甘草黄酮的分离鉴定、药效及其指纹图谱研究
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摘要
随着对甘草药理作用的进一步认识,全世界对甘草的需求日益增长,甘草资源急剧减少,合理开发甘草资源已十分必要。甘草的植物化学成分十分复杂,以往人们只对甘草中的三萜类化合物作了大量的研究,对其质量评价也仅以甘草酸的含量作为标准,难以全面地评价甘草的质量。近年来,对甘草药理作用的深入研究表明,甘草黄酮类化合物具有多种药理作用,是一类非常有效的活性成分。因此,本课题对甘草中总黄酮的分离纯化及重要单体甘草查尔酮A和光甘草定的分离鉴定和药效进行研究,同时建立胀果甘草黄酮类化合物的HPLC指纹图谱,为甘草黄酮资源的有效、合理开发和质量评价提供依据。
本论文首先建立了甘草中总黄酮的含量测定方法,测定胀果甘草、光果甘草和乌拉尔甘草中总黄酮平均含量分别为:4.476 %、2.478 %和1.372%。
对比研究了甘草总黄酮的几种不同提取方法,结果超声辅助法提取甘草总黄酮在提取时间和提取效率方面均优于其他几种方法。利用响应面分析法优化甘草总黄酮超声辅助提取的工艺参数,得到超声辅助提取甘草总黄酮的最佳工艺,以此工艺进行超声辅助提取甘草总黄酮一次提取率达到80.56 %。通过比较,选择AB-8大孔树脂纯化甘草总黄酮,优化了AB-8大孔树脂纯化甘草总黄酮的实验条件,经过纯化的甘草总黄酮含量达到60 %以上,收率80 %以上。
建立了基于生物信息学的甘草黄酮类化学成分抗乙酰胆碱酯酶活性虚拟评价的新方法,在光果甘草几种主要化合物成分中,光甘草定与乙酰胆碱酯酶的对接能和结合能均较小,结合“吻合度”很好,与三种乙酰胆碱酯酶抑制剂(加兰他敏、石杉碱甲和美曲膦脂)的对接能和结合能接近。通过分子对接分析,将光甘草定A环或B环上的氢原子用氟原子取代进行对接分析,得到对接能和结合能与光甘草定相似的小分子5’-氟光甘草定。
根据分子对接结果,从光果甘草中提取分离并鉴定了光甘草定,还建立了甘草中光甘草定的含量测定方法,测定了光果甘草中光甘草定的平均含量为0.199 %,而胀果甘草和乌拉尔甘草中没有检测到光甘草定。
以东莨宕碱所致的小鼠认知功能障碍为模型,研究了光甘草定对小鼠学习记忆功能的影响,在“Y”型电迷宫实验和跳台实验中,光甘草定组(2, 4 mg/kg)与模型组相比,学习记忆成绩均有显著提高,且光甘草定大剂量组(4 mg/kg)获得了与阳性组脑复康(400 mg/kg)相当的结果。中高剂量的光甘草定(2,4 mg/kg)和阳性药美曲膦脂(50 mg/kg)能使小鼠大脑乙酰胆碱酯酶的活性分别下降14.4%, 19.0%和21.3%。
本论文还从胀果甘草中提取分离了甘草查尔酮A,并运用LC-MS和NMR对其结构进行了鉴定。并建立了甘草中甘草查尔酮A的含量测定方法,测定了胀果甘草和光果甘草中甘草查尔酮A的平均含量分别为0.411 %和0.054 %,而乌拉尔甘草中没有检测到甘草查尔酮A。
以二甲苯所致的小鼠耳肿胀和角叉菜胶所致的大鼠足肿胀研究了甘草查尔酮A的抗炎效果,结果表明甘草查尔酮A (5,10 mg/kg)能显著抑制二甲苯所致的小鼠耳肿胀。三个剂量2.5,5和10 mg/kg的甘草查尔酮A均能显著抑制角叉菜胶所致的大鼠足肿胀。甘草查尔酮A (0.1, 0.5和1μg/mL )能显著抑制COX-2的合成和活性,且高剂量(1μg/mL)的甘草查尔酮A对COX-2合成的抑制作用要强于0.5μg/mL的阳性药吲哚美辛。
采用HPLC方法,以乙腈-0.1%冰醋酸为流动相进行梯度洗脱,对10批胀果甘草样品中黄酮类化合物进行了分析研究,并采用计算机软件对色谱峰数据进行评价,建立了胀果甘草黄酮类成分的HPLC指纹图谱,可利用其特征峰有效的评价甘草药材的质量。
The demands for licorice in the world are growing with the further research on pharmacological effects of licorice. Thus, it is necessary to exploit rationally resources of licorice due to its sharp decline. The phytochemical components of licorice are so complex. However, only the terpenoids such as glycyrrhizic acid were investigated in-depth previously. It is not reasonable to assess the quality of licorice in general only by referring to content of glycyrrhizic acid. In recent years, the in-depth study of pharmacological effects of licorice indicated that flavonoids from licorice were effective active ingredients, possessing a variety of pharmacological effects, and have become research hotspots in herbs. In this study, the isolation, identification and pharmacodynamics of total flavonoids of licorice (TFL), licochalcone A and glabridin were investigated. Meanwhile, the HPLC fingerprint of flavonoids of Glycyrrhiza inflata was established for the effective and reasonable exploitation and evaluation of flavonoids of licorice.
The method for the determination of TFL was established. The concents of TFL in Glycyrrhiza inflata Bat., Glycyrrhiza glabra L. and Glycyrrhiza uralensis Fisch were 4.476 %, 2.478 % and 1.372 %, respectively..
The extraction techniques for TFL were investigated by contrast. The results showed that ultrasonic-assisted extraction was better than other extractions. The extracting parameters of TFL through ultrasonic-assisted extraction were determined using Response Surface Methododlogy. The extraction of TFL was 80.56 % under the obtained better process conditions. The optimum purification conditions of TFL with macroporous resin AB-8 was obtained. With macroporous resin to adsorb and purify, the yield of TFL was higher than 80 % and the content was higher than 60 %.
Based on numerator docking, a new method was established for evaluating the anti-acetylcholinesterase activity of flavonoids from licorice. The results showed that the docking and binding energy of glabridin were lower than those of others components, and were considerable with those of acetylcholinesterase inhibitors (galanthamine, huperzine and metrifonate). The formed compounds are steady. Based on molecular docking analysis, 5’-fluglabridin was obtained with equivalent energy with glabridin when replacing H atom or hydroxy in the A-ring or B-ring of glabridin with F atom.
According to molecular docking evaluation, glabridin was isolated from Glycyrrhiza glabra and its structure was identified. The method for the determination of glabridin was established. The content of glabridin in Glycyrrhiza glabra L. was 0.199 %, but it could not be detected in Glycyrrhiza inflata Bat. or Glycyrrhiza uralensis Fisch.
The effects of glabridin on scopolamine-induced learning and memory impairment were studied in mice. In Y-electronic maze test and step-down test, glabridin (2, 4 mg kg-1, p.o.) could significantly enhance the learning and memory ability of mice compared with the model group in mice. The effect exhibited by glabridin at higher dose (4 mg/kg) was considerable with that of piracetam (400 mg kg-1, i.p.), a clinically used nootropic agent. Furthermore, both glabridin (2, 4 mg kg-1) and metrifonate (50 mg kg-1, i.p.), used as a standard drug, can reduced the brain cholinesterase activity to 14.4 %, 19.0 % and 21.3 %, respectively, compared with control group.
Licochalcone A was isolated from the roots of Glycyrrhiza inflata and identified with LC-MS and NMR. The method for determination of licochalcone A was established. The contents of licochalcone A in Glycyrrhiza inflata Bat. and Glycyrrhiza glabra L. were 0.411 % and 0.054 %, respectively, but it could not be detected in Glycyrrhiza uralensis Fisch.
Furthermore, the anti-inflammatory activity of licochalcone A was evaluated in xylene-induced mice ear edema and carrageenan-induced paw edema tests. At the concentrations of 5 and 10 mg/ear, licochalcone A showed remarkable effect against acute inflammation induced by xylene, and at the doses of 2.5, 5, 10 mg/kg (p.o.), licochalcone A reduced significantly paw edema induced by carrageenan compared with the control group. Both COX-2 activity and expression were significantly inhibited by licochalcone A at all tested doses. The inhibitory effect of glabridin on COX-2 expression exhibited by glabridin at higher dose (1μg/mL) was higher than that of indomethacin (0.5μg mL-1).
