虎杖生药学研究
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摘要
蓼科植物虎杖Polygonum cuspidatum Sieb. et Zucc.,主要为温带亚洲分布,在中国广布于秦岭、长江以南,达南岭和西南,东北达朝、韩、日,多在2000m以下的常绿林或中生混交林林下溪边,环境适应性极强。中药虎杖为P. cuspidatum的干燥根及根茎,有利湿退黄,清热解毒,散瘀止痛,止咳化痰之功效；虎杖叶在民间有悠久的药食两用历史,具有祛风湿,解热毒之功效。近年来虎杖药材需求量不断增长,各地不仅开采利用野生资源,同时也尝试进行规模化栽培；从而导致市场流通的虎杖因产地、来源的多样、生长年限及采收时间的不同而品质不一。为了深入地探讨中药虎杖的质量评价方法,并为合理的利用资源提供依据,本论文通过分子鉴定、分析和药理三个方面研究工作的有机结合,系统研究了中药虎杖的真伪、优劣和活性成分。同时,为了进一步完善药用植物虎杖的资源利用,本论文对虎杖叶的成分进行了鉴定及对主要成分进行了含量测定,同时筛选了虎杖叶中的抗氧化成分并对抗氧化能力进行了测定,为药食两用的虎杖叶的开发利用提供科学指导。
对虎杖根的提取物及主要单体化合物进行了降糖降脂活性的体外实验。结果表明虎杖根中促进细胞葡萄糖摄取的成分主要为乙酸乙酯萃取物中的蒽醌类成分。羟基蒽醌类化合物大黄素、大黄素甲醚、6-羟基芦荟大黄素、芦荟大黄素、大黄酚、大黄酸为主要的活性成分；而羟基蒽醌的糖苷类化合物并无明显改善活性,因而初步推断蒽醌类化合物的游离羟基是促进细胞葡萄糖摄取的必要条件。虎杖根中抑制脂质堆积的活性成分主要集中在正丁醇部位,虎杖苷、金丝桃苷、大黄素-8-O-葡萄糖苷、大黄酸和芦荟大黄素为主要的活性成分。
本研究从分子水平应用叶绿体psbA-trnH基因对不同产地采集的24份虎杖样品(包括根14份、叶10份)和同科易混淆植物32条序列进行DNA的分子鉴定研究。对虎杖样品的种内变异情况做了统计,并采用了BLAST、最小距离法、NJ邻接法构建系统发育树来区分虎杖和同科易混淆植物。结果显示虎杖种内最小变异为0,最大变异为7个碱基变异及2个碱基的插入缺失,表明虎杖种内不同居群材料在分子水平上差异较小。而虎杖与同属及同科易混淆植物在分子水平上差异较大。证实psbA-trnH条形码序列能够准确鉴别虎杖药材的基原植物,同时也能准确区分虎杖药材与同科易混淆植物。
同时本研究建立了一个UPLC-PDA法,实现对市售和实地采集的虎杖药材(38批次)5个主要成分的含量测定,通过数据统计分析揭示该药材质量的控制的关键指标,结合中药特征图谱对38批样品进行内在质量的综合评价和整体物质的全面控制；建立了特征图谱共有模式,并通过聚类分析和主成分分析对虎杖进行了初步评价。结果表明陕西、湖北和河南等地的22批样品仅大黄素甲醚含量较高；湖南和广西等地7批样品的白藜芦醇含量稍高,而虎杖苷含量明显较低；安徽、江苏等地的9批样品虎杖苷和大黄素含量均较高。
本文应用HPLC-DAD-MS"技术对虎杖叶中主要成分进行了鉴定,共推断了15个化合物。并对9批不同产地虎杖叶的8个主要成分进行了含量测定。采用DPPH-HPLC和lipid peroxidation-HPLC两种方法快速鉴定了虎杖叶中的主要抗氧化成分。并测试了各产地样品和主要抗氧化成分的自由基清除能力。结合含量测定的研究结果将样品的活性与产地、采收时间的关系进行了探讨。
Polygonum cuspidatum Sieb. et Zucc., distributed mainly in temperate Asia, are thickets in valleys, field margins, mountain slopes in many areas in China. P. cuspidatum has been used in many Asian countries as traditional medicinal herbs for centuries. The root and radix of P. cuspidatum is a well-known Chinese herbal medicine, and nowadays it is recorded in the Pharmacopaeia of the People's Republic of China. It is traditionally used for dispelling wind-evil, damp elimination, eliminating stasis to active blood circulation, analgesic therapy, relieving cough and reducing sputum. P. cuspidatum leaves are traditionally used for the treatment of rheumatism, joint pain, snakebites and skin disease. It is recorded in "Chiu Huang Pen Ts'ao" as an edible wild herb and is a popular edible wild herb in many areas in China, such as Fujian, Jiangxi, Yunnan, and Henan province.
In the present study, the preliminary pharmacological study of major compounds, the identification of P. cuspidatum and its adulterants, the quality control method of root and radix of P. cuspidatum were studied. Meanwhile, P. cuspidatum leaves were also studied for further development:the identification of chemical constituents, the content determination, and the antioxidant activities.
In vitro activities of main components were studied. Hydroxyanthraquinones in P. cuspidatum, such as emodin, physcion, citreorosein, aloeemodin, chrysophanol, and rhein were main bioactive compounds for improving glucose uptake in HepG2cell. However, their glycosides showed no activities. Therefore, their hydroxyl groups were deduced to be essential for activities. Compounds with different structure types in P. cuspidatum showed inhibition effects against lipid accumulation. They are polydation, hyperin, emodin-8-O-glucoside, rhein, and aloeemodin.
The psbA-trnH sequence was used as DNA Barcode for distinguish twenty four samples of P. cuspidatum (roots:14, leaves:10) and their adulterants. Intraspecific variation was analyzed. P. cuspidatum samples and other Polygonaceae herbs were discriminated using BLAST, nearest distance, and NJ tree methods. There were a low degree of intraspecific variation, and a relatively high degree of interspecific variation for successful species identification.
A UPLC-PDA method was developed for simultaneous quantitative determination of5compounds in38root and radix samples of P. cuspidatum collected from markets or habitats. The UPLC fingerprint was also used for quality control. Common pattern of chromatograph was established, and similarity analysis, hierarchical cluster analysis and principal component analysis were applied for further evaluation. Physcion contents in22samples form Shaanxi, Hubei and Henan were relatively higher; resveratrol contents in7samples from Hunan and Guangxi were relatively higher, while polydatin contents were obviously lower; polydatin and emodin contents in9samples from Anhui and Jiangsu were relatively higher.
The HPLC-DAD-MSn method was developed to study the major components from P. cuspidatum leaves. Twenty-eight compounds were separated, and15of them were tentatively characterized based on their UV and MS spectra. Eight compounds were further ascertained by comparison with the corresponding reference compounds, and quantified in9batches of leaf samples. The antioxidants were screened by DPPH-HPLC and lipid peroxidation-HPLC method. The antioxidant abilities were evaluated for the leaves and compounds as well. The possible relationships among the compound content, antioxidant abilities, collection time, and resource regions were discussed.
对虎杖根的提取物及主要单体化合物进行了降糖降脂活性的体外实验。结果表明虎杖根中促进细胞葡萄糖摄取的成分主要为乙酸乙酯萃取物中的蒽醌类成分。羟基蒽醌类化合物大黄素、大黄素甲醚、6-羟基芦荟大黄素、芦荟大黄素、大黄酚、大黄酸为主要的活性成分；而羟基蒽醌的糖苷类化合物并无明显改善活性,因而初步推断蒽醌类化合物的游离羟基是促进细胞葡萄糖摄取的必要条件。虎杖根中抑制脂质堆积的活性成分主要集中在正丁醇部位,虎杖苷、金丝桃苷、大黄素-8-O-葡萄糖苷、大黄酸和芦荟大黄素为主要的活性成分。
本研究从分子水平应用叶绿体psbA-trnH基因对不同产地采集的24份虎杖样品(包括根14份、叶10份)和同科易混淆植物32条序列进行DNA的分子鉴定研究。对虎杖样品的种内变异情况做了统计,并采用了BLAST、最小距离法、NJ邻接法构建系统发育树来区分虎杖和同科易混淆植物。结果显示虎杖种内最小变异为0,最大变异为7个碱基变异及2个碱基的插入缺失,表明虎杖种内不同居群材料在分子水平上差异较小。而虎杖与同属及同科易混淆植物在分子水平上差异较大。证实psbA-trnH条形码序列能够准确鉴别虎杖药材的基原植物,同时也能准确区分虎杖药材与同科易混淆植物。
同时本研究建立了一个UPLC-PDA法,实现对市售和实地采集的虎杖药材(38批次)5个主要成分的含量测定,通过数据统计分析揭示该药材质量的控制的关键指标,结合中药特征图谱对38批样品进行内在质量的综合评价和整体物质的全面控制；建立了特征图谱共有模式,并通过聚类分析和主成分分析对虎杖进行了初步评价。结果表明陕西、湖北和河南等地的22批样品仅大黄素甲醚含量较高；湖南和广西等地7批样品的白藜芦醇含量稍高,而虎杖苷含量明显较低；安徽、江苏等地的9批样品虎杖苷和大黄素含量均较高。
本文应用HPLC-DAD-MS"技术对虎杖叶中主要成分进行了鉴定,共推断了15个化合物。并对9批不同产地虎杖叶的8个主要成分进行了含量测定。采用DPPH-HPLC和lipid peroxidation-HPLC两种方法快速鉴定了虎杖叶中的主要抗氧化成分。并测试了各产地样品和主要抗氧化成分的自由基清除能力。结合含量测定的研究结果将样品的活性与产地、采收时间的关系进行了探讨。
Polygonum cuspidatum Sieb. et Zucc., distributed mainly in temperate Asia, are thickets in valleys, field margins, mountain slopes in many areas in China. P. cuspidatum has been used in many Asian countries as traditional medicinal herbs for centuries. The root and radix of P. cuspidatum is a well-known Chinese herbal medicine, and nowadays it is recorded in the Pharmacopaeia of the People's Republic of China. It is traditionally used for dispelling wind-evil, damp elimination, eliminating stasis to active blood circulation, analgesic therapy, relieving cough and reducing sputum. P. cuspidatum leaves are traditionally used for the treatment of rheumatism, joint pain, snakebites and skin disease. It is recorded in "Chiu Huang Pen Ts'ao" as an edible wild herb and is a popular edible wild herb in many areas in China, such as Fujian, Jiangxi, Yunnan, and Henan province.
