[33] 陈勤,胡雅儿,夏宗勤.知母皂甙元对半乳糖拟痴呆型小鼠学习记忆和脑自由基代谢的影响[J].中药药理与临床,2000,16(50):14~16.
[34] 王顺官,徐江平.知母皂苷对衰老大鼠脑乙酰胆碱受体的调节作用[J].解放军药学学报,2001,17(2):71~72.
[35] 赵树进,韩丽萍,李俭洪,等.知母皂苷及其苷元对动物模型β-肾上腺素受体的调整作用[J].中国医院药学杂志,2000,20(2):70~73.
[36] 李政,王国良,金珊,等.31种常见抗菌中药对DPPH自由基清除作用的分析[J].海洋水产研究,2005,26(1):38~40.
[37] 张红雨,王芙媛,李明,等.知母宁清除活性氧作用的研究[J].辐射研究与辐射工艺学报,1997,15(4):224~227.
[38] 朱起之,赵树进.知母皂甙对糖皮质激素副作用的影响[J].实用医学杂志,2001,17(7):583~584.
[39] 冯新港,谈佩萍,易健民,等.92种野生和栽培植物或中草药的提取物杀灭钉螺筛选试验[J].中国血吸虫病防治杂志,2002,14(6):412~417.
[40] Matsuda H,Sato N,Yamazaki M,et al.Biol Pharm Bull,2001,24(5):586~587.
[41] 王道毅,陈炼,李忌,等.知母宁对阿霉素的减毒增效作用[J].天然产物研究与开发,2000,12(4):8~11.
[42] 王凤芝,陶站华,王晓惠,等.中药知母对小鼠免疫功能的影响[J].黑龙江医药科学,2002,25(3):7.
[43] 赵惠仁,胡书群,任孝衡.知母等.40 种中药对猪晶状体醛糖还原酶的抑制作用[J].徐州医学院学报,1995,15(3):235~237.
[44] 韩丽萍,赵树进,李俭洪.大孔树脂法提取知母总皂甙[J].中药材,2004,27(4):288~289.
[45] 贾春林主编.中药药剂学[M].第1版.上海:上海科学技术出版社,1986.332.
[46] 刘水平,邬国军主编.医学微生物学实验指导[M]. 第1版.兴界图书出版公司,2002.209.
[47] 曹永兵,姜远英,王宁,等.微量液基稀释法测定化合物特苄康唑的体外抗真菌活性[J].中国抗生素杂志,2000,25(3):183~187.
[48] 中药新药研究指南[M].中华人民共和国卫生部药政管理局.179.
[49] 苗明三主编.实验动物和动物实验技术[M].第 1 版.中国中医药出版社,1997.255.
[50] 李世荫,宫振钦等.实用皮肤病学[M].青海:青海人民出版社,1978.115.
[51] 朱学俊,顾有守,沈丽玉.实用皮肤病性病治疗学[M].第 2 版.北京:北京医科大学协和医科大学联合出版社,1998.201~202.
[52] 金有豫,孙颂三.药理学[M].第五版.北京:人民卫生出版社,2001.359~362.
[53] 刘潋,常海涛,屠鹏飞.抗真菌药物的研究进展[J].中国药学杂志,2001,36(4):227~230.
[54] 李厚敏,刘伟,李若瑜.美国临床实验室标准化委员会 2003 年版产孢丝状真菌药敏试验方案简介[J].中华检验医学杂志,2005,28(1):121~123.