The flavonoids from 10 samples of Glycyrrhiza inflata were analysized with HPLC using a step gradient of acetonitrile-0.1 % acetic acid. The HPLC fingerprint chromatogram of flavonoids from Glycyrrhiza inflata was established using computer software to evaluate the data of chromatographic peaks. It is effective to evaluate the quality of licorice in virtue of the characteristic peaks of HPLC fingerprint chromatogram
本论文首先建立了甘草中总黄酮的含量测定方法,测定胀果甘草、光果甘草和乌拉尔甘草中总黄酮平均含量分别为:4.476 %、2.478 %和1.372%。
对比研究了甘草总黄酮的几种不同提取方法,结果超声辅助法提取甘草总黄酮在提取时间和提取效率方面均优于其他几种方法。利用响应面分析法优化甘草总黄酮超声辅助提取的工艺参数,得到超声辅助提取甘草总黄酮的最佳工艺,以此工艺进行超声辅助提取甘草总黄酮一次提取率达到80.56 %。通过比较,选择AB-8大孔树脂纯化甘草总黄酮,优化了AB-8大孔树脂纯化甘草总黄酮的实验条件,经过纯化的甘草总黄酮含量达到60 %以上,收率80 %以上。
建立了基于生物信息学的甘草黄酮类化学成分抗乙酰胆碱酯酶活性虚拟评价的新方法,在光果甘草几种主要化合物成分中,光甘草定与乙酰胆碱酯酶的对接能和结合能均较小,结合“吻合度”很好,与三种乙酰胆碱酯酶抑制剂(加兰他敏、石杉碱甲和美曲膦脂)的对接能和结合能接近。通过分子对接分析,将光甘草定A环或B环上的氢原子用氟原子取代进行对接分析,得到对接能和结合能与光甘草定相似的小分子5’-氟光甘草定。
根据分子对接结果,从光果甘草中提取分离并鉴定了光甘草定,还建立了甘草中光甘草定的含量测定方法,测定了光果甘草中光甘草定的平均含量为0.199 %,而胀果甘草和乌拉尔甘草中没有检测到光甘草定。
以东莨宕碱所致的小鼠认知功能障碍为模型,研究了光甘草定对小鼠学习记忆功能的影响,在“Y”型电迷宫实验和跳台实验中,光甘草定组(2, 4 mg/kg)与模型组相比,学习记忆成绩均有显著提高,且光甘草定大剂量组(4 mg/kg)获得了与阳性组脑复康(400 mg/kg)相当的结果。中高剂量的光甘草定(2,4 mg/kg)和阳性药美曲膦脂(50 mg/kg)能使小鼠大脑乙酰胆碱酯酶的活性分别下降14.4%, 19.0%和21.3%。
本论文还从胀果甘草中提取分离了甘草查尔酮A,并运用LC-MS和NMR对其结构进行了鉴定。并建立了甘草中甘草查尔酮A的含量测定方法,测定了胀果甘草和光果甘草中甘草查尔酮A的平均含量分别为0.411 %和0.054 %,而乌拉尔甘草中没有检测到甘草查尔酮A。
以二甲苯所致的小鼠耳肿胀和角叉菜胶所致的大鼠足肿胀研究了甘草查尔酮A的抗炎效果,结果表明甘草查尔酮A (5,10 mg/kg)能显著抑制二甲苯所致的小鼠耳肿胀。三个剂量2.5,5和10 mg/kg的甘草查尔酮A均能显著抑制角叉菜胶所致的大鼠足肿胀。甘草查尔酮A (0.1, 0.5和1μg/mL )能显著抑制COX-2的合成和活性,且高剂量(1μg/mL)的甘草查尔酮A对COX-2合成的抑制作用要强于0.5μg/mL的阳性药吲哚美辛。
采用HPLC方法,以乙腈-0.1%冰醋酸为流动相进行梯度洗脱,对10批胀果甘草样品中黄酮类化合物进行了分析研究,并采用计算机软件对色谱峰数据进行评价,建立了胀果甘草黄酮类成分的HPLC指纹图谱,可利用其特征峰有效的评价甘草药材的质量。
The demands for licorice in the world are growing with the further research on pharmacological effects of licorice. Thus, it is necessary to exploit rationally resources of licorice due to its sharp decline. The phytochemical components of licorice are so complex. However, only the terpenoids such as glycyrrhizic acid were investigated in-depth previously. It is not reasonable to assess the quality of licorice in general only by referring to content of glycyrrhizic acid. In recent years, the in-depth study of pharmacological effects of licorice indicated that flavonoids from licorice were effective active ingredients, possessing a variety of pharmacological effects, and have become research hotspots in herbs. In this study, the isolation, identification and pharmacodynamics of total flavonoids of licorice (TFL), licochalcone A and glabridin were investigated. Meanwhile, the HPLC fingerprint of flavonoids of Glycyrrhiza inflata was established for the effective and reasonable exploitation and evaluation of flavonoids of licorice.
The method for the determination of TFL was established. The concents of TFL in Glycyrrhiza inflata Bat., Glycyrrhiza glabra L. and Glycyrrhiza uralensis Fisch were 4.476 %, 2.478 % and 1.372 %, respectively..
The extraction techniques for TFL were investigated by contrast. The results showed that ultrasonic-assisted extraction was better than other extractions. The extracting parameters of TFL through ultrasonic-assisted extraction were determined using Response Surface Methododlogy. The extraction of TFL was 80.56 % under the obtained better process conditions. The optimum purification conditions of TFL with macroporous resin AB-8 was obtained. With macroporous resin to adsorb and purify, the yield of TFL was higher than 80 % and the content was higher than 60 %.
Based on numerator docking, a new method was established for evaluating the anti-acetylcholinesterase activity of flavonoids from licorice. The results showed that the docking and binding energy of glabridin were lower than those of others components, and were considerable with those of acetylcholinesterase inhibitors (galanthamine, huperzine and metrifonate). The formed compounds are steady. Based on molecular docking analysis, 5’-fluglabridin was obtained with equivalent energy with glabridin when replacing H atom or hydroxy in the A-ring or B-ring of glabridin with F atom.
According to molecular docking evaluation, glabridin was isolated from Glycyrrhiza glabra and its structure was identified. The method for the determination of glabridin was established. The content of glabridin in Glycyrrhiza glabra L. was 0.199 %, but it could not be detected in Glycyrrhiza inflata Bat. or Glycyrrhiza uralensis Fisch.
The effects of glabridin on scopolamine-induced learning and memory impairment were studied in mice. In Y-electronic maze test and step-down test, glabridin (2, 4 mg kg-1, p.o.) could significantly enhance the learning and memory ability of mice compared with the model group in mice. The effect exhibited by glabridin at higher dose (4 mg/kg) was considerable with that of piracetam (400 mg kg-1, i.p.), a clinically used nootropic agent. Furthermore, both glabridin (2, 4 mg kg-1) and metrifonate (50 mg kg-1, i.p.), used as a standard drug, can reduced the brain cholinesterase activity to 14.4 %, 19.0 % and 21.3 %, respectively, compared with control group.
Licochalcone A was isolated from the roots of Glycyrrhiza inflata and identified with LC-MS and NMR. The method for determination of licochalcone A was established. The contents of licochalcone A in Glycyrrhiza inflata Bat. and Glycyrrhiza glabra L. were 0.411 % and 0.054 %, respectively, but it could not be detected in Glycyrrhiza uralensis Fisch.
Furthermore, the anti-inflammatory activity of licochalcone A was evaluated in xylene-induced mice ear edema and carrageenan-induced paw edema tests. At the concentrations of 5 and 10 mg/ear, licochalcone A showed remarkable effect against acute inflammation induced by xylene, and at the doses of 2.5, 5, 10 mg/kg (p.o.), licochalcone A reduced significantly paw edema induced by carrageenan compared with the control group. Both COX-2 activity and expression were significantly inhibited by licochalcone A at all tested doses. The inhibitory effect of glabridin on COX-2 expression exhibited by glabridin at higher dose (1μg/mL) was higher than that of indomethacin (0.5μg mL-1).
The flavonoids from 10 samples of Glycyrrhiza inflata were analysized with HPLC using a step gradient of acetonitrile-0.1 % acetic acid. The HPLC fingerprint chromatogram of flavonoids from Glycyrrhiza inflata was established using computer software to evaluate the data of chromatographic peaks. It is effective to evaluate the quality of licorice in virtue of the characteristic peaks of HPLC fingerprint chromatogram
引文
[1]谷会岩.中国甘草资源生态学研究[D].东北林业大学, 2001.