In the present study, the preliminary pharmacological study of major compounds, the identification of P. cuspidatum and its adulterants, the quality control method of root and radix of P. cuspidatum were studied. Meanwhile, P. cuspidatum leaves were also studied for further development:the identification of chemical constituents, the content determination, and the antioxidant activities.
In vitro activities of main components were studied. Hydroxyanthraquinones in P. cuspidatum, such as emodin, physcion, citreorosein, aloeemodin, chrysophanol, and rhein were main bioactive compounds for improving glucose uptake in HepG2cell. However, their glycosides showed no activities. Therefore, their hydroxyl groups were deduced to be essential for activities. Compounds with different structure types in P. cuspidatum showed inhibition effects against lipid accumulation. They are polydation, hyperin, emodin-8-O-glucoside, rhein, and aloeemodin.
The psbA-trnH sequence was used as DNA Barcode for distinguish twenty four samples of P. cuspidatum (roots:14, leaves:10) and their adulterants. Intraspecific variation was analyzed. P. cuspidatum samples and other Polygonaceae herbs were discriminated using BLAST, nearest distance, and NJ tree methods. There were a low degree of intraspecific variation, and a relatively high degree of interspecific variation for successful species identification.
A UPLC-PDA method was developed for simultaneous quantitative determination of5compounds in38root and radix samples of P. cuspidatum collected from markets or habitats. The UPLC fingerprint was also used for quality control. Common pattern of chromatograph was established, and similarity analysis, hierarchical cluster analysis and principal component analysis were applied for further evaluation. Physcion contents in22samples form Shaanxi, Hubei and Henan were relatively higher; resveratrol contents in7samples from Hunan and Guangxi were relatively higher, while polydatin contents were obviously lower; polydatin and emodin contents in9samples from Anhui and Jiangsu were relatively higher.
The HPLC-DAD-MSn method was developed to study the major components from P. cuspidatum leaves. Twenty-eight compounds were separated, and15of them were tentatively characterized based on their UV and MS spectra. Eight compounds were further ascertained by comparison with the corresponding reference compounds, and quantified in9batches of leaf samples. The antioxidants were screened by DPPH-HPLC and lipid peroxidation-HPLC method. The antioxidant abilities were evaluated for the leaves and compounds as well. The possible relationships among the compound content, antioxidant abilities, collection time, and resource regions were discussed.
引文
[1]吴征镒,路安民,汤彦承,等.中国被子植物科属综论[M].北京：科学出版社,2003.164-167.
[2]李安仁.中国植物志[A].北京：科学出版社,1998.1.
[3]Li A, Bao B, Alisan E G-B, et al. Polygonaceae in Wu Z Y and Raven P H(editors), Flora of China[M]. Beijing:Science Press, St.Louis:Missouri Botanical Garden Press. 2003,5:277-350.
[4]Beerling D J, Bailey J P, Conolly A P. Fallopia japonica (Houtt.) Ronse Decraene[J]. Journal of Ecology,1994:959-979.
[5]Holub J. Fallopia ADANS.1763 instead of Bilderdykia DUM.1827[J]. Folia Geobotanica,1971,6(2):171-177.
[6]Kim J Y, Park C. Morphological and chromosomal variation in Fallopia section Reynoutria (Polygonaceae) in Korea[J]. Brittonia,2000,52(1):34-48.
[7]Mandak B, Bimova K, Pysek P, et al. Isoenzyme diversity in Reynoutria (Polygonaceae) taxa:escape from sterility by hybridization[J]. Plant Systematics and Evolution,2005,253(1):219-230.
[8]Bailey J P, Stace C A. Chromosome number, morphology, pairing, and DNA values of species and hybrids in the genus Fallopia (Polygonaceae)[J]. Plant Systematics and Evolution,1992,180(1):29-52.
[9]Decraene L P R, Akeroyd J R. Generic limits in Polygonum and related genera (Polygonaceae) on the basis of floral characters[J]. Botanical Journal of the Linnean Society,1988,98(4):321-371.
[10]Lamb Frye A S, Kron K A. rbcL phylogeny and character evolution in Polygonaceae[J]. Systematic Botany,2003,28(2):326-332.
[11]Bailey J P, Bimova K, Mandak B. The potential role of polyploidy and hybridisation in the further evolution of the highly invasive Fallopia taxa in Europe[J]. Ecological Research,2007,22(6):920-928.
[12]刘明珍.基于atpB-rbcL基因序列及茎的解剖结构特征对蓼科部分属间关系的研究[D].安徽大学,2007.
[13]曲畅游.中国蓼族植物果实形态,果皮微形态研究及其于trnk基因5’端内含子序列分析探讨虎杖的系统学位置[D].山东师范大学,2005.
[14]马云桐.虎杖的资源、品质与药效的相关性研究[D].成都中医药大学,2006.10.
[15]Chinese Pharmacopoeia Commission. Pharmacopoeia of the People's Republic of China (Vol.1,2010 Edition)[M]. Beijing:China Medical Science and Technology Press, 2010.194.
[16]国家中医药管理局<中华本草>编委会.中华本草[A].上海：上海科技出版社,1999.659.
[17]中国药材公司.中国中药资源志要[M].北京：科学出版社,1994.223.
[18]Nonomura S, Kanagawa H, Makimoto A. Chemical constituents of polygonaceous plants. I. Components of Polygonum cuspidatum[J]. Yakugaku zasshi:Journal of the Pharmaceutical Society of Japan,1963,83:988-990.
[19]Jayatilake G S, Jayasuriya H, Lee E, et al. Kinase inhibitors from Polygonum cuspidatum[J]. Journal of Natural Products,1993,56(10):1805-1810.
[20]Vastano B C, Chen Y, Zhu N, et al. Isolation and identification of stilbenes in two varieties of Polygonum cuspidatum[J]. Journal of Agricultural and Food Chemistry,2000, 48(2):253-256.
[21]Hegde V R, Pu H, Patel M, et al. Two new bacterial DNA primase inhibitors from the plant Polygonum cuspidatum [J]. Bioorganic & Medicinal Chemistry Letters,2004, 14(9):2275-2277.
[22]Xiao K, Xuan L, Xu Y, et al. Stilbene glycoside sulfates from Polygonum cuspidatum[J]. Journal of Natural Products,2000,63(10):1373-1376.
[23]Xiao K, Xuan L, Xu Y, et al. Constituents from Polygonum cuspidatum[J]. Chemical and Pharmaceutical Bulletin,2002,50(5):605-608.
[24]Xiao K, Xuan L, Xu Y, et al. Dimeric stilbene glycosides from Polygonum cuspidatum[J]. European Journal of Organic Chemistry,2002,2002(3):564-568.
[25]Tsukida K, Michikoyoneshige. Studies on the constituents of Polygonaeeous plants. III. constituents of Ko-jo-kon. (Polygonum cuspidatum sieb. et Zucc).[J].Yakugaku zasshi:Journal of the Pharmaceutical Society of Japan,1954,74(4):379-382.
[26]朱廷儒,王素贤,裴月湖.中药虎杖抗菌活性成分的研究[J].中草药,1985,10(3)：21.
[27]Kimura Y, Kozawa M, Baba K, et al. New constituents of roots of Polygonum cuspidatum[J]. Planta Medica,1983,48(07):164-168.
[28]金雪梅,金光洙.虎杖的化学成分研究[J].中草药,2007,38(10)：1446-1448.
[29]Kuznetsova Z P. Study of phenolic compounds from Japanese knotweed (Polygonum cuspidatum)[J].Vestsi Akad Navuk Belarus SSR Ser Biyal Navuk,1979,5: 29.
[30]Molnar B. Phytochemical examination of the active polyphenolic ingredients of the Japanese bitter herb (Reynoutria japonica Houtt.)[J]. Gyogyszereszet,1991,35(1): 47-52.
[31]肖凯,宣利江,徐亚明,等.虎杖的化学成分研究[J].中国药学杂志,2003,38(1)：12-15.
[32]Rho T C, Choi H C, Lee S W, et al.. Inhibition of nitric oxide synthesis by coumarins from Polygonum cuspidatum in LPS-Activated RAW 264.7 cells[J]. Korean Journal of Pharmacognosy,2001,32(3):181-188.
[33]肖凯,宣利江,徐亚明,等.虎杖的水溶性成分研究[J].中草药,2003,34(6)：496-498.
