海带（Laminaria japonica Aresch）多酚的提取、分离及其抗肿瘤、抗菌活性研究
|
摘要
植物多酚与人类的生活息息相关,作为一类具有多种生物活性的天然物质,它对人的消化、营养、健康产生影响,已在食品、药品、化妆品等工业中得到了广泛的应用。本论文以鲜海带(Laminaria japonica Aresch)为原料,系统地研究了海带多酚(KP)的制备、分离纯化、抗肿瘤、抑菌活性,并初步探讨了其理化性质和作用机理,为深入研究海藻多酚提供了基础数据,同时也为海带的高值化利用及新的抗肿瘤药物、食品保鲜剂的开发提供一种新的思路。
本论文的研究内容包括以下6个部分:
1.海带多酚提取率测定方法的选择:
海带可以综合利用提取多酚和其它副产物。通过酒石酸铁比色法和Folin-Denis法两种多酚定量分析方法的比较,表明以没食子酸丙酯为参照物的酒石酸铁比色法方便、快速、可靠,可以作为本次实验定量分析手段。另外,在同样的提取条件下,鲜海带总酚含量要比干海带的总酚含量略高。
2.海带多酚提取工艺条件的优化:
采用鲜海带为原料,以超声波、微波复合浸提为前处理手段,以多酚的提取率为指标,在单因素实验的基础上采用正交实验对KP浸提工艺参数进行了优化,结果表明:采用料液比1:7(g/mL),乙醇浓度85%,浸提温度70℃,浸提次数2次,浸提时间4h,提取效果最佳,KP提取率为2.08%。
3.海带多酚粗提物抗肿瘤、抗菌活性的研究:
以MTT法分别测定了KP粗提物对人肺腺癌A549细胞、鼠白血病P388细胞、人肝癌BEL-7402细胞和人宫颈癌Hela细胞的抗肿瘤活性;以平板打孔抑制法分别测定了KP粗提物对金黄色葡萄球菌、枯草芽孢杆菌、大肠杆菌、绿脓杆菌、痢疾志贺菌、产气肠杆菌、普通变形杆菌、白色念珠菌、青霉、灰霉、黄曲霉的抑菌活性。结果表明:KP粗提物具有一定的抗肿瘤活性,在100μg/mL时,对A549、P388、BEL-7402、Hela细胞的抑制率分别为27.40±4.71%,43.44±1.86%,30.20±1.16%,29.68±2.61%。镜检发现细胞形态都发生不同程度的变化。肿瘤细胞的生长抑制活性量效关系表明KP对P388的IC50为120μg/mL,对BEL-7402的IC50大于200μg/mL,细胞毒作用呈剂量和时间依赖性,对P388的抑制活性要高于对BEL-7402的抑制活性。抑菌实验也显示KP具有广谱抗微生物性能,对供试的2种革兰氏阳性菌和5种革兰氏阴性菌都有一定程度的抑制活性,量效关系均呈剂量依赖型,最小抑菌浓度(MIC)为10μg/mL～40μg/mL。另外,多酚对革兰氏阴性细菌中的大肠杆菌、绿脓杆菌抑制效果较明显,对真菌如青霉、白色念珠菌亦有抑制作用。
4.海带多酚的分离纯化及其各部分活性检测:
利用有机溶剂萃取分离、AB-8大孔树脂吸附分离、SephadexLH-20凝胶层析等技术对KP进行分离纯化,最后得到4个峰组分,分别为A1、A2、B1、B2,并筛选出了具有较高活性的多酚组分。各组分肿瘤抑制活性大小依次为:A2 > A1 > B2 > B1,浓度为70.42μg/mL时,组分A2对BEL-7402和P388的抑制率分别为61.96±7.02%,40.47±8.70%;各组分的抑菌活性大小依次为:A2 > B2 > A1 > B1,在酚浓度为70.42μg/mL、45.68μg/mL的条件下,对4种菌均有抑制效果且比粗提物和树脂分离产物同等浓度下抑菌圈直径大。
5.海带多酚理化性质初步研究:
对KP的物理化学性质如溶解性、折光率、特征显色反应、pH稳定性、热稳定性及光谱特征进行了初步研究。结果表明:KP易溶于中极性溶剂,而在非极性溶剂和极性溶剂中溶解性较差;在不同溶剂中的折光率大小依次为乙酸乙酯>乙醇>丙酮>甲醇>水>氯仿=乙醚;特征显色反应表明KP具有酚类物质的共性。另外,KP在pH2.0～11.0和40～100℃的范围内都有肿瘤抑制活性,具有良好的pH稳定性和热稳定性。组分A2与PG、间苯三酚的紫外吸收谱图具有相似之处。组分A1、A2、B2与PG、连苯三酚的红外谱图都具有明显的酚羟基伸缩振动;都有芳香环骨架伸缩振动;在指纹区都有酚的特征吸收峰,在官能团区也较为相似,初步推断KP具有PG或连苯三酚相近或相似的结构。
6.海带多酚的抗肿瘤活性和抗菌活性机理初步探讨:
以显微镜观察药物处理前后细胞的形态变化,考察了多酚对自由基、致癌物的清除能力;以流式细胞术分析了多酚对肿瘤细胞的凋亡率及对细胞增殖周期的影响,并测定了菌体系统的紫外吸收变化。结果表明:KP处理P388和BEL-7402细胞48h后,镜检细胞形态发生较大变化,与正常对照组差异显著;在100μg/mL的浓度下,KP具有一定的清除自由基和致癌物的活性,对·OH、O2·-、NO2-清除率分别为75.37±5.68 %、55.48±4.35%、50.51±6.46%。流式细胞术表明多酚对P388肿瘤细胞的凋亡率及细胞增殖周期存在影响,其中A2组分凋亡峰极为显著。加入多酚的菌悬液280nm处的吸光度均明显增高,且对革兰氏阴性菌更为明显。