[2]郑虎占,董泽宏,余靖.中药现代研究与应用[M].北京:学苑出版社, 1997, 1256-1279.
[3]周成明. 80种常用中草药栽培[M].第二版;北京:中国农业出版社, 2002, 1-4.
[4] Ichimura, M, Furuno, T, Takahashi, T, et al. Enzymic O-methylation of isoliquiritigenin and licodione in alfalfa and licorice culture [J]. Phytochemistry. 1997, 44(6): 991-995.
[5]杜等.甘草毛状根培养系统的建立及化学成分分析[J].植物资源与环境学报. 2001, 10(1): 7-10.
[6]郭江平.乌拉尔甘草大面积人工种植试验初报[J].干旱地区农业研究. 2000, 18(4): 133-136.
[7]于林清,何茂泰,王照兰.甘草组织培养快速繁殖技术研究[J].中国草地. 1999, 18(1): 12-14.
[8]殷生章.一个新的二氢黄酮苷[J].中国药科大学学报. 1999, 30(1): 19-20.
[9] Hatano, T, Shintani, Y, Aga, Y. Phenolics constituents of licoriceⅧ. Structures of glicophenone and glicoisoflavanone and effects of licorice phenolics on methicillin-resistant stanphlococcus aureus [J]. Chem Pharm Bull. 2000, 48(9): 1286-1292.
[10] Hatano, T, Takagi, M, Ito, H. Phenolic constituents of liquoriceⅦ. A new chalcone with a potent radical activity and accompanying phenolics from liquorice [J]. Chem Pharm Bull. 1997, 45(9): 1485-1492.
[11] Shibano, M, Matsumoto, Y, Nakao, E. Studies on the index compounds for HPLC analysis of Glycyrrhiza uralensis [J]. Heterocycles. 1997, 45(10): 2053-2060.
[12] Kinoshita, T, Kajiyama, K, Hiraga, Y. Isoflavan derivaties from Glycyrrhizaglabra [J]. Heterocycles. 1996, 43(3): 581-587.
[13] Fukai, T, Tantai, L, Nomura, T. Isoprenoid-substituted flavonoids from Glycyrrhiza glabra [J]. Heterocycles. 1996, 43(2): 531-532.
[14] Fukai, T, Cai, B, Maruno, K. "Phenolic costituents of Glycyrrhiza species" An isoprenylated flavanone from Glycyrrhiza glabra and recassay of licorice phenols [J]. Phytochemistry. 1998, 49(7): 2005-2013.
[15] Asada, Y, Li, W, Yoshikawa. The first prenylated biaurone, licoagrone from hairy root cultures of Glycyrrhiza glabra [J]. Phytochemistry. 1999, 50(6): 1015-1019.
[16] Li, W, Asada, Y, Yoshikawa, T. Antimicrobial flavonoids from Glycyrrhiza glabra hairy root culture [J]. Planta Med. 1998, 64(8): 746-747.
[17]王邠,邹坤.胀果甘草中2个新的二氢黄酮甙[J].药学学报. 1997, 32(3): 199-202.
[18] Fukai, T, Nomura, T. Isoprenoid-substituted flavonoids from roots of Glycyrrhiza inflata [J]. Phytochemistry. 1995, 38(3): 759-765.
[19]李伟东,阚毓铭.刺果甘草化学成分的研究[J].中国药学杂志. 1999, 34(1): 11-14.
[20]张聿梅,许旭东,胡碧煌.黄甘草异黄酮成分的研究[J].药学学报. 1997, 32(4): 301-304.
[21] Kitagawa, I, Chen, W, Hori, K. Chemical studies of chinese licorice-rootsⅡ. Five new flavonoid constituents from the roots of Glycyrrhiza aspera pall collected in xinjiang [J]. Chem Pharm Bull. 1998, 46(10): 1511-1517.
[22] Fuhrman, B, Volkova, N. Antiatherosclerotic effects of licorice extracts supplementation on hypercholesterolemic patients: increased resistance of LDL to atherogenic modifications, reduced plasma plasma lipid levels, and decreased systolic blood pressure. [J]. Nutrition. 2002, 18: 268-273.
[23] Vaya, J, Belinky, P A, Aviram, M. Antioxidant constituents from licorice roots: isolation, structure elucidation and antioxidative capacity toward LDL oxidation [J]. Free Rad Biol Med. 1997, 23(2): 302-313.
[24] HaraguchiH;ishikawaH, MizutaniK;TamuraY;kinoshitaT. Antioxidative andsuperoxide scavenging activities of retrochalcones in Glycyrrhiza inflata [J]. Bioorg Med Chem. 1998, 6(3): 339-347.
[25]吴碧华,龙存国,王晓明,等.甘草总黄酮清除羟自由基作用的体外实验探讨[J].川北医学院学报. 2001, 16(1): 3-5.
[26]傅乃武,刘朝阳,张如意,等.甘草黄酮类和三萜类化合物抗氧化作用的研究[J].中药药理与临床. 1994, 10(5): 26-29.
[27] Tamir, S, Eizenberg, M, Somjen, D, et al. Estrogen-like activity of glabrene and other constituents isolated from licorice root [J]. J Steroid Biochem Mol Biol. 2001, 78(3): 291-298.
[28] Hoyoka, N. Nutraceuticals: Des FoodsⅢ: Garlic, Soy Licorice. [M]. 3rd. 1997: 279-285.
[29] Jackson, K M, Deleon, M. Dibenzoylmethane induces cell cycle deregulation in human prostate cancer cells [J]. Cancer Lett. 2002, 178: 161-165.
[30] Rafi, M M, Rosen, R T, A, V, et al. Modulation of bcl-2 and cytotoxicity by licochalcone-A, a novel estrogenic flavonoid [J]. Anticancer Res. 2000, 20: 2653-2658.
[31] Rafi, M M, Vastano, Z, Zhu, N, et al. Novel polyphenol molecule isolated from licorice root (Glycyrrhiza glabra) induces apoptosis, G2/M cell cycle arrest, and Bcl-2 phosphorylation in tumor cell lines [J]. Journal Agric Food Chem. 2002, 50: 677-684.
[32] Fu, Y, Hsieh, T C, Guo, J Q, et al. Licochalcone-A, a novel flavonoid isolated from licorice root (Glycyrrhiza glabra), causes G2 and late-G1 arrests in androgen-independent PC-3 prostate cancer cells [J]. Biochem Bioph Res Co. 2004, 322: 263-270.
[33] Fukai, T, Sakagami, H, Toguchi, M, et al. Cytotoxic activity of low molecular weight polyphenols against human oral tumor cell lines [J]. Anticancer Res. 2000, 20(4): 2525-2536.
[34]王秀梅,贾世山.甘草叶总黄酮和干扰素诱导单核巨噬细胞产生肿瘤坏死因子[J].中国中西医结合杂志. 1993, 13: 134.
[35]董菁,王秀梅.某草叶中黄酮成分体外诱生细胞毒因子的实验研究[J].汕头大学医学院学报. 1994(2): 9-11.
[36]黄华艺,查锡良.黄酮类化合物抗肿瘤作用的研究进展[J].中国新药与临床杂志. 2002, 21(7): 428-433.
[37]李铁民,梁再赋.甘草提取物及其衍生物的抗病毒研究现状[J].中草药. 1994, 25(12): 655-658.
[38]奥田拓男.日本药学会第108回药理会1989.
[39] Hatano, T, Kagawa, H, Yasuhara, T. Two new flavonoids and other constituents in licorice root their relative astringency and radical scavenging effects [J]. Chem Pharm Bull. 1988, 36(6): 2090-2098.
[40]陈蕙芳.植物活性成分大辞典[M].第三册.北京:中国医药科技出版社, 2001: 711-713.
[41]来国防,陈纪军,王易芬.黄酮类化合物抗HIV活性研究[J].商丘师范学院学报. 2002, 18(5): 83-87.
[42]王根生,韩哲武.甘草类黄酮对乙醇所致肝胀损伤的影响[J].中国药理学通报. 1993, 9(4): 271-273.
[43]贾国惠,贾世山.甘草中黄酮的药理作用研究进展[J].中国药学杂志. 1998, 33(9): 513-516.
[44] Fukai, T, Marumo, A, Kaitou, K, et al. Anti-Helicobacter pylori flavonoids from licorice extract [J]. Life Science. 2002, 71(12): 1449-1463.