[34]Vechar A, Kuznyetsova Z, Chekalinskaya I. Study of phenolcarboxylic acids of Polygonum species[J]. Vestsi Akademii Navuk BSSR, Seryya Biyalagichnykh Navuk, 1980, (6):71-74.
[35]Kim T, Hee C, Yang K, et al. Studies on the constituents of Polygonum sp.[J]. Saengyak Hakhoechi,1979,9(2):89-91.
[36]Kim Y S, Hwang C S, Shin D H. Volatile constituents from the leaves of Polygonum cuspidatum S. et Z. and their anti-bacterial activities[J]. Food Microbiology, 2005,22(1):139-144.
[37]Hemberg S. Tanniferous kinds of Polygonum[J]. Leder,1951, (2):239-241.
[38]Li F, Zhan Z, Liu F, et al. Polyflavanostilbene A, a new flavanol-fused stilbene glycoside from Polygonum cuspidatum[J]. Organic Letters,2013,15(3):674-677.
[39]Zhang H, Zhang Q, Wang L, et al. Two new anthraquinone malonylglucosides from Polygonum cuspidatum[J]. Natural Product Research,2012,26(14):1323-1327.
[40]张彩宁.中药虎杖雌激素活性成分及其物质基础研究[D].中国科学院研究生院,2006.
[41]Kuznetsova Z P. Study of phenolic compounds from Japanese knotweed (Polygonum cuspidatum)[J]. Vestsi Akademii Navuk BSSR, Seryya Biyalagichnykh Navuk,1979:29-32.
[42]俸灵林.虎杖的化学成分及其质量研究[D].沈阳：沈阳药科大学,2003.
[43]Liu X, Wu L, Song G, et al. Study on chemical constituents of Polygonum cuspidatum Sieb[J]. Shenyang Yaoke Daxue Xuebao,1999,16(Suppl.):17-20.
[44]刘晓秋,于黎明,吴立军.虎杖化学成分研究(Ⅰ)[J].中国中药杂志,2003,28(1)：47-49.
[45]张海防,窦昌贵,刘晓华,等.虎杖提取物抗炎作用的实验研究[J].药学进展,2003,27(4)：230-233.
[46]Bralley E E, Greenspan P, Hargrove J L, et al. Topical anti-inflammatory activity of Polygonum cuspidatum extract in the TPA model of mouse ear inflammation[J]. Journal of Inflammation,2008,5(1):1.
[47]Banerjee S, Bueso-Ramos C, Aggarwal B B. Suppression of 7,12-Dimethylbenz (a) anthracene-induced mammary carcinogenesis in rats by resveratrol role of nuclear factor-κB, cyclooxygenase 2, and matrix metalloprotease 9[J]. Cancer Research,2002, 62(17):4945-4954.
[48]Manna S K, Mukhopadhyay A, Aggarwal B B. Resveratrol suppresses TNF-induced activation of nuclear transcription factors NF-κB, activator protein-1, and apoptosis:potential role of reactive oxygen intermediates and lipid peroxidation[J]. The Journal of Immunology,2000,164(12):6509-6519.
[49]Kumar A, Dhawan S, Aggarwal B B. Emodin (3-methyl-1,6, 8-trihydroxyanthraquinone) inhibits TNF-induced NF-kappaB activation, IkappaB degradation, and expression of cell surface adhesion proteins in human vascular endothelial cells.[J]. Oncogene,1998,17(7):913.
[50]Rhotc, He C, Sw L, et al. Inhibition of nitric oxide synthesis by coumarins from 264.7 cells Polygonum cuspidatum in LPS-activated RAW[J]. Saengyak Hakhoechi, 2001,32(3):181-188.
[51]Kimura Y, Okuda H, Kubo M. Effects of stilbenes isolated from medicinal plants on arachidonate metabolism and degranulation in human polymorphonuclear leukocytes[J]. Journal of Ethnopharmacology,1995,45(2):131-139.
[52]Shan B, Cai Y, Brooks J D, et al. Antibacterial properties of Polygonum cuspidatum roots and their major bioactive constituents[J]. Food Chemistry,2008,109(3): 530-537.
[53]Kukric Z Z, Topalic-Trivunovic L N. Antibacterial activity of cis-and trans-resveratrol isolated from Polygonum cuspidatum rhizome[J]. Acta Periodica Technologica 2006, (37):131-136.
[54]Song J, Yang T, Chang K, et al. In vitro effects of a fraction separated from Polygonum cuspidatum root on the viability, in suspension and biofilms, and biofilm formation of mutans streptococci [J]. Journal of Ethnopharmacology,2007,112(3): 419-425.
[55]K Y. Antimicrobial activity in natural resources. I. Antifungal and fungicidal activities of catechin and crude drugs against Trichophyton[J]. Toyama-ken Yakuji Kenkyusho Nenpo,1992,19:82-87.
[56]Lin H, Sun M, Wang Y, et al. Anti-HIV activities of the compounds isolated from Polygonum cuspidatum and Polygonum multiflorum[J]. Planta Medica,2010,76(09): 889-892.
[57]Y J, Hx W, Zy B, et al. Evaluation of antiviral effect of Polygonum cuspidatum water extract with a model of murine acquired immunodeficiency syndrome[J]. Virologica Sinica,1998,13(4):306-311.
[58]李凤新,孙振国,苏红,等.虎杖蒽醌提取物抗柯萨奇B3型病毒作用及机制研究[J].中草药,2008,39(11)：1699-1701.
[59]关文达,杨子峰,刘妮,等.白藜芦醇对多种呼吸道病毒作用体外实验研究[J].中药材,2008,31(9)：1388-1390.
[60]Chang J, Liu H, Wang K, et al. Ethanol extract of Polygonum cuspidatum inhibits hepatitis B virus in a stable HBV-producing cell line[J]. Antiviral Research,2005,66(1): 29-34.
[61]Park C, Lee Y, Kim J, et al. Inhibitory effects of Polygonum cuspidatum water extract (PCWE) and its component rasveratrol on acyl-coenzyme A-cholesterol acyltransferase activity for cholesteryl ester synthesis in HepG2 cells[J]. Vascular Pharmacology,2004,40(6):279-284.
[62]Xing W, Wu J, Jia M, et al. Effects of polydatin from Polygonum cuspidatum on lipid profile in hyperlipidemic rabbits[J]. Biomedicine & Pharmacotherapy,2009,63(7): 457-462.
[63]Du J, Sun L, Xing W, et al. Lipid-lowering effects of polydatin from Polygonum cuspidatum in hyperlipidemic hamsters[J]. Phytomedicine,2009,16(6):652-658.
[64]Sagar S M, Yance D, Wong R K. Natural health products that inhibit angiogenesis: a potential source for investigational new agents to treat cancer-Part 1[J]. Current Oncology,2006,13(1):14.
[65]Kimura Y, Okuda H. Resveratrol isolated from Polygonum cuspidatum root prevents tumor growth and metastasis to lung and tumor-induced neovascularization in Lewis lung carcinoma-bearing mice[J]. The Journal of Nutrition,2001,131(6): 1844-1849.
[66]Hsu C, Chan Y, Chang J. Antioxidant activity of extract from Polygonum cuspidatum[J]. Biological Research,2007,40(1):13-21.
[67]Pan Y, Zhang X, Wang H, et al. Antioxidant potential of ethanolic extract of Polygonum cuspidatum and application in peanut oil[J]. Food Chemistry,2007,105(4): 1518-1524.
[68]Zhang C Z, Wang S X, Zhang Y, et al. In vitro estrogenic activities of Chinese medicinal plants traditionally used for the management of menopausal symptoms[J]. Journal of Ethnopharmacology,2005,98(3):295-300.
[69]Zhang C, Wang X, Zhang X, et al. Bioassay-guided separation of citreorosein and other oestrogenic compounds From Polygonum cuspidatum[J]. Phytotherapy Research, 2009,23(5):740-741.
[70]Matsuda H, Shimoda H, Morikawa T, et al. Phytoestrogens from the roots of Polygonum cuspidatum (Polygonaceae):structure-requirement of hydroxyanthraquinones for estrogenic activity[J]. Bioorganic & Medicinal Chemistry Letters,2001,11(14): 1839-1842.
[71]Liu Z P, Li W X, Yu B, et al. Effects of trans-resveratrol from Polygonum cuspidatum on bone loss using the ovariectomized rat model [J]. Journal of Medicinal Food,2005,8(1):14-19.
[72]蔡宝昌,刘训红.常用中药材HPLC指纹图谱测定技术[M].北京：化学工业出版社,2005.2.
[73]易伦朝,吴海,梁逸曾.色谱指纹图谱与中药质量控制[J].色谱,2008,26(2)：166-171.
[74]谢培山.色谱指纹图谱分析是中草药质量控制的可行策略[J].中药新药与临床药理,2001,12(3)：141-151.
[75]彭静波,李强,雷海民,等.虎杖的功效成分组研究Ⅰ——延长凝血时间作用的谱效分析研究[J].中华中医药杂志,2011,26(8)：1737-1739.
[76]彭静波,李强,司银楚,等.虎杖的功效成分组研究Ⅱ——对全血黏度影响的谱效分析研究[J].中华中医药杂志,2011,26(9)：1964-1966.
[77]龙海林,李强,司银楚,等.虎杖的功效成分组研究Ⅲ——对血浆黏度影响的谱效分析研究[J].现代医药卫生,2012,28(020)：3080-3082.