As a class of natural material, which is rich in various biological activities, plant polyphenol links closely with human life. It has effected on human digestion, nutrition, health and has been widespreadly used in the area of food, drug and cosmetic industry.
In this paper, the extraction, isolation, anti-tumor activity and anti-bacterial activity of the polyphenols from fresh Laminaria japonica Aresch were studied systematically, and some characteristics, mechanism and application were also investigated. This work may provide primary data for further researching on the relationship between the structure and function of phlorotannins, as well as provide a new way for high-value utilization of kelp and exploiting some new anti-tumor medicine and food antiseptic.
This study includes six parts of content as follows:
1. Selection of measuement method of phlorotannins on exudation rate The kelp could be synthetically utilized to extract phlorotannins and other by-products. Based on comparison study on the two methods of determination phlorotannins, ferrous tartrate colorimetry and Folin-Denis method, ferrous tartrate colorimetry with PG as reference material was chosen because of its facility and credibility. Phlorotannins content of fresh kelp was slightly higher than dry kelp under the same extraction conditions.
2. Determination of the optimal conditions for extraction of phlorotannins
Adopt fresh kelp as materials, ultrasonic and microwave extraction as pretreatment, extraction rate of phlorotannins as target, on the base of the single factor experiment and orthogonal test, the optimal extraction conditions of kelp polyphenols were researched. The results show that optimal extraction conditions of kelp polyphenols from Laminaria japonica Aresch are the ratio of sample weight to the solvent volume 1:7(g/mL), 85% ethanol, temperature 70℃, extraction times 2, extraction time 4h, and the exudation rate of kelp polyphenols is 2.08%.