[45] Tsukiyama, R, Katsura, H, Tokuriki, N, et al. Antibacterial activity of licochalcone A against spore-forming bacteria [J]. Antimicrob Agents Ch. 2002, 46(5): 1226-1230.
[46]胡小鹰,彭国平.甘草总黄酮抗心律失常作用研究[J].中草药. 1996, 27(12): 733-736.
[47]谢世荣,黄彩云,杨静娴.甘草黄酮抗实验性心律失常的作用[J].基础医学与临床. 1998, 18(2): 72-74.
[48] Shibata, S, Inoue, H, Iwata, S, et al. Inhibitory effects of licochalcone A isolatedfrom Glycyrrhiza inflata root on inflammatory ear edema and tumour promotion in mice, Planta Med [J]. Planta Med. 1991, 57(3): 221-224.
[49] Kimura, Y, Okuda, H, Okuda, T. Effects of chalcones isolated from licorice roots on leukotriene biosynthesis in human polymorphonuclear neutrophils [J]. Phytother Res. 1988, 2(3): 140-145.
[50] Yokota, T, Nishio, H, Kubota, Y, et al. The inhibitory effect of glabridin from licorice extracts on melanogenesisand inflammation [J]. Pigment Cell Res. 1998, 11(6): 335-361.
[51] Tawata, M, Aida, K. Anti-platelet action of isoliquiritigenin, an aldose redutase inhibitor in licorice [J]. Eur Journal Pharmacol. 1992, 212: 87-92.
[52] Tawata, M, Yoda, Y. Anti-platelet section of Gu-7, a 3-arylcoumarin derivative, purified from Glycryrrhiza radix [J]. Planta Med. 1990, 56(3): 259-266.
[53] Somjen, D, Katzburg, S, Vaya, J, et al. Estrogenic activity of glabridin and glabrene from licorice roots on human osteoblasts and prepubertal rat skeletal tissues [J]. Journal of Steroid Biochemistry & Molecular Biology. 2004, 91: 241-246.
[54]徐良,步平.美白祛斑化妆品及其未来发展[J].日用化学工业. 2001, 31(2): 42-45.
[55]吴志刚,吴书.美白新纪元[J].中国化妆品. 2001, 92(20): 20-28.
[56]李洪林,沈建华.虚拟筛选与新药发现[J].生命科学. 2005, 12(2): 125-131.
[57] Rarey, D M, Kramer, B, Lengauer, T, et al. A fast flexible docking method using an incremental construction algorithm [J]. Journal of Molecule Biology. 1996, 261: 470-489.
[58] Morris, G M, Goodsell, D S, Halliday, R S, et al. Automated docking using a lamarckian genetic algorithm and empirical binding free energy function [J]. Journal of Computer Chemistry. 1998, 19(1): 1639-1662.
[59] Jones, G, Willett, P, Glen, R C, et al. Development and validation of a genetic algorithm for flexible docking [J]. Journal of Molecule Biology. 1997, 267(7): 400-403.
[60] Li, H, Li, C, Gui, C, et al. GAsDock: a new approach for rapid flexible dockingbased on an improved multi-population genetic algorithm [J]. Bioorg Med Chem Lett. 2004, 14(2): 170-176.
[61] Kuntz, I D, Blaney, J M, Oatley, S J, et al. A geometric approach to macromolecule-ligand interactions [J]. Journal of Molecule Biology. 1982, 161(269-288): 629-637.
[62] Silvia, S, Efrem, C, Gianluca, D P. Polymeric hydrophobic membranes as a tool to control polymorphism and protein–ligand interactions [J]. J Membrane Sci. 2006, 131(2): 1149-1153.
[63]罗国安,王义明,饶毅.中药中成药现代化研究[J].中成药. 2000, 22(1): 71-79.
[64] Lazarowych, N. Use of fingerprinting and maker compounds for identification and standardization of botanical [J]. Drug Inf J. 1998, 32: 497-512.
[65] Bauer, R. Quality criteria and phytopharmaceuticals: can acceptable drug standards be achieved [J]. Drug Inf J. 1998, 32: 101-110.
[66] Barl, B. Quality analysis and standardized extracts of medicinal herbs[C]. University of Saskatchewan: 1997.
[67]谢培山.中药色谱指纹图谱北京:人民卫生出版社, 2005: 13-15.
[68] Haraguchi, H, Yoshida, N, Ishikawa, H, et al. Protection of mitochondrial functions against oxidative stresses by isoflavans from Glycyrrhiza glabra [J]. J Pharm Pharmacol. 2000, 52(2): 219-223.
[69] Belinky, P A, Aviram, M, Mahmood, S, et al. Structural aspects of the inhibitory effect of glabridin on LDL oxidation [J]. Free Radic Biol Med. 1998, 24(9): 1419-1429.
[70] Belinky, P A, Aviram, M, Fuhrman, B, et al. The antioxidative effects of the isoflavan glabridin on endogenous constituents of LDL during its oxidation [J]. Atherosclerosis. 1998, 137(1): 49-61.
[71] Fuhrman, B, Buch, S, Vaya, J, et al. Licorice extract and its major polyphenol glabridin protect low-density lipoprotein against lipid peroxidation: in vitro and ex vivo studies in humans and in atherosclerotic apolipoprotein E-deficient mice [J]. Am J Clin Nutr. 1997, 66: 267-275.
[72] Shibata, S. Anti-tumorigenic chalcones [J]. Stem Cells. 1994, 12(1): 44-52.
[73] Iwata, S, Nishino, T, Nagata, N, et al. Antitumorigenic activities of chalcones. I: Inhibitory effects of chalcone derivatives on 32Pi-incorporation into phospholipids of HeLa cells promoted by 12-O-tetradecanoyl-phorbol 13-acetate (TPA) [J]. Biol Pharm Bull. 1995, 18(12): 1710-1713.
[74] De Vincenzo, R, Scambia, G, Benedetti Panici, P, et al. Effect of synthetic and naturally occurring chalcones on ovarian cancer cell growth: structure-activity relationships [J]. Anticancer Drug Des. 1995, 10(6): 481-490.
[75] Iwata, S, Nishino, T, Inoue, H, et al. Antitumorigenic activities of chalcones (II). Photo-isomerization of chalcones and the correlation with their biological activities [J]. Biol Pharm Bull. 1997, 20(12): 1266-1270.
[76] Fukai, T, Marumo, A, Kaitou, K, et al. Anti-Helicobacter pylori flavonoids from licorice extract [J]. Life Sci. 2002, 71(12): 1449-1463.
[77] Tamir, S, Eizenberg, M, Somjen, D, et al. Estrogen-like activity of glabrene and other constituents isolated from licorice root [J]. J Steroid Biochem Mol Biol. 2001, 78(3): 291-298.
[78] Somjen, D, Knoll, E, Vaya, J, et al. Estrogen-like activity of licorice root constituents: glabridin and glabrene, in vascular tissues in vitro and in vivo [J]. J Steroid Biochem Mol Biol. 2004, 91(3): 147-155.
[79]韩博,陈文,翟科峰,等.甘草苷作对照品测定乌拉尔甘草中总黄酮的含量[J].石河子大学学报. 2006, 24(4): 472-474.
[80]闫永红,王文全,杨娜.栽培甘草中总黄酮的含量测定[J].中国中药杂志. 2006, 31(12): 1019-1021.
[81] Vaya, J, Belinky, P A, Aviram, M. Antioxidant constituents from licorice roots: isolation, structure elucidation and antioxidative capacity toward LDL oxidation [J]. Free Rad Biol Med. 1997, 23(2): 302-313.
[82] Kanazawa, M, Satomi, Y, Mizutani, Y. Isoliquiritigeni inhibits the growth of prostate cancer [J]. Eur Urol. 2003, 43(5): 580-586.
[83] Tamira, S, Eizenberg, M. Estrogen-like activity of glabrene and other constituents isolated from licorice root [J]. Journal of Steroid Biochemistry &Molecular Biology. 2001, 78: 291-295.
[84] Toshio, F, Ai, M, Kiyoshi, K. Antimicrobial activity of licorice flavonoids against methicillin-resistant Staphylococcus aureus [J]. Fitoterapia. 2002, 73(6): 536-539.
[85] Fuhrman, B, Volkova, N, Kaplan, M, et al. Antiatherosclerotic effects of licorice extracts supplementation on hypercholesterolemic patients: increased resistance of LDL to atherogenic modifications, reduced plasma lipid levels, and decreased systolic blood pressure [J]. Nutrition. 2002, 18: 268-273.