[78]龙海林,雷海民,李强,等.虎杖的功效成分组研究Ⅳ-纤维蛋白原谱效分析的研究[J].中国医药指南,2012,10(33)：4-5.
[79]李强,夏晓晖,雷海民,等.虎杖功效成分组研究V-血沉沉降速率谱效关系研究[J].中医药学报,2011,39(5)：63.
[80]陈华国,赵宏宾,赵超,等.虎杖提取物HPLC指纹图谱归属分析及谱效关 系初探[J].中国药房,2010,(19)：1775-1776.
[81]罗志江,徐彦,吴建英,等.虎杖指纹图谱及其抗氧化活性的“谱-效”关系研究[J].西南大学学报(自然科学版),2012,34(1)：138-142.
[82]Zuo H, Shi Z, Hu X, et al. Prevalence of metabolic syndrome and factors associated with its components in Chinese adults[J]. Metabolism, Clinical and Experimental,2009,58(8):1102-1108.
[83]Alfaradhi M Z, Ozanne S E. Developmental programming in response to maternal overnutrition[J]. Frontiers in Genetics,2011,2,27.
[84]Despres J, Lemieux I, Bergeron J, et al. Abdominal obesity and the metabolic syndrome:contribution to global cardiometabolic risk[J]. Arteriosclerosis, Thrombosis, and Vascular Biology,2008,28(6):1039-1049.
[85]Dandona P, Aljada A, Bandyopadhyay A. Inflammation:the link between insulin resistance, obesity and diabetes[J]. Trends in Immunology,2004,25(1):4-7.
[86]Bradbury M W. Lipid metabolism and liver inflammation. I. Hepatic fatty acid uptake:possible role in steatosis[J]. American Journal of Physiology-Gastrointestinal and Liver Physiology,2006,290(2):G194-G198.
[87]Rector R S, Thyfault J P, Wei Y, et al. Non-alcoholic fatty liver disease and the metabolic syndrome:an update[J]. World Journal of Gastroenterology:WJG,2008,14(2): 185.
[88]Festi D, Colecchia A, Sacco T, et al. Hepatic steatosis in obese patients:clinical aspects and prognostic significance[J]. Obesity Reviews,2004,5(1):27-42.
[89]Unger R H. Lipid overload and overflow:metabolic trauma and the metabolic syndrome[J]. Trends in Endocrinology & Metabolism,2003,14(9):398-403.
[90]de Luca C, Olefsky J M. Stressed out about obesity and insulin resistance [J]. Nature Medicine,2006,12(1):41-42.
[91]Morino K, Petersen K F, Shulman G I. Molecular mechanisms of insulin resistance in humans and their potential links with mitochondrial dysfunction[J]. Diabetes,2006, 55(Suppl.2):S9-S15.
[92]Schinner S, Scherbaum W A, Bornstein S R, et al. Molecular mechanisms of insulin resistance[J]. Diabetic Medicine,2005,22(6):674-682.
[93]Steinberger J, Daniels S R. Obesity, Insulin Resistance, Diabetes, and Cardiovascular Risk in Children An American Heart Association Scientific Statement From the Atherosclerosis, Hypertension, and Obesity in the Young Committee (Council on Cardiovascular Disease in the Young) and the Diabetes Committee (Council on Nutrition, Physical Activity, and Metabolism)[J]. Circulation,2003,107(10):1448-1453.
[94]Cohen J C, Horton J D, Hobbs H H. Human fatty liver disease:old questions and new insights[J]. Science,2011,332(6037):1519-1523.
[95]Starley B Q, Calcagno C J, Harrison S A. Nonalcoholic fatty liver disease and hepatocellular carcinoma:a weighty connection[J]. Hepatology,2010,51(5):1820-1832.
[96]Ahmed M H, Byrne C D. Current treatment of non-alcoholic fatty liver disease[J]. Diabetes, Obesity and Metabolism,2009,11(3):188-195.
[97]Ioannides-Demos L L, Proietto J, Tonkin A M, et al. Safety of drug therapies used for weight loss and treatment of obesity[J]. Drug Safety,2006,29(4):277-302.
[98]Vermaak I, Viljoen A M, Hamman J H. Natural products in anti-obesity therapy[J]. Natural Product Reports,2011,28(9):1493-1533.
[99]王辉,杨再刚,燕树勋,等.虎杖总蒽醌对糖尿病肾病早期血瘀模型大鼠脂代谢及血液流变性的影响[J].中国实验方剂学杂志,2010,16(16).
[100]郑晓媛.虎杖提取物对α-葡萄糖苷酶的抑制作用研究[D].四川大学,2007.
[101]沈忠明,殷建伟,袁海波.虎杖鞣质的降血糖作用研究[J].天然产物研究与开发,2004,16(3).
[102]Ricchi M, Odoardi M R, Carulli L, et al. Differential effect of oleic and palmitic acid on lipid accumulation and apoptosis in cultured hepatocytes[J]. Journal of Gastroenterology and Hepatology,2009,24(5):830-840.
[103]Morikawa T, Ninomiya K, Zhang Y, et al. Flavonol glycosides with lipid accumulation inhibitory activity from Sedum sarmentosum[J].Phytochemistry Letters, 2012,5(1):53-58.
[104]方飞,罗明俐,苏楠,等.桑叶提取物对胰岛素抵抗HepG2细胞葡萄糖摄取的影响及其分子机制[J].药学学报,2012,47(011)：1452-1456.
[105]杨桂枝,高小平,晏菊芳,等.GOD-POD法微量化测定方法的建立及其在3T3-L1脂肪细胞和HepG2细胞糖摄取中的应用[J].四川解剖学杂志,2003：12-15.
[106]姜斐,姚楠,钱士辉,等.女贞子中8个化合物对胰岛素抵抗HepG2细胞葡萄糖消耗的影响[J].海峡药学,2010,(001)：164-167.
[107]Huang Y, Lin C, Huang S, et al. Effect and mechanism of ginsenosides CK. and Rgl on stimulation of glucose uptake in 3T3-L1 adipocytes[J]. Journal of Agricultural and Food Chemistry,2010,58(10):6039-6047.
[108]Ann Y M, Kim S K, Kang J S, et al. Platycodon grandiflorum modifies adipokines and the glucose uptake in high-fat diet in mice and L6 muscle cells[J]. Journal of Pharmacy and Pharmacology,2012,64(5):697-704.
[109]Noh J, Lee I, Ly S, et al. A Phellinus baumii extract reduces obesity in high-fat diet-fed mice and absorption of triglyceride in lipid-loaded mice[J]. Journal of Medicinal Food,2011,14(3):209-218.
[110]Zou C, Wang Y, Shen Z.2-NBDG as a fluorescent indicator for direct glucose uptake measurement[J]. Journal of Biochemical and Biophysical Methods,2005,64(3): 207-215.
[111]李晨钟,张素华,舒昌达,等.胰岛素抵抗细胞模型的胰岛素降解酶活性与胰岛素敏感性的关系[J].第一军医大学学报,2002,22(2)：151-154.
[112]李兰芳,黄秀清,原慧萍,等.吡格列酮改善胰岛素抵抗状态下氧化应激水平及分子机制探讨[J].中国临床药理学与治疗学,2009,14(3)：245-249.
[113]李美妮,韩蕊莲,韩建萍,等.大黄与易混伪品土大黄,虎杖原植物的ITS2序列鉴定[J].环球中医药,2012,3：10.
[114]Frederich M, Wauters J N, Tits M, et al. Quality Assessment of Polygonum cuspidatum and Polygonum multiflorum by 1H NMR metabolite fingerprinting and profiling analysis[J]. Planta Medica,2011,77:81-86.
[115]Qian G, Leung S, Lu G, et al. Differentiation of Rhizoma et Radix Polygoni cuspidati from closely related herbs by HPLC fingerprinting[J]. Chemical and Pharmaceutical Bulletin,2006,54(8):1179-1186.
[116]Hebert P D, Cywinska A, Ball S L. Biological identifications through DNA barcodes[J]. Proceedings of the Royal Society of London. Series B:Biological Sciences, 2003,270(1512):313-321.
[117]Gregory T R. DNA barcoding does not compete with taxonomy[J]. Nature,2005, 434(7037):1067.
[118]Miller S E. DNA barcoding and the renaissance of taxonomy [J]. Proceedings of the National Academy of Sciences,2007,104(12):4775-4776.
[119]陈士林,宋经元,姚辉.药用植物DNA条形码鉴定策略及关键技术分析[J].中国天然药物,2009,7(5)：322-327.
[120]陈士林,姚辉,宋经元,等.基于DNA barcoding(条形码)技术的中药材鉴定[J].世界科学技术：中医药现代化,2007,9(3)：7-12.
[121]罗焜,陈科力,刘义梅,等.高良姜及其混淆品的分子鉴定[J].世界科学技术：中医药现代化,2011,13(2)：400-406.
[122]罗焜,陈士林,陈科力,等.基于芸香科的植物通用DNA条形码研究[J].中国科学：生命科学,2010,(4)：342-351.
[123]Pang X, Song J, Zhu Y, et al. Applying plant DNA barcodes for Rosaceae species identification[J]. Cladistics,2011,27(2):165-170.
[124]Gao T, Yao H, Song J, et al. Identification of medicinal plants in the family Fabaceae using a potential DNA barcode ITS2[J]. Journal of Ethnopharmacology,2010, 130(1):116-121.
[125]朱英杰,陈士林,姚辉,等.重楼属药用植物DNA条形码鉴定研究[J].药学学报,2010,45(3)：376-382.