3. Investigation on anti-tumor activity and anti-bacterial activity of the crude extracts of brown algae phlorotannins
The anti-tumor activity of crude extract of phlorotannins for human lung adenocarcinoma cell (A549), murine leukemic cell (P388), human hepatocellular carcinoma cell (BEL-7402) and human cervical cancer cell (Hela) had been determined respectively by MTT method in vitro. The results showed crude extracts has anti-tumor activity and the inhibitory rate were 27.40±4.71%, 43.44±1.86%, 30.20±1.16%, 29.68±2.61%, respectively at the concentration of 100μg/mL. The result of the microscope detection showed that the morphologic features of tumor cells were changed from the normal control group. Dosage-effect relationship showed the half-maximal inhibitory concentration (IC50) for P388 cell and BEL-7402 cell were 120μg/mL and >200μg/mL respectively. Moreover, the cytotoxic effect to P388 and BEL-7402 seems to be dose-dependent and time-dependent and the inhibitory rate for P388 was superior to that of BEL-7402.
The anti-bacterial activity of crude extracts of phlorotannins had been determined by flat plate slotting to inhibit growth method in vitro, respectively. The results of bacteriostatic experiment showed that 2 species of gram-positive cocci and 5 species of gram-negative bacilli were inhibited by crude extract which had wide antimicrobial performance. Dosage-effect relationship was dose-dependent and the minimal inhibitory concentration (MIC) was 10μg/mL～40μg/mL. Meanwhile, the inhibitory effect of crude extract to Escherichia coli, Pseudomonsa aeruginosa was relatively obvious and fungi such as Penicillium, Candida albicans were also inhibited.
4. Isolation and purification process of the extract crudes
The crude extracts could be divided into 4 kinds of bioactive fractions (A1, A2, B1, B2) after extraction with organic solvent, adsorption and desorption with AB-8 macroporous resin, column chromatography with sephadexLH-20. All bioactive fractions were compared in each purification process. Screening results of antitumor and antibacterial activity showed A2 > A1> B2 > B1 and A2 > B2 > A1 > B1, respectively. And the inhibitory rate of fraction A2 was 61.96±7.02% and 40.47±8.70% to BEL-7402 and P388 respectively at the concentration of 70.42μg/mL, and antibacterial activity was also increased compared to front parts.
5. Primary research on the physical and chemical characteristics of phlorotannins
The physical and chemic characteristics of phlorotannins such as dissolubility, light refraction, characteristic color reaction, pH stability, heat stability and spectral feature were determined primaryly. The results showed that the dissolubility of phlorotannins in CH3OH, CH3CH2OH, C2H5COOC2H5 were better than that in H2O, CH3Cl, C2H5OC2H5. Refractive index of phlorotannins in different solvents was C2H5COOC2H5 > CH3CH2OH > CH3COCH3 > CH3OH > H2O > CH3Cl = C2H5OC2H5. Characteristic color reaction showed kelp phlorotannins had the commonness of phenols. Otherwise, kelp phlorotannins had pH stability, thermal stability between pH 2.0～11.0 and 40℃～100℃. Ultraviolet absorption spectra of component A2 was similar to PG and phloroglucinol dehydrate. IR spectras of fraction A1, fraction A2 fraction B2, PG and pyrogallol had obvious expansion and vibration of phenol hydroxyl, aromatic rings. Kelp phlorotannins compound is preliminarily deduced as similar structure of PG or pyrogallol.
6. Primary discussion of anti-tumor activity and anti-bacterial activity mechanism of phlorotannins
The change of cell morphology was observed by fluorescent microscopy, and scavenging ability of free radicals and carcinogenic agents were discussed. At the same time, we analysed cell apoptosis, cell cycle by flow cytometry and detected UV absorption. The results showed that microscope detection the morphologic features of P388 and BEL-7402 tumor cells were changed from the normal control group after treated 48h. The scavenging rate of·OH, O2·-, NO2–were 75.37±5.68 %, 55.48±4.35%, 50.51±6.46%, respectively. The anti-tumor activity of phlorotannins was related with the cell apoptosis and cell proliferation cycle, especially obvious of A2 apoptosis peak, but the molecular mechanisms remain unclear. Ultraviolet absorption of the system containing phlorotannins and bacteria was increased at 280nm, especially for G-.