[86]付玉杰,施晓光,刘晓娜,等.超临界CO2提取甘草地上部分总黄酮[J].植物研究. 2007, 27(3): 372-375.
[87]赵兵.中药提取分离新技术[J].北方牧业. 2007, 6: 8-8.
[88]郭春晓,王世宽.高新技术在天然产物有效成分分离中的应用[J].化学与生物工程. 2006, 23(5): 38-40.
[89]何春霞,苏力坦阿巴白克力. AB-8大孔树脂纯化欧洲鳞毛蕨总黄酮的研究[J].生物技术. 2007, 17(2): 57-59.
[90]曹群华,瞿伟菁,李家贵,等.大孔树脂吸附纯化沙棘籽渣总黄酮的研究[J].中国中药杂志. 2004, 29(3): 225-228.
[91] Ghelardini, C, Galeotti, N, Barboloni, A, et al. Memory facilitation and stimulation of endogenous nerve growth factor synthesis by the acetylcholine releaser PG-9 [J]. J Pharmacol. 1998, 78(3): 245-251.
[92] Bhattacharya, S K, Upadhyay, S N, Jaiswal, A K. Effect of piracetam on electroshock induced amnesia and decrease in brain acetylcholine in rats [J]. Indian J Exp Biol. 1993, 31(1): 822-824.
[93] Dhingra, D, Parle, M, Kulkarni, S K. Comparative brain cholinesterase inhibiting activity of Glycyrrhiza glabra, Myristica fragrans, ascorbic acid and metrifonate in mice [J]. J Med Food. 2006, 9(2): 281-283.
[94] Vasudevan, M, Parle, M. Pharmacological actions of Thespesia populnea relevant to Alzheimer's disease [J]. Phytomedicine. 2006, 13(9): 677-687.
[95] Parle, M, Dhingra, D, Kulkarni, S K. Memory-strengthening activity ofGlycyrrhiza glabra in exteroceptive and interoceptive behavioral models [J]. J Med Food. 2004, 7(4): 462-466.
[96] Dhingra, D, Parle, M, Kulkarni, S K. Memory enhancing activity of Glycyrrhiza glabra in mice [J]. J Ethnopharmacol. 2004, 91(2): 361-365.
[97] Ravelli, R B, Raves, M L, Ren, Z, et al. Static Laue diffraction studies on acetylcholinesterase [J]. Acta Crystallogr Sect D. 1998, 54: 1359-1366.
[98] Krkinjuntti, T, Kurz, A, Gauthier, S. Efficacy of galantamine in probable vascular dementia and Alzheimer's disease combined with cerebrovascular disease: A randomized trial [J]. Lancet. 2002, 359: 1283-1291.
[99] Rosenblat, M, Belinky, P, Vaya, J, et al. Macrophage enrichment with the isoflavan glabridin inhibits NADPH oxidase-induced cell-mediated oxidation of low density lipoprotein: A possible role for kinase C. [J]. J Biol Chem. 1999, 274(20): 13790-13799.
[100] Fukai, T, Satoh, K, Nomura, T, et al. Antinephritis and radical scavenging activity of prenylflavonoids [J]. Fitoterapia. 2003, 74(7): 720-724.
[101] Tamir, S, Eizenberg, M, Somjen, D, et al. Estrogenic and antiproliferative properties of glabridin from licorice in human breast cancer cells [J]. Cancer Res. 2000, 60(20): 5704-5709.
[102] Nerya, O, Vaya, J, Musa, R, et al. Glabrene and isoliquiritigenin as tyrosinase inhibitors from licorice roots [J]. J Agric Food Chem. 2003, 51(5): 1201-1207.
[103] Rauchensteiner, F, Matsumura, Y, Yamamoto, Y, et al. Analysis and comparision of radix Glycyrrhizae (licorice) from europe and china by capillary-zone electrophoresis (CZE) [J]. J Pharmaceut Biomed. 2005, 38: 594-600.
[104] Rao, S K, Andrade, C, Reddy, K. Memory protective effect of indomethacin against electroconvulsive shock-induced retrograde amnesia in rats [J]. Biol Psychiat. 2002, 51(9): 770-773.
[105] Shibata, S. A drug over the millennia: pharmacognosy, chemistry, and pharmacology of licorice [J]. Yakugaku Zasshi. 2000, 120(10): 849-862.
[106] Kajiyama, K, Demizu, S, Hiraga, Y, et al. Two prenylated retrochalcones fromGlycyrrhiza inflata [J]. Phytochemistry. 1992, 31(9): 3229-3232.
[107] Haraguchi, H, Tanimoto, K, Tamura, Y, et al. Mode of antibacterial action of retrochalcones from Glycyrrhiza inflata [J]. Phytochemistry. 1998, 48(1): 125-129.
[108] Okada, K, Tamura, Y, Yamamoto, M, et al. Identification of antimicrobial and antioxidant constituents from licorice of Russian and Xinjiang origin [J]. Chem Pharm Bull. 1989, 37(9): 2528-2530.
[109] Chen, M, Zhai, L, Christensen, S B, et al. Inhibition of fumarate reductase in Leishmania major and L. donovani by chalcones [J]. Antimicrob Agents Chemother. 2001, 45(7): 2023-2029.
[110] Chen, M, Christensen, S B, Blom, J, et al. Licochalcone A, a novel antiparasitic agent with potent activity against human pathogenic protozoan species of Leishmania [J]. Agents Chemother. 1993, 37(12): 2550-2556.
[111] Zhai, L, Blom, J, Chen, M, et al. The antileishmanial agent licochalcone A interferes with the function of parasite mitochondria [J]. Antimicrob Agents Chemother. 1995, 39(12): 2742-2748.
[112] Zhai, L, Chen, M, Blom, J, et al. The antileishmanial activity of novel oxygenated chalcones and their mechanism of action [J]. J Antimicrob Chemother. 1999, 43(6): 793-803.
[113] Rafi, M M, Rosen, R T, Vassil, A, et al. Modulation of bcl-2 and cytotoxicity by licochalcone-A, a novel estrogenic flavonoid [J]. Anticancer Res. 2000, 20(4): 2653-2658.
[114] Wang, Q E, Lee, S C, Wang, X R. Isolation and purification of inflacoumarin A and licochalcone A from licorice by high-speed counter-current chromatography [J]. J Chromatogr A. 2004, 1048(1): 51-57.
[115] Atta, A H, Alkohafi, A. Antinociceptive and anti-inflammatory effects of some Jordanian medicinal plants extracts [J]. J Ethnopharmacol. 1998, 60(2): 117-124.
[116] Shu, X S, Gao, Z H, Yang, X L. Anti-inflammatory and anti-nociceptive activities of Smilax china L. aqueous extract [J]. J Ethnopharmacol. 2006, 103(3): 327-332.
[117] Portanova, P J, Zhang, Y, Anderson, G D. Selective neutralization of prostaglandin E2 blocks inflammation, hyperalgesia and interleukin 6 production in vivo [J]. J Exp Med. 1996, 184(3): 833-891.
[118] Subbaramaiah K., Telang N., Ramonetti, J T, Araki, R, Devito, B, et al. Transcription of cyclooxygenase-2 is enhanced in transformed mammary epithelial cells [J]. Cancer Res. 1996, 56: 4424-4429.
[119] Lin, C N, Lee, T H, Hsu, M F, et al. 2',5'-Dihydroxychalcone as a potent chemical mediator and cyclooxygenase inhibitor [J]. J Pharm Pharmacol. 1997, 49(5): 530-536.
[120]李晓波,屠鹏飞.中药指纹图谱体系[J].中草药. 2003, 34(5): 385-388.
[121] Fda. Guidence for industry: Botanical Drug Products (Draft Guidence)[C]. 2000.
[122] Who. Guidence for the Assessment of Herbal Medicines[C]. 1996.
[123]王月辉,乔斌,蒋建兰,等.甘草药材HPLC指纹图谱的研究[J].化学工业与工程. 2006, 23(3): 250-253.
[124]吴昭晖,罗佳波,游文玮.甘草药材HPLC指纹图谱研究[J].中草药. 2005, 36(12): 1868-1872.
[125]彭励,胡正海.甘草生物学及化学成分的研究进展[J].中草药. 2005, 36(11): 1744-1747.