[126]Yao H, Song J, Liu C, et al. Use of ITS2 region as the universal DNA barcode for plants and animals[J]. PloS One,2010,5(10):e13102.
[127]Kress W J, Erickson D L. A two-locus global DNA barcode for land plants:the coding rbcL gene complements the non-coding trnH-psbA spacer region[J], PLoS One, 2007,2(6):e508.
[128]Lahaye R, van der Bank M, Bogarin D, et al. DNA barcoding the floras of biodiversity hotspots[J]. Proceedings of the National Academy of Sciences,2008,105(8): 2923-2928.
[129]Chase M W, Cowan R S, Hollingsworth P M, et al. A proposal for a standardised protocol to barcode all land plants[J]. Taxon,2007:295-299.
[130]Chen S, Yao H, Han J, et al. Validation of the ITS2 region as a novel DNA barcode for identifying medicinal plant species[J]. PloS one,2010,5(1):e8613.
[131]Hollingsworth P M, Forrest L L, Spouge J L, et al. A DNA barcode for land plants[J]. Proceedings of the National Academy of Sciences,2009,106(31): 12794-12797.
[132]Kress W J, Wurdack K J, Zimmer E A, et al. Use of DNA barcodes to identify flowering plants[J]. Proceedings of the National Academy of Sciences of the United States of America,2005,102(23):8369-8374.
[133]Chase M W, Salamin N, Wilkinson M, et al. Land plants and DNA barcodes: short-term and long-term goals[J]. Philosophical Transactions of the Royal Society B: Biological Sciences,2005,360(1462):1889-1895.
[134]Gonzalez M A, Baraloto C, Engel J, et al. Identification of Amazonian trees with DNA barcodes[J]. PLoS One,2009,4(10):e7483.
[135]E P. Wanted:a barcode for plants[J]. Science,2007,318:190-191.
[136]Keller A, Schleicher T, Schultz J, et al.5.8 S-28S rRNA interaction and HMM-based ITS2 annotation.[J]. Gene,2009,430(1-2):50.
[137]Muller T, Philippi N, Dandekar T, et al. Distinguishing species[J]. RNA,2007, 13(9):1469-1472.
[138]Miao M, Warren A, Song W, et al. Analysis of the internal transcribed spacer 2 (ITS2) region of scuticociliates and related taxa (Ciliophora, Oligohymenophorea) to infer their evolution and phylogeny[J]. Protist,2008,159(4):519-533.
[139]Shaw J, Lickey E B, Beck J T, et al. The tortoise and the hare Ⅱ:relative utility of 21 noncoding chloroplast DNA sequences for phylogenetic analysis[J]. American Journal of Botany,2005,92(1):142-166.
[140]Yao H, Song J, Ma X, et al. Identification of Dendrobium species by a candidate DNA barcode sequence:the chloroplast psbA-trnH intergenic region[J]. Planta Medica, 2009,75(06):667-669.
[141]Zhao Z, Xiao P. Encyclopedia of medicinal plants (Volume II)[M]. Shanghai: World Publishing Corporation,2007.235-239.
[142]马云桐,万德光,宋良科.虎杖药材HPLC指纹图谱研究[J].中国中药杂志,2006,31(12)：972-974.
[143]雷海民,毕葳,李强,等.虎杖药材指纹图谱研究[J].中草药,2006,37(7)：1084-1086.
[144]张明昶,彭小冰,苗旭辉,等.中药虎杖特征指纹图谱研究[J].中国实验方剂学杂志,2011,17(1)：53.
[145]李宇翔,王丽,唐书明,等.虎杖HPLC指纹图谱研究与应用[J].中国实验方剂学杂志,2010,16(5)：78.
[146]马培,许利嘉,刘延泽,等.虎杖药材UPLC特征图谱研究[J].中国实验方剂学杂志,2012,8(2).
[147]Ma P, Luo K, Peng Y, et al. Quality control of Polygonum cuspidatum by UPLC-PDA and related statistical analysis[J]. Journal of Liquid Chromatography & Related Technologies, doi:10.1080/10826076.2012.723096.
[148]王玺,王文宇,张克荣,等.中药HPLC指纹图谱相似性研究的探讨[J].沈阳药科大学学报,2003,20(5)：360-362.
[149]孟庆华,刘永锁,蒋淑敏,等.色谱指纹图谱综合信息指数在中药质量控制中的应用研究[J].中国天然药物,2004,2(6)：359-364.
[150]Xie G X, Ni Y, Su M M, et al. Application of ultra-performance LC-TOF MS metabolite profiling techniques to the analysis of medicinal Panax herbs[J]. Metabolomics,2008,4(3):248-260.
[151]云南省食品药品监督管理局.云南省中药材标准·第一册[M].昆明：云南美术 出版社,2005.33.
[152]Chandrasekar D, Madhusudhana K, Ramakrishna S, et al. Determination of DPPH free radical scavenging activity by reversed-phase HPLC:A sensitive screening method for polyherbal formulations[J]. Journal of Pharmaceutical and Biomedical Analysis,2006, 40(2):460-464.
[153]Qiu J, Chen L, Zhu Q, et al. Screening natural antioxidants in peanut shell using DPPH-HPLC-DAD-TOF/MS methods[J]. Food Chemistry,2012,135(4):2366-2371.
[154]Tang D, Li H J, Chen J, et al. Rapid and simple method for screening of natural antioxidants from Chinese herb Flos Lonicerae Japonicae by DPPH-HPLC-DAD-TOF/MS[J]. Journal of Separation Science,2008,31(20): 3519-3526.
[155]Shui G, Leong L P. Screening and identification of antioxidants in biological samples using high-performance liquid chromatography-mass spectrometry and its application on Salacca edulis Reinw[J]. Journal of Agricultural and Food Chemistry, 2005,53(4):880-886.
[156]Rice-Evans C A, Miller N J, Bolwell P G, et al. The relative antioxidant activities of plant-derived polyphenolic flavonoids[J]. Free Radical Research,1995,22(4): 375-383.
[157]Vinson J A, Dabbagh Y A, Serry M M, et al. Plant flavonoids, especially tea flavonols, are powerful antioxidants using an in vitro oxidation model for heart disease[J]. Journal of Agricultural and Food Chemistry,1995,43(11):2800-2802.
[158]Laguerre M, Lecomte J, Villeneuve P. Evaluation of the ability of antioxidants to counteract lipid oxidation:existing methods, new trends and challenges[J]. Progress in Lipid Research,2007,46(5):244-282.
[2]李安仁.中国植物志[A].北京：科学出版社,1998.1.
[3]Li A, Bao B, Alisan E G-B, et al. Polygonaceae in Wu Z Y and Raven P H(editors), Flora of China[M]. Beijing:Science Press, St.Louis:Missouri Botanical Garden Press. 2003,5:277-350.
[4]Beerling D J, Bailey J P, Conolly A P. Fallopia japonica (Houtt.) Ronse Decraene[J]. Journal of Ecology,1994:959-979.
[5]Holub J. Fallopia ADANS.1763 instead of Bilderdykia DUM.1827[J]. Folia Geobotanica,1971,6(2):171-177.
[6]Kim J Y, Park C. Morphological and chromosomal variation in Fallopia section Reynoutria (Polygonaceae) in Korea[J]. Brittonia,2000,52(1):34-48.
[7]Mandak B, Bimova K, Pysek P, et al. Isoenzyme diversity in Reynoutria (Polygonaceae) taxa:escape from sterility by hybridization[J]. Plant Systematics and Evolution,2005,253(1):219-230.
[8]Bailey J P, Stace C A. Chromosome number, morphology, pairing, and DNA values of species and hybrids in the genus Fallopia (Polygonaceae)[J]. Plant Systematics and Evolution,1992,180(1):29-52.
[9]Decraene L P R, Akeroyd J R. Generic limits in Polygonum and related genera (Polygonaceae) on the basis of floral characters[J]. Botanical Journal of the Linnean Society,1988,98(4):321-371.
[10]Lamb Frye A S, Kron K A. rbcL phylogeny and character evolution in Polygonaceae[J]. Systematic Botany,2003,28(2):326-332.
[11]Bailey J P, Bimova K, Mandak B. The potential role of polyploidy and hybridisation in the further evolution of the highly invasive Fallopia taxa in Europe[J]. Ecological Research,2007,22(6):920-928.
[12]刘明珍.基于atpB-rbcL基因序列及茎的解剖结构特征对蓼科部分属间关系的研究[D].安徽大学,2007.
[13]曲畅游.中国蓼族植物果实形态,果皮微形态研究及其于trnk基因5’端内含子序列分析探讨虎杖的系统学位置[D].山东师范大学,2005.
[14]马云桐.虎杖的资源、品质与药效的相关性研究[D].成都中医药大学,2006.10.
[15]Chinese Pharmacopoeia Commission. Pharmacopoeia of the People's Republic of China (Vol.1,2010 Edition)[M]. Beijing:China Medical Science and Technology Press, 2010.194.
[16]国家中医药管理局<中华本草>编委会.中华本草[A].上海：上海科技出版社,1999.659.
[17]中国药材公司.中国中药资源志要[M].北京：科学出版社,1994.223.
[18]Nonomura S, Kanagawa H, Makimoto A. Chemical constituents of polygonaceous plants. I. Components of Polygonum cuspidatum[J]. Yakugaku zasshi:Journal of the Pharmaceutical Society of Japan,1963,83:988-990.