[126]屠鹏飞.高效液相法制定中药材和中药注射剂特征指纹图谱的探讨[J].中成药. 2000, 22(7): 516-519.
[2]郑虎占,董泽宏,余靖.中药现代研究与应用[M].北京:学苑出版社, 1997, 1256-1279.
[3]周成明. 80种常用中草药栽培[M].第二版;北京:中国农业出版社, 2002, 1-4.
[4] Ichimura, M, Furuno, T, Takahashi, T, et al. Enzymic O-methylation of isoliquiritigenin and licodione in alfalfa and licorice culture [J]. Phytochemistry. 1997, 44(6): 991-995.
[5]杜等.甘草毛状根培养系统的建立及化学成分分析[J].植物资源与环境学报. 2001, 10(1): 7-10.
[6]郭江平.乌拉尔甘草大面积人工种植试验初报[J].干旱地区农业研究. 2000, 18(4): 133-136.
[7]于林清,何茂泰,王照兰.甘草组织培养快速繁殖技术研究[J].中国草地. 1999, 18(1): 12-14.
[8]殷生章.一个新的二氢黄酮苷[J].中国药科大学学报. 1999, 30(1): 19-20.
[9] Hatano, T, Shintani, Y, Aga, Y. Phenolics constituents of licoriceⅧ. Structures of glicophenone and glicoisoflavanone and effects of licorice phenolics on methicillin-resistant stanphlococcus aureus [J]. Chem Pharm Bull. 2000, 48(9): 1286-1292.
[10] Hatano, T, Takagi, M, Ito, H. Phenolic constituents of liquoriceⅦ. A new chalcone with a potent radical activity and accompanying phenolics from liquorice [J]. Chem Pharm Bull. 1997, 45(9): 1485-1492.
[11] Shibano, M, Matsumoto, Y, Nakao, E. Studies on the index compounds for HPLC analysis of Glycyrrhiza uralensis [J]. Heterocycles. 1997, 45(10): 2053-2060.
[12] Kinoshita, T, Kajiyama, K, Hiraga, Y. Isoflavan derivaties from Glycyrrhizaglabra [J]. Heterocycles. 1996, 43(3): 581-587.
[13] Fukai, T, Tantai, L, Nomura, T. Isoprenoid-substituted flavonoids from Glycyrrhiza glabra [J]. Heterocycles. 1996, 43(2): 531-532.
[14] Fukai, T, Cai, B, Maruno, K. "Phenolic costituents of Glycyrrhiza species" An isoprenylated flavanone from Glycyrrhiza glabra and recassay of licorice phenols [J]. Phytochemistry. 1998, 49(7): 2005-2013.
[15] Asada, Y, Li, W, Yoshikawa. The first prenylated biaurone, licoagrone from hairy root cultures of Glycyrrhiza glabra [J]. Phytochemistry. 1999, 50(6): 1015-1019.
[16] Li, W, Asada, Y, Yoshikawa, T. Antimicrobial flavonoids from Glycyrrhiza glabra hairy root culture [J]. Planta Med. 1998, 64(8): 746-747.
[17]王邠,邹坤.胀果甘草中2个新的二氢黄酮甙[J].药学学报. 1997, 32(3): 199-202.
[18] Fukai, T, Nomura, T. Isoprenoid-substituted flavonoids from roots of Glycyrrhiza inflata [J]. Phytochemistry. 1995, 38(3): 759-765.
[19]李伟东,阚毓铭.刺果甘草化学成分的研究[J].中国药学杂志. 1999, 34(1): 11-14.
[20]张聿梅,许旭东,胡碧煌.黄甘草异黄酮成分的研究[J].药学学报. 1997, 32(4): 301-304.
[21] Kitagawa, I, Chen, W, Hori, K. Chemical studies of chinese licorice-rootsⅡ. Five new flavonoid constituents from the roots of Glycyrrhiza aspera pall collected in xinjiang [J]. Chem Pharm Bull. 1998, 46(10): 1511-1517.
[22] Fuhrman, B, Volkova, N. Antiatherosclerotic effects of licorice extracts supplementation on hypercholesterolemic patients: increased resistance of LDL to atherogenic modifications, reduced plasma plasma lipid levels, and decreased systolic blood pressure. [J]. Nutrition. 2002, 18: 268-273.
[23] Vaya, J, Belinky, P A, Aviram, M. Antioxidant constituents from licorice roots: isolation, structure elucidation and antioxidative capacity toward LDL oxidation [J]. Free Rad Biol Med. 1997, 23(2): 302-313.
[24] HaraguchiH;ishikawaH, MizutaniK;TamuraY;kinoshitaT. Antioxidative andsuperoxide scavenging activities of retrochalcones in Glycyrrhiza inflata [J]. Bioorg Med Chem. 1998, 6(3): 339-347.
[25]吴碧华,龙存国,王晓明,等.甘草总黄酮清除羟自由基作用的体外实验探讨[J].川北医学院学报. 2001, 16(1): 3-5.
[26]傅乃武,刘朝阳,张如意,等.甘草黄酮类和三萜类化合物抗氧化作用的研究[J].中药药理与临床. 1994, 10(5): 26-29.
[27] Tamir, S, Eizenberg, M, Somjen, D, et al. Estrogen-like activity of glabrene and other constituents isolated from licorice root [J]. J Steroid Biochem Mol Biol. 2001, 78(3): 291-298.
[28] Hoyoka, N. Nutraceuticals: Des FoodsⅢ: Garlic, Soy Licorice. [M]. 3rd. 1997: 279-285.
[29] Jackson, K M, Deleon, M. Dibenzoylmethane induces cell cycle deregulation in human prostate cancer cells [J]. Cancer Lett. 2002, 178: 161-165.
[30] Rafi, M M, Rosen, R T, A, V, et al. Modulation of bcl-2 and cytotoxicity by licochalcone-A, a novel estrogenic flavonoid [J]. Anticancer Res. 2000, 20: 2653-2658.
[31] Rafi, M M, Vastano, Z, Zhu, N, et al. Novel polyphenol molecule isolated from licorice root (Glycyrrhiza glabra) induces apoptosis, G2/M cell cycle arrest, and Bcl-2 phosphorylation in tumor cell lines [J]. Journal Agric Food Chem. 2002, 50: 677-684.
[32] Fu, Y, Hsieh, T C, Guo, J Q, et al. Licochalcone-A, a novel flavonoid isolated from licorice root (Glycyrrhiza glabra), causes G2 and late-G1 arrests in androgen-independent PC-3 prostate cancer cells [J]. Biochem Bioph Res Co. 2004, 322: 263-270.
[33] Fukai, T, Sakagami, H, Toguchi, M, et al. Cytotoxic activity of low molecular weight polyphenols against human oral tumor cell lines [J]. Anticancer Res. 2000, 20(4): 2525-2536.
[34]王秀梅,贾世山.甘草叶总黄酮和干扰素诱导单核巨噬细胞产生肿瘤坏死因子[J].中国中西医结合杂志. 1993, 13: 134.
[35]董菁,王秀梅.某草叶中黄酮成分体外诱生细胞毒因子的实验研究[J].汕头大学医学院学报. 1994(2): 9-11.
[36]黄华艺,查锡良.黄酮类化合物抗肿瘤作用的研究进展[J].中国新药与临床杂志. 2002, 21(7): 428-433.
[37]李铁民,梁再赋.甘草提取物及其衍生物的抗病毒研究现状[J].中草药. 1994, 25(12): 655-658.
[38]奥田拓男.日本药学会第108回药理会1989.
[39] Hatano, T, Kagawa, H, Yasuhara, T. Two new flavonoids and other constituents in licorice root their relative astringency and radical scavenging effects [J]. Chem Pharm Bull. 1988, 36(6): 2090-2098.
[40]陈蕙芳.植物活性成分大辞典[M].第三册.北京:中国医药科技出版社, 2001: 711-713.
[41]来国防,陈纪军,王易芬.黄酮类化合物抗HIV活性研究[J].商丘师范学院学报. 2002, 18(5): 83-87.
[42]王根生,韩哲武.甘草类黄酮对乙醇所致肝胀损伤的影响[J].中国药理学通报. 1993, 9(4): 271-273.
[43]贾国惠,贾世山.甘草中黄酮的药理作用研究进展[J].中国药学杂志. 1998, 33(9): 513-516.
[44] Fukai, T, Marumo, A, Kaitou, K, et al. Anti-Helicobacter pylori flavonoids from licorice extract [J]. Life Science. 2002, 71(12): 1449-1463.