[19]Jayatilake G S, Jayasuriya H, Lee E, et al. Kinase inhibitors from Polygonum cuspidatum[J]. Journal of Natural Products,1993,56(10):1805-1810.
[20]Vastano B C, Chen Y, Zhu N, et al. Isolation and identification of stilbenes in two varieties of Polygonum cuspidatum[J]. Journal of Agricultural and Food Chemistry,2000, 48(2):253-256.
[21]Hegde V R, Pu H, Patel M, et al. Two new bacterial DNA primase inhibitors from the plant Polygonum cuspidatum [J]. Bioorganic & Medicinal Chemistry Letters,2004, 14(9):2275-2277.
[22]Xiao K, Xuan L, Xu Y, et al. Stilbene glycoside sulfates from Polygonum cuspidatum[J]. Journal of Natural Products,2000,63(10):1373-1376.
[23]Xiao K, Xuan L, Xu Y, et al. Constituents from Polygonum cuspidatum[J]. Chemical and Pharmaceutical Bulletin,2002,50(5):605-608.
[24]Xiao K, Xuan L, Xu Y, et al. Dimeric stilbene glycosides from Polygonum cuspidatum[J]. European Journal of Organic Chemistry,2002,2002(3):564-568.
[25]Tsukida K, Michikoyoneshige. Studies on the constituents of Polygonaeeous plants. III. constituents of Ko-jo-kon. (Polygonum cuspidatum sieb. et Zucc).[J].Yakugaku zasshi:Journal of the Pharmaceutical Society of Japan,1954,74(4):379-382.
[26]朱廷儒,王素贤,裴月湖.中药虎杖抗菌活性成分的研究[J].中草药,1985,10(3)：21.
[27]Kimura Y, Kozawa M, Baba K, et al. New constituents of roots of Polygonum cuspidatum[J]. Planta Medica,1983,48(07):164-168.
[28]金雪梅,金光洙.虎杖的化学成分研究[J].中草药,2007,38(10)：1446-1448.
[29]Kuznetsova Z P. Study of phenolic compounds from Japanese knotweed (Polygonum cuspidatum)[J].Vestsi Akad Navuk Belarus SSR Ser Biyal Navuk,1979,5: 29.
[30]Molnar B. Phytochemical examination of the active polyphenolic ingredients of the Japanese bitter herb (Reynoutria japonica Houtt.)[J]. Gyogyszereszet,1991,35(1): 47-52.
[31]肖凯,宣利江,徐亚明,等.虎杖的化学成分研究[J].中国药学杂志,2003,38(1)：12-15.
[32]Rho T C, Choi H C, Lee S W, et al.. Inhibition of nitric oxide synthesis by coumarins from Polygonum cuspidatum in LPS-Activated RAW 264.7 cells[J]. Korean Journal of Pharmacognosy,2001,32(3):181-188.
[33]肖凯,宣利江,徐亚明,等.虎杖的水溶性成分研究[J].中草药,2003,34(6)：496-498.
[34]Vechar A, Kuznyetsova Z, Chekalinskaya I. Study of phenolcarboxylic acids of Polygonum species[J]. Vestsi Akademii Navuk BSSR, Seryya Biyalagichnykh Navuk, 1980, (6):71-74.
[35]Kim T, Hee C, Yang K, et al. Studies on the constituents of Polygonum sp.[J]. Saengyak Hakhoechi,1979,9(2):89-91.
[36]Kim Y S, Hwang C S, Shin D H. Volatile constituents from the leaves of Polygonum cuspidatum S. et Z. and their anti-bacterial activities[J]. Food Microbiology, 2005,22(1):139-144.
[37]Hemberg S. Tanniferous kinds of Polygonum[J]. Leder,1951, (2):239-241.
[38]Li F, Zhan Z, Liu F, et al. Polyflavanostilbene A, a new flavanol-fused stilbene glycoside from Polygonum cuspidatum[J]. Organic Letters,2013,15(3):674-677.
[39]Zhang H, Zhang Q, Wang L, et al. Two new anthraquinone malonylglucosides from Polygonum cuspidatum[J]. Natural Product Research,2012,26(14):1323-1327.
[40]张彩宁.中药虎杖雌激素活性成分及其物质基础研究[D].中国科学院研究生院,2006.
[41]Kuznetsova Z P. Study of phenolic compounds from Japanese knotweed (Polygonum cuspidatum)[J]. Vestsi Akademii Navuk BSSR, Seryya Biyalagichnykh Navuk,1979:29-32.
[42]俸灵林.虎杖的化学成分及其质量研究[D].沈阳：沈阳药科大学,2003.
[43]Liu X, Wu L, Song G, et al. Study on chemical constituents of Polygonum cuspidatum Sieb[J]. Shenyang Yaoke Daxue Xuebao,1999,16(Suppl.):17-20.
[44]刘晓秋,于黎明,吴立军.虎杖化学成分研究(Ⅰ)[J].中国中药杂志,2003,28(1)：47-49.
[45]张海防,窦昌贵,刘晓华,等.虎杖提取物抗炎作用的实验研究[J].药学进展,2003,27(4)：230-233.
[46]Bralley E E, Greenspan P, Hargrove J L, et al. Topical anti-inflammatory activity of Polygonum cuspidatum extract in the TPA model of mouse ear inflammation[J]. Journal of Inflammation,2008,5(1):1.
[47]Banerjee S, Bueso-Ramos C, Aggarwal B B. Suppression of 7,12-Dimethylbenz (a) anthracene-induced mammary carcinogenesis in rats by resveratrol role of nuclear factor-κB, cyclooxygenase 2, and matrix metalloprotease 9[J]. Cancer Research,2002, 62(17):4945-4954.
[48]Manna S K, Mukhopadhyay A, Aggarwal B B. Resveratrol suppresses TNF-induced activation of nuclear transcription factors NF-κB, activator protein-1, and apoptosis:potential role of reactive oxygen intermediates and lipid peroxidation[J]. The Journal of Immunology,2000,164(12):6509-6519.
[49]Kumar A, Dhawan S, Aggarwal B B. Emodin (3-methyl-1,6, 8-trihydroxyanthraquinone) inhibits TNF-induced NF-kappaB activation, IkappaB degradation, and expression of cell surface adhesion proteins in human vascular endothelial cells.[J]. Oncogene,1998,17(7):913.
[50]Rhotc, He C, Sw L, et al. Inhibition of nitric oxide synthesis by coumarins from 264.7 cells Polygonum cuspidatum in LPS-activated RAW[J]. Saengyak Hakhoechi, 2001,32(3):181-188.
[51]Kimura Y, Okuda H, Kubo M. Effects of stilbenes isolated from medicinal plants on arachidonate metabolism and degranulation in human polymorphonuclear leukocytes[J]. Journal of Ethnopharmacology,1995,45(2):131-139.
[52]Shan B, Cai Y, Brooks J D, et al. Antibacterial properties of Polygonum cuspidatum roots and their major bioactive constituents[J]. Food Chemistry,2008,109(3): 530-537.
[53]Kukric Z Z, Topalic-Trivunovic L N. Antibacterial activity of cis-and trans-resveratrol isolated from Polygonum cuspidatum rhizome[J]. Acta Periodica Technologica 2006, (37):131-136.
[54]Song J, Yang T, Chang K, et al. In vitro effects of a fraction separated from Polygonum cuspidatum root on the viability, in suspension and biofilms, and biofilm formation of mutans streptococci [J]. Journal of Ethnopharmacology,2007,112(3): 419-425.
[55]K Y. Antimicrobial activity in natural resources. I. Antifungal and fungicidal activities of catechin and crude drugs against Trichophyton[J]. Toyama-ken Yakuji Kenkyusho Nenpo,1992,19:82-87.
[56]Lin H, Sun M, Wang Y, et al. Anti-HIV activities of the compounds isolated from Polygonum cuspidatum and Polygonum multiflorum[J]. Planta Medica,2010,76(09): 889-892.
[57]Y J, Hx W, Zy B, et al. Evaluation of antiviral effect of Polygonum cuspidatum water extract with a model of murine acquired immunodeficiency syndrome[J]. Virologica Sinica,1998,13(4):306-311.
[58]李凤新,孙振国,苏红,等.虎杖蒽醌提取物抗柯萨奇B3型病毒作用及机制研究[J].中草药,2008,39(11)：1699-1701.
[59]关文达,杨子峰,刘妮,等.白藜芦醇对多种呼吸道病毒作用体外实验研究[J].中药材,2008,31(9)：1388-1390.
[60]Chang J, Liu H, Wang K, et al. Ethanol extract of Polygonum cuspidatum inhibits hepatitis B virus in a stable HBV-producing cell line[J]. Antiviral Research,2005,66(1): 29-34.
[61]Park C, Lee Y, Kim J, et al. Inhibitory effects of Polygonum cuspidatum water extract (PCWE) and its component rasveratrol on acyl-coenzyme A-cholesterol acyltransferase activity for cholesteryl ester synthesis in HepG2 cells[J]. Vascular Pharmacology,2004,40(6):279-284.
[62]Xing W, Wu J, Jia M, et al. Effects of polydatin from Polygonum cuspidatum on lipid profile in hyperlipidemic rabbits[J]. Biomedicine & Pharmacotherapy,2009,63(7): 457-462.
[63]Du J, Sun L, Xing W, et al. Lipid-lowering effects of polydatin from Polygonum cuspidatum in hyperlipidemic hamsters[J]. Phytomedicine,2009,16(6):652-658.
[64]Sagar S M, Yance D, Wong R K. Natural health products that inhibit angiogenesis: a potential source for investigational new agents to treat cancer-Part 1[J]. Current Oncology,2006,13(1):14.
[65]Kimura Y, Okuda H. Resveratrol isolated from Polygonum cuspidatum root prevents tumor growth and metastasis to lung and tumor-induced neovascularization in Lewis lung carcinoma-bearing mice[J]. The Journal of Nutrition,2001,131(6): 1844-1849.
[66]Hsu C, Chan Y, Chang J. Antioxidant activity of extract from Polygonum cuspidatum[J]. Biological Research,2007,40(1):13-21.
[67]Pan Y, Zhang X, Wang H, et al. Antioxidant potential of ethanolic extract of Polygonum cuspidatum and application in peanut oil[J]. Food Chemistry,2007,105(4): 1518-1524.
[68]Zhang C Z, Wang S X, Zhang Y, et al. In vitro estrogenic activities of Chinese medicinal plants traditionally used for the management of menopausal symptoms[J]. Journal of Ethnopharmacology,2005,98(3):295-300.
[69]Zhang C, Wang X, Zhang X, et al. Bioassay-guided separation of citreorosein and other oestrogenic compounds From Polygonum cuspidatum[J]. Phytotherapy Research, 2009,23(5):740-741.
[70]Matsuda H, Shimoda H, Morikawa T, et al. Phytoestrogens from the roots of Polygonum cuspidatum (Polygonaceae):structure-requirement of hydroxyanthraquinones for estrogenic activity[J]. Bioorganic & Medicinal Chemistry Letters,2001,11(14): 1839-1842.
[71]Liu Z P, Li W X, Yu B, et al. Effects of trans-resveratrol from Polygonum cuspidatum on bone loss using the ovariectomized rat model [J]. Journal of Medicinal Food,2005,8(1):14-19.
[72]蔡宝昌,刘训红.常用中药材HPLC指纹图谱测定技术[M].北京：化学工业出版社,2005.2.
[73]易伦朝,吴海,梁逸曾.色谱指纹图谱与中药质量控制[J].色谱,2008,26(2)：166-171.
[74]谢培山.色谱指纹图谱分析是中草药质量控制的可行策略[J].中药新药与临床药理,2001,12(3)：141-151.
[75]彭静波,李强,雷海民,等.虎杖的功效成分组研究Ⅰ——延长凝血时间作用的谱效分析研究[J].中华中医药杂志,2011,26(8)：1737-1739.
[76]彭静波,李强,司银楚,等.虎杖的功效成分组研究Ⅱ——对全血黏度影响的谱效分析研究[J].中华中医药杂志,2011,26(9)：1964-1966.
[77]龙海林,李强,司银楚,等.虎杖的功效成分组研究Ⅲ——对血浆黏度影响的谱效分析研究[J].现代医药卫生,2012,28(020)：3080-3082.
[78]龙海林,雷海民,李强,等.虎杖的功效成分组研究Ⅳ-纤维蛋白原谱效分析的研究[J].中国医药指南,2012,10(33)：4-5.
[79]李强,夏晓晖,雷海民,等.虎杖功效成分组研究V-血沉沉降速率谱效关系研究[J].中医药学报,2011,39(5)：63.
[80]陈华国,赵宏宾,赵超,等.虎杖提取物HPLC指纹图谱归属分析及谱效关 系初探[J].中国药房,2010,(19)：1775-1776.
[81]罗志江,徐彦,吴建英,等.虎杖指纹图谱及其抗氧化活性的“谱-效”关系研究[J].西南大学学报(自然科学版),2012,34(1)：138-142.
[82]Zuo H, Shi Z, Hu X, et al. Prevalence of metabolic syndrome and factors associated with its components in Chinese adults[J]. Metabolism, Clinical and Experimental,2009,58(8):1102-1108.
[83]Alfaradhi M Z, Ozanne S E. Developmental programming in response to maternal overnutrition[J]. Frontiers in Genetics,2011,2,27.
[84]Despres J, Lemieux I, Bergeron J, et al. Abdominal obesity and the metabolic syndrome:contribution to global cardiometabolic risk[J]. Arteriosclerosis, Thrombosis, and Vascular Biology,2008,28(6):1039-1049.
[85]Dandona P, Aljada A, Bandyopadhyay A. Inflammation:the link between insulin resistance, obesity and diabetes[J]. Trends in Immunology,2004,25(1):4-7.
[86]Bradbury M W. Lipid metabolism and liver inflammation. I. Hepatic fatty acid uptake:possible role in steatosis[J]. American Journal of Physiology-Gastrointestinal and Liver Physiology,2006,290(2):G194-G198.
[87]Rector R S, Thyfault J P, Wei Y, et al. Non-alcoholic fatty liver disease and the metabolic syndrome:an update[J]. World Journal of Gastroenterology:WJG,2008,14(2): 185.
[88]Festi D, Colecchia A, Sacco T, et al. Hepatic steatosis in obese patients:clinical aspects and prognostic significance[J]. Obesity Reviews,2004,5(1):27-42.
[89]Unger R H. Lipid overload and overflow:metabolic trauma and the metabolic syndrome[J]. Trends in Endocrinology & Metabolism,2003,14(9):398-403.
[90]de Luca C, Olefsky J M. Stressed out about obesity and insulin resistance [J]. Nature Medicine,2006,12(1):41-42.
[91]Morino K, Petersen K F, Shulman G I. Molecular mechanisms of insulin resistance in humans and their potential links with mitochondrial dysfunction[J]. Diabetes,2006, 55(Suppl.2):S9-S15.
[92]Schinner S, Scherbaum W A, Bornstein S R, et al. Molecular mechanisms of insulin resistance[J]. Diabetic Medicine,2005,22(6):674-682.
[93]Steinberger J, Daniels S R. Obesity, Insulin Resistance, Diabetes, and Cardiovascular Risk in Children An American Heart Association Scientific Statement From the Atherosclerosis, Hypertension, and Obesity in the Young Committee (Council on Cardiovascular Disease in the Young) and the Diabetes Committee (Council on Nutrition, Physical Activity, and Metabolism)[J]. Circulation,2003,107(10):1448-1453.
[94]Cohen J C, Horton J D, Hobbs H H. Human fatty liver disease:old questions and new insights[J]. Science,2011,332(6037):1519-1523.
[95]Starley B Q, Calcagno C J, Harrison S A. Nonalcoholic fatty liver disease and hepatocellular carcinoma:a weighty connection[J]. Hepatology,2010,51(5):1820-1832.
[96]Ahmed M H, Byrne C D. Current treatment of non-alcoholic fatty liver disease[J]. Diabetes, Obesity and Metabolism,2009,11(3):188-195.
[97]Ioannides-Demos L L, Proietto J, Tonkin A M, et al. Safety of drug therapies used for weight loss and treatment of obesity[J]. Drug Safety,2006,29(4):277-302.
[98]Vermaak I, Viljoen A M, Hamman J H. Natural products in anti-obesity therapy[J]. Natural Product Reports,2011,28(9):1493-1533.
[99]王辉,杨再刚,燕树勋,等.虎杖总蒽醌对糖尿病肾病早期血瘀模型大鼠脂代谢及血液流变性的影响[J].中国实验方剂学杂志,2010,16(16).
[100]郑晓媛.虎杖提取物对α-葡萄糖苷酶的抑制作用研究[D].四川大学,2007.
[101]沈忠明,殷建伟,袁海波.虎杖鞣质的降血糖作用研究[J].天然产物研究与开发,2004,16(3).
[102]Ricchi M, Odoardi M R, Carulli L, et al. Differential effect of oleic and palmitic acid on lipid accumulation and apoptosis in cultured hepatocytes[J]. Journal of Gastroenterology and Hepatology,2009,24(5):830-840.
[103]Morikawa T, Ninomiya K, Zhang Y, et al. Flavonol glycosides with lipid accumulation inhibitory activity from Sedum sarmentosum[J].Phytochemistry Letters, 2012,5(1):53-58.
[104]方飞,罗明俐,苏楠,等.桑叶提取物对胰岛素抵抗HepG2细胞葡萄糖摄取的影响及其分子机制[J].药学学报,2012,47(011)：1452-1456.
[105]杨桂枝,高小平,晏菊芳,等.GOD-POD法微量化测定方法的建立及其在3T3-L1脂肪细胞和HepG2细胞糖摄取中的应用[J].四川解剖学杂志,2003：12-15.
[106]姜斐,姚楠,钱士辉,等.女贞子中8个化合物对胰岛素抵抗HepG2细胞葡萄糖消耗的影响[J].海峡药学,2010,(001)：164-167.
[107]Huang Y, Lin C, Huang S, et al. Effect and mechanism of ginsenosides CK. and Rgl on stimulation of glucose uptake in 3T3-L1 adipocytes[J]. Journal of Agricultural and Food Chemistry,2010,58(10):6039-6047.
[108]Ann Y M, Kim S K, Kang J S, et al. Platycodon grandiflorum modifies adipokines and the glucose uptake in high-fat diet in mice and L6 muscle cells[J]. Journal of Pharmacy and Pharmacology,2012,64(5):697-704.
[109]Noh J, Lee I, Ly S, et al. A Phellinus baumii extract reduces obesity in high-fat diet-fed mice and absorption of triglyceride in lipid-loaded mice[J]. Journal of Medicinal Food,2011,14(3):209-218.