[45] Tsukiyama, R, Katsura, H, Tokuriki, N, et al. Antibacterial activity of licochalcone A against spore-forming bacteria [J]. Antimicrob Agents Ch. 2002, 46(5): 1226-1230.
[46]胡小鹰,彭国平.甘草总黄酮抗心律失常作用研究[J].中草药. 1996, 27(12): 733-736.
[47]谢世荣,黄彩云,杨静娴.甘草黄酮抗实验性心律失常的作用[J].基础医学与临床. 1998, 18(2): 72-74.
[48] Shibata, S, Inoue, H, Iwata, S, et al. Inhibitory effects of licochalcone A isolatedfrom Glycyrrhiza inflata root on inflammatory ear edema and tumour promotion in mice, Planta Med [J]. Planta Med. 1991, 57(3): 221-224.
[49] Kimura, Y, Okuda, H, Okuda, T. Effects of chalcones isolated from licorice roots on leukotriene biosynthesis in human polymorphonuclear neutrophils [J]. Phytother Res. 1988, 2(3): 140-145.
[50] Yokota, T, Nishio, H, Kubota, Y, et al. The inhibitory effect of glabridin from licorice extracts on melanogenesisand inflammation [J]. Pigment Cell Res. 1998, 11(6): 335-361.
[51] Tawata, M, Aida, K. Anti-platelet action of isoliquiritigenin, an aldose redutase inhibitor in licorice [J]. Eur Journal Pharmacol. 1992, 212: 87-92.
[52] Tawata, M, Yoda, Y. Anti-platelet section of Gu-7, a 3-arylcoumarin derivative, purified from Glycryrrhiza radix [J]. Planta Med. 1990, 56(3): 259-266.
[53] Somjen, D, Katzburg, S, Vaya, J, et al. Estrogenic activity of glabridin and glabrene from licorice roots on human osteoblasts and prepubertal rat skeletal tissues [J]. Journal of Steroid Biochemistry & Molecular Biology. 2004, 91: 241-246.
[54]徐良,步平.美白祛斑化妆品及其未来发展[J].日用化学工业. 2001, 31(2): 42-45.
[55]吴志刚,吴书.美白新纪元[J].中国化妆品. 2001, 92(20): 20-28.
[56]李洪林,沈建华.虚拟筛选与新药发现[J].生命科学. 2005, 12(2): 125-131.
[57] Rarey, D M, Kramer, B, Lengauer, T, et al. A fast flexible docking method using an incremental construction algorithm [J]. Journal of Molecule Biology. 1996, 261: 470-489.
[58] Morris, G M, Goodsell, D S, Halliday, R S, et al. Automated docking using a lamarckian genetic algorithm and empirical binding free energy function [J]. Journal of Computer Chemistry. 1998, 19(1): 1639-1662.
[59] Jones, G, Willett, P, Glen, R C, et al. Development and validation of a genetic algorithm for flexible docking [J]. Journal of Molecule Biology. 1997, 267(7): 400-403.
[60] Li, H, Li, C, Gui, C, et al. GAsDock: a new approach for rapid flexible dockingbased on an improved multi-population genetic algorithm [J]. Bioorg Med Chem Lett. 2004, 14(2): 170-176.
[61] Kuntz, I D, Blaney, J M, Oatley, S J, et al. A geometric approach to macromolecule-ligand interactions [J]. Journal of Molecule Biology. 1982, 161(269-288): 629-637.
[62] Silvia, S, Efrem, C, Gianluca, D P. Polymeric hydrophobic membranes as a tool to control polymorphism and protein–ligand interactions [J]. J Membrane Sci. 2006, 131(2): 1149-1153.
[63]罗国安,王义明,饶毅.中药中成药现代化研究[J].中成药. 2000, 22(1): 71-79.
[64] Lazarowych, N. Use of fingerprinting and maker compounds for identification and standardization of botanical [J]. Drug Inf J. 1998, 32: 497-512.
[65] Bauer, R. Quality criteria and phytopharmaceuticals: can acceptable drug standards be achieved [J]. Drug Inf J. 1998, 32: 101-110.
[66] Barl, B. Quality analysis and standardized extracts of medicinal herbs[C]. University of Saskatchewan: 1997.
[67]谢培山.中药色谱指纹图谱北京:人民卫生出版社, 2005: 13-15.
[68] Haraguchi, H, Yoshida, N, Ishikawa, H, et al. Protection of mitochondrial functions against oxidative stresses by isoflavans from Glycyrrhiza glabra [J]. J Pharm Pharmacol. 2000, 52(2): 219-223.
[69] Belinky, P A, Aviram, M, Mahmood, S, et al. Structural aspects of the inhibitory effect of glabridin on LDL oxidation [J]. Free Radic Biol Med. 1998, 24(9): 1419-1429.
[70] Belinky, P A, Aviram, M, Fuhrman, B, et al. The antioxidative effects of the isoflavan glabridin on endogenous constituents of LDL during its oxidation [J]. Atherosclerosis. 1998, 137(1): 49-61.
[71] Fuhrman, B, Buch, S, Vaya, J, et al. Licorice extract and its major polyphenol glabridin protect low-density lipoprotein against lipid peroxidation: in vitro and ex vivo studies in humans and in atherosclerotic apolipoprotein E-deficient mice [J]. Am J Clin Nutr. 1997, 66: 267-275.
[72] Shibata, S. Anti-tumorigenic chalcones [J]. Stem Cells. 1994, 12(1): 44-52.
[73] Iwata, S, Nishino, T, Nagata, N, et al. Antitumorigenic activities of chalcones. I: Inhibitory effects of chalcone derivatives on 32Pi-incorporation into phospholipids of HeLa cells promoted by 12-O-tetradecanoyl-phorbol 13-acetate (TPA) [J]. Biol Pharm Bull. 1995, 18(12): 1710-1713.
[74] De Vincenzo, R, Scambia, G, Benedetti Panici, P, et al. Effect of synthetic and naturally occurring chalcones on ovarian cancer cell growth: structure-activity relationships [J]. Anticancer Drug Des. 1995, 10(6): 481-490.
[75] Iwata, S, Nishino, T, Inoue, H, et al. Antitumorigenic activities of chalcones (II). Photo-isomerization of chalcones and the correlation with their biological activities [J]. Biol Pharm Bull. 1997, 20(12): 1266-1270.
[76] Fukai, T, Marumo, A, Kaitou, K, et al. Anti-Helicobacter pylori flavonoids from licorice extract [J]. Life Sci. 2002, 71(12): 1449-1463.
[77] Tamir, S, Eizenberg, M, Somjen, D, et al. Estrogen-like activity of glabrene and other constituents isolated from licorice root [J]. J Steroid Biochem Mol Biol. 2001, 78(3): 291-298.
[78] Somjen, D, Knoll, E, Vaya, J, et al. Estrogen-like activity of licorice root constituents: glabridin and glabrene, in vascular tissues in vitro and in vivo [J]. J Steroid Biochem Mol Biol. 2004, 91(3): 147-155.
[79]韩博,陈文,翟科峰,等.甘草苷作对照品测定乌拉尔甘草中总黄酮的含量[J].石河子大学学报. 2006, 24(4): 472-474.
[80]闫永红,王文全,杨娜.栽培甘草中总黄酮的含量测定[J].中国中药杂志. 2006, 31(12): 1019-1021.
[81] Vaya, J, Belinky, P A, Aviram, M. Antioxidant constituents from licorice roots: isolation, structure elucidation and antioxidative capacity toward LDL oxidation [J]. Free Rad Biol Med. 1997, 23(2): 302-313.
[82] Kanazawa, M, Satomi, Y, Mizutani, Y. Isoliquiritigeni inhibits the growth of prostate cancer [J]. Eur Urol. 2003, 43(5): 580-586.
[83] Tamira, S, Eizenberg, M. Estrogen-like activity of glabrene and other constituents isolated from licorice root [J]. Journal of Steroid Biochemistry &Molecular Biology. 2001, 78: 291-295.
[84] Toshio, F, Ai, M, Kiyoshi, K. Antimicrobial activity of licorice flavonoids against methicillin-resistant Staphylococcus aureus [J]. Fitoterapia. 2002, 73(6): 536-539.
[85] Fuhrman, B, Volkova, N, Kaplan, M, et al. Antiatherosclerotic effects of licorice extracts supplementation on hypercholesterolemic patients: increased resistance of LDL to atherogenic modifications, reduced plasma lipid levels, and decreased systolic blood pressure [J]. Nutrition. 2002, 18: 268-273.