[110]Zou C, Wang Y, Shen Z.2-NBDG as a fluorescent indicator for direct glucose uptake measurement[J]. Journal of Biochemical and Biophysical Methods,2005,64(3): 207-215.
[111]李晨钟,张素华,舒昌达,等.胰岛素抵抗细胞模型的胰岛素降解酶活性与胰岛素敏感性的关系[J].第一军医大学学报,2002,22(2)：151-154.
[112]李兰芳,黄秀清,原慧萍,等.吡格列酮改善胰岛素抵抗状态下氧化应激水平及分子机制探讨[J].中国临床药理学与治疗学,2009,14(3)：245-249.
[113]李美妮,韩蕊莲,韩建萍,等.大黄与易混伪品土大黄,虎杖原植物的ITS2序列鉴定[J].环球中医药,2012,3：10.
[114]Frederich M, Wauters J N, Tits M, et al. Quality Assessment of Polygonum cuspidatum and Polygonum multiflorum by 1H NMR metabolite fingerprinting and profiling analysis[J]. Planta Medica,2011,77:81-86.
[115]Qian G, Leung S, Lu G, et al. Differentiation of Rhizoma et Radix Polygoni cuspidati from closely related herbs by HPLC fingerprinting[J]. Chemical and Pharmaceutical Bulletin,2006,54(8):1179-1186.
[116]Hebert P D, Cywinska A, Ball S L. Biological identifications through DNA barcodes[J]. Proceedings of the Royal Society of London. Series B:Biological Sciences, 2003,270(1512):313-321.
[117]Gregory T R. DNA barcoding does not compete with taxonomy[J]. Nature,2005, 434(7037):1067.
[118]Miller S E. DNA barcoding and the renaissance of taxonomy [J]. Proceedings of the National Academy of Sciences,2007,104(12):4775-4776.
[119]陈士林,宋经元,姚辉.药用植物DNA条形码鉴定策略及关键技术分析[J].中国天然药物,2009,7(5)：322-327.
[120]陈士林,姚辉,宋经元,等.基于DNA barcoding(条形码)技术的中药材鉴定[J].世界科学技术：中医药现代化,2007,9(3)：7-12.
[121]罗焜,陈科力,刘义梅,等.高良姜及其混淆品的分子鉴定[J].世界科学技术：中医药现代化,2011,13(2)：400-406.
[122]罗焜,陈士林,陈科力,等.基于芸香科的植物通用DNA条形码研究[J].中国科学：生命科学,2010,(4)：342-351.
[123]Pang X, Song J, Zhu Y, et al. Applying plant DNA barcodes for Rosaceae species identification[J]. Cladistics,2011,27(2):165-170.
[124]Gao T, Yao H, Song J, et al. Identification of medicinal plants in the family Fabaceae using a potential DNA barcode ITS2[J]. Journal of Ethnopharmacology,2010, 130(1):116-121.
[125]朱英杰,陈士林,姚辉,等.重楼属药用植物DNA条形码鉴定研究[J].药学学报,2010,45(3)：376-382.
[126]Yao H, Song J, Liu C, et al. Use of ITS2 region as the universal DNA barcode for plants and animals[J]. PloS One,2010,5(10):e13102.
[127]Kress W J, Erickson D L. A two-locus global DNA barcode for land plants:the coding rbcL gene complements the non-coding trnH-psbA spacer region[J], PLoS One, 2007,2(6):e508.
[128]Lahaye R, van der Bank M, Bogarin D, et al. DNA barcoding the floras of biodiversity hotspots[J]. Proceedings of the National Academy of Sciences,2008,105(8): 2923-2928.
[129]Chase M W, Cowan R S, Hollingsworth P M, et al. A proposal for a standardised protocol to barcode all land plants[J]. Taxon,2007:295-299.
[130]Chen S, Yao H, Han J, et al. Validation of the ITS2 region as a novel DNA barcode for identifying medicinal plant species[J]. PloS one,2010,5(1):e8613.
[131]Hollingsworth P M, Forrest L L, Spouge J L, et al. A DNA barcode for land plants[J]. Proceedings of the National Academy of Sciences,2009,106(31): 12794-12797.
[132]Kress W J, Wurdack K J, Zimmer E A, et al. Use of DNA barcodes to identify flowering plants[J]. Proceedings of the National Academy of Sciences of the United States of America,2005,102(23):8369-8374.
[133]Chase M W, Salamin N, Wilkinson M, et al. Land plants and DNA barcodes: short-term and long-term goals[J]. Philosophical Transactions of the Royal Society B: Biological Sciences,2005,360(1462):1889-1895.
[134]Gonzalez M A, Baraloto C, Engel J, et al. Identification of Amazonian trees with DNA barcodes[J]. PLoS One,2009,4(10):e7483.
[135]E P. Wanted:a barcode for plants[J]. Science,2007,318:190-191.
[136]Keller A, Schleicher T, Schultz J, et al.5.8 S-28S rRNA interaction and HMM-based ITS2 annotation.[J]. Gene,2009,430(1-2):50.
[137]Muller T, Philippi N, Dandekar T, et al. Distinguishing species[J]. RNA,2007, 13(9):1469-1472.
[138]Miao M, Warren A, Song W, et al. Analysis of the internal transcribed spacer 2 (ITS2) region of scuticociliates and related taxa (Ciliophora, Oligohymenophorea) to infer their evolution and phylogeny[J]. Protist,2008,159(4):519-533.
[139]Shaw J, Lickey E B, Beck J T, et al. The tortoise and the hare Ⅱ:relative utility of 21 noncoding chloroplast DNA sequences for phylogenetic analysis[J]. American Journal of Botany,2005,92(1):142-166.
[140]Yao H, Song J, Ma X, et al. Identification of Dendrobium species by a candidate DNA barcode sequence:the chloroplast psbA-trnH intergenic region[J]. Planta Medica, 2009,75(06):667-669.
[141]Zhao Z, Xiao P. Encyclopedia of medicinal plants (Volume II)[M]. Shanghai: World Publishing Corporation,2007.235-239.
[142]马云桐,万德光,宋良科.虎杖药材HPLC指纹图谱研究[J].中国中药杂志,2006,31(12)：972-974.
[143]雷海民,毕葳,李强,等.虎杖药材指纹图谱研究[J].中草药,2006,37(7)：1084-1086.
[144]张明昶,彭小冰,苗旭辉,等.中药虎杖特征指纹图谱研究[J].中国实验方剂学杂志,2011,17(1)：53.
[145]李宇翔,王丽,唐书明,等.虎杖HPLC指纹图谱研究与应用[J].中国实验方剂学杂志,2010,16(5)：78.
[146]马培,许利嘉,刘延泽,等.虎杖药材UPLC特征图谱研究[J].中国实验方剂学杂志,2012,8(2).
[147]Ma P, Luo K, Peng Y, et al. Quality control of Polygonum cuspidatum by UPLC-PDA and related statistical analysis[J]. Journal of Liquid Chromatography & Related Technologies, doi:10.1080/10826076.2012.723096.
[148]王玺,王文宇,张克荣,等.中药HPLC指纹图谱相似性研究的探讨[J].沈阳药科大学学报,2003,20(5)：360-362.
[149]孟庆华,刘永锁,蒋淑敏,等.色谱指纹图谱综合信息指数在中药质量控制中的应用研究[J].中国天然药物,2004,2(6)：359-364.
[150]Xie G X, Ni Y, Su M M, et al. Application of ultra-performance LC-TOF MS metabolite profiling techniques to the analysis of medicinal Panax herbs[J]. Metabolomics,2008,4(3):248-260.
[151]云南省食品药品监督管理局.云南省中药材标准·第一册[M].昆明：云南美术 出版社,2005.33.
[152]Chandrasekar D, Madhusudhana K, Ramakrishna S, et al. Determination of DPPH free radical scavenging activity by reversed-phase HPLC:A sensitive screening method for polyherbal formulations[J]. Journal of Pharmaceutical and Biomedical Analysis,2006, 40(2):460-464.
[153]Qiu J, Chen L, Zhu Q, et al. Screening natural antioxidants in peanut shell using DPPH-HPLC-DAD-TOF/MS methods[J]. Food Chemistry,2012,135(4):2366-2371.
[154]Tang D, Li H J, Chen J, et al. Rapid and simple method for screening of natural antioxidants from Chinese herb Flos Lonicerae Japonicae by DPPH-HPLC-DAD-TOF/MS[J]. Journal of Separation Science,2008,31(20): 3519-3526.
[155]Shui G, Leong L P. Screening and identification of antioxidants in biological samples using high-performance liquid chromatography-mass spectrometry and its application on Salacca edulis Reinw[J]. Journal of Agricultural and Food Chemistry, 2005,53(4):880-886.
[156]Rice-Evans C A, Miller N J, Bolwell P G, et al. The relative antioxidant activities of plant-derived polyphenolic flavonoids[J]. Free Radical Research,1995,22(4): 375-383.
[157]Vinson J A, Dabbagh Y A, Serry M M, et al. Plant flavonoids, especially tea flavonols, are powerful antioxidants using an in vitro oxidation model for heart disease[J]. Journal of Agricultural and Food Chemistry,1995,43(11):2800-2802.
[158]Laguerre M, Lecomte J, Villeneuve P. Evaluation of the ability of antioxidants to counteract lipid oxidation:existing methods, new trends and challenges[J]. Progress in Lipid Research,2007,46(5):244-282.