[86]付玉杰,施晓光,刘晓娜,等.超临界CO2提取甘草地上部分总黄酮[J].植物研究. 2007, 27(3): 372-375.
[87]赵兵.中药提取分离新技术[J].北方牧业. 2007, 6: 8-8.
[88]郭春晓,王世宽.高新技术在天然产物有效成分分离中的应用[J].化学与生物工程. 2006, 23(5): 38-40.
[89]何春霞,苏力坦阿巴白克力. AB-8大孔树脂纯化欧洲鳞毛蕨总黄酮的研究[J].生物技术. 2007, 17(2): 57-59.
[90]曹群华,瞿伟菁,李家贵,等.大孔树脂吸附纯化沙棘籽渣总黄酮的研究[J].中国中药杂志. 2004, 29(3): 225-228.
[91] Ghelardini, C, Galeotti, N, Barboloni, A, et al. Memory facilitation and stimulation of endogenous nerve growth factor synthesis by the acetylcholine releaser PG-9 [J]. J Pharmacol. 1998, 78(3): 245-251.
[92] Bhattacharya, S K, Upadhyay, S N, Jaiswal, A K. Effect of piracetam on electroshock induced amnesia and decrease in brain acetylcholine in rats [J]. Indian J Exp Biol. 1993, 31(1): 822-824.
[93] Dhingra, D, Parle, M, Kulkarni, S K. Comparative brain cholinesterase inhibiting activity of Glycyrrhiza glabra, Myristica fragrans, ascorbic acid and metrifonate in mice [J]. J Med Food. 2006, 9(2): 281-283.
[94] Vasudevan, M, Parle, M. Pharmacological actions of Thespesia populnea relevant to Alzheimer's disease [J]. Phytomedicine. 2006, 13(9): 677-687.
[95] Parle, M, Dhingra, D, Kulkarni, S K. Memory-strengthening activity ofGlycyrrhiza glabra in exteroceptive and interoceptive behavioral models [J]. J Med Food. 2004, 7(4): 462-466.
[96] Dhingra, D, Parle, M, Kulkarni, S K. Memory enhancing activity of Glycyrrhiza glabra in mice [J]. J Ethnopharmacol. 2004, 91(2): 361-365.
[97] Ravelli, R B, Raves, M L, Ren, Z, et al. Static Laue diffraction studies on acetylcholinesterase [J]. Acta Crystallogr Sect D. 1998, 54: 1359-1366.
[98] Krkinjuntti, T, Kurz, A, Gauthier, S. Efficacy of galantamine in probable vascular dementia and Alzheimer's disease combined with cerebrovascular disease: A randomized trial [J]. Lancet. 2002, 359: 1283-1291.
[99] Rosenblat, M, Belinky, P, Vaya, J, et al. Macrophage enrichment with the isoflavan glabridin inhibits NADPH oxidase-induced cell-mediated oxidation of low density lipoprotein: A possible role for kinase C. [J]. J Biol Chem. 1999, 274(20): 13790-13799.
[100] Fukai, T, Satoh, K, Nomura, T, et al. Antinephritis and radical scavenging activity of prenylflavonoids [J]. Fitoterapia. 2003, 74(7): 720-724.
[101] Tamir, S, Eizenberg, M, Somjen, D, et al. Estrogenic and antiproliferative properties of glabridin from licorice in human breast cancer cells [J]. Cancer Res. 2000, 60(20): 5704-5709.
[102] Nerya, O, Vaya, J, Musa, R, et al. Glabrene and isoliquiritigenin as tyrosinase inhibitors from licorice roots [J]. J Agric Food Chem. 2003, 51(5): 1201-1207.
[103] Rauchensteiner, F, Matsumura, Y, Yamamoto, Y, et al. Analysis and comparision of radix Glycyrrhizae (licorice) from europe and china by capillary-zone electrophoresis (CZE) [J]. J Pharmaceut Biomed. 2005, 38: 594-600.
[104] Rao, S K, Andrade, C, Reddy, K. Memory protective effect of indomethacin against electroconvulsive shock-induced retrograde amnesia in rats [J]. Biol Psychiat. 2002, 51(9): 770-773.
[105] Shibata, S. A drug over the millennia: pharmacognosy, chemistry, and pharmacology of licorice [J]. Yakugaku Zasshi. 2000, 120(10): 849-862.
[106] Kajiyama, K, Demizu, S, Hiraga, Y, et al. Two prenylated retrochalcones fromGlycyrrhiza inflata [J]. Phytochemistry. 1992, 31(9): 3229-3232.
[107] Haraguchi, H, Tanimoto, K, Tamura, Y, et al. Mode of antibacterial action of retrochalcones from Glycyrrhiza inflata [J]. Phytochemistry. 1998, 48(1): 125-129.
[108] Okada, K, Tamura, Y, Yamamoto, M, et al. Identification of antimicrobial and antioxidant constituents from licorice of Russian and Xinjiang origin [J]. Chem Pharm Bull. 1989, 37(9): 2528-2530.
[109] Chen, M, Zhai, L, Christensen, S B, et al. Inhibition of fumarate reductase in Leishmania major and L. donovani by chalcones [J]. Antimicrob Agents Chemother. 2001, 45(7): 2023-2029.
[110] Chen, M, Christensen, S B, Blom, J, et al. Licochalcone A, a novel antiparasitic agent with potent activity against human pathogenic protozoan species of Leishmania [J]. Agents Chemother. 1993, 37(12): 2550-2556.
[111] Zhai, L, Blom, J, Chen, M, et al. The antileishmanial agent licochalcone A interferes with the function of parasite mitochondria [J]. Antimicrob Agents Chemother. 1995, 39(12): 2742-2748.
[112] Zhai, L, Chen, M, Blom, J, et al. The antileishmanial activity of novel oxygenated chalcones and their mechanism of action [J]. J Antimicrob Chemother. 1999, 43(6): 793-803.
[113] Rafi, M M, Rosen, R T, Vassil, A, et al. Modulation of bcl-2 and cytotoxicity by licochalcone-A, a novel estrogenic flavonoid [J]. Anticancer Res. 2000, 20(4): 2653-2658.
[114] Wang, Q E, Lee, S C, Wang, X R. Isolation and purification of inflacoumarin A and licochalcone A from licorice by high-speed counter-current chromatography [J]. J Chromatogr A. 2004, 1048(1): 51-57.
[115] Atta, A H, Alkohafi, A. Antinociceptive and anti-inflammatory effects of some Jordanian medicinal plants extracts [J]. J Ethnopharmacol. 1998, 60(2): 117-124.
[116] Shu, X S, Gao, Z H, Yang, X L. Anti-inflammatory and anti-nociceptive activities of Smilax china L. aqueous extract [J]. J Ethnopharmacol. 2006, 103(3): 327-332.
[117] Portanova, P J, Zhang, Y, Anderson, G D. Selective neutralization of prostaglandin E2 blocks inflammation, hyperalgesia and interleukin 6 production in vivo [J]. J Exp Med. 1996, 184(3): 833-891.
[118] Subbaramaiah K., Telang N., Ramonetti, J T, Araki, R, Devito, B, et al. Transcription of cyclooxygenase-2 is enhanced in transformed mammary epithelial cells [J]. Cancer Res. 1996, 56: 4424-4429.
[119] Lin, C N, Lee, T H, Hsu, M F, et al. 2',5'-Dihydroxychalcone as a potent chemical mediator and cyclooxygenase inhibitor [J]. J Pharm Pharmacol. 1997, 49(5): 530-536.
[120]李晓波,屠鹏飞.中药指纹图谱体系[J].中草药. 2003, 34(5): 385-388.
[121] Fda. Guidence for industry: Botanical Drug Products (Draft Guidence)[C]. 2000.
[122] Who. Guidence for the Assessment of Herbal Medicines[C]. 1996.
[123]王月辉,乔斌,蒋建兰,等.甘草药材HPLC指纹图谱的研究[J].化学工业与工程. 2006, 23(3): 250-253.
[124]吴昭晖,罗佳波,游文玮.甘草药材HPLC指纹图谱研究[J].中草药. 2005, 36(12): 1868-1872.
[125]彭励,胡正海.甘草生物学及化学成分的研究进展[J].中草药. 2005, 36(11): 1744-1747.
[126]屠鹏飞.高效液相法制定中药材和中药注射剂特征指纹图谱的探讨[J].中成药. 2000, 22(7): 516-519.