本论文的研究内容包括以下6个部分:
1.海带多酚提取率测定方法的选择:
海带可以综合利用提取多酚和其它副产物。通过酒石酸铁比色法和Folin-Denis法两种多酚定量分析方法的比较,表明以没食子酸丙酯为参照物的酒石酸铁比色法方便、快速、可靠,可以作为本次实验定量分析手段。另外,在同样的提取条件下,鲜海带总酚含量要比干海带的总酚含量略高。
2.海带多酚提取工艺条件的优化:
采用鲜海带为原料,以超声波、微波复合浸提为前处理手段,以多酚的提取率为指标,在单因素实验的基础上采用正交实验对KP浸提工艺参数进行了优化,结果表明:采用料液比1:7(g/mL),乙醇浓度85%,浸提温度70℃,浸提次数2次,浸提时间4h,提取效果最佳,KP提取率为2.08%。
3.海带多酚粗提物抗肿瘤、抗菌活性的研究:
以MTT法分别测定了KP粗提物对人肺腺癌A549细胞、鼠白血病P388细胞、人肝癌BEL-7402细胞和人宫颈癌Hela细胞的抗肿瘤活性;以平板打孔抑制法分别测定了KP粗提物对金黄色葡萄球菌、枯草芽孢杆菌、大肠杆菌、绿脓杆菌、痢疾志贺菌、产气肠杆菌、普通变形杆菌、白色念珠菌、青霉、灰霉、黄曲霉的抑菌活性。结果表明:KP粗提物具有一定的抗肿瘤活性,在100μg/mL时,对A549、P388、BEL-7402、Hela细胞的抑制率分别为27.40±4.71%,43.44±1.86%,30.20±1.16%,29.68±2.61%。镜检发现细胞形态都发生不同程度的变化。肿瘤细胞的生长抑制活性量效关系表明KP对P388的IC50为120μg/mL,对BEL-7402的IC50大于200μg/mL,细胞毒作用呈剂量和时间依赖性,对P388的抑制活性要高于对BEL-7402的抑制活性。抑菌实验也显示KP具有广谱抗微生物性能,对供试的2种革兰氏阳性菌和5种革兰氏阴性菌都有一定程度的抑制活性,量效关系均呈剂量依赖型,最小抑菌浓度(MIC)为10μg/mL～40μg/mL。另外,多酚对革兰氏阴性细菌中的大肠杆菌、绿脓杆菌抑制效果较明显,对真菌如青霉、白色念珠菌亦有抑制作用。
4.海带多酚的分离纯化及其各部分活性检测:
利用有机溶剂萃取分离、AB-8大孔树脂吸附分离、SephadexLH-20凝胶层析等技术对KP进行分离纯化,最后得到4个峰组分,分别为A1、A2、B1、B2,并筛选出了具有较高活性的多酚组分。各组分肿瘤抑制活性大小依次为:A2 > A1 > B2 > B1,浓度为70.42μg/mL时,组分A2对BEL-7402和P388的抑制率分别为61.96±7.02%,40.47±8.70%;各组分的抑菌活性大小依次为:A2 > B2 > A1 > B1,在酚浓度为70.42μg/mL、45.68μg/mL的条件下,对4种菌均有抑制效果且比粗提物和树脂分离产物同等浓度下抑菌圈直径大。
5.海带多酚理化性质初步研究:
对KP的物理化学性质如溶解性、折光率、特征显色反应、pH稳定性、热稳定性及光谱特征进行了初步研究。结果表明:KP易溶于中极性溶剂,而在非极性溶剂和极性溶剂中溶解性较差;在不同溶剂中的折光率大小依次为乙酸乙酯>乙醇>丙酮>甲醇>水>氯仿=乙醚;特征显色反应表明KP具有酚类物质的共性。另外,KP在pH2.0～11.0和40～100℃的范围内都有肿瘤抑制活性,具有良好的pH稳定性和热稳定性。组分A2与PG、间苯三酚的紫外吸收谱图具有相似之处。组分A1、A2、B2与PG、连苯三酚的红外谱图都具有明显的酚羟基伸缩振动;都有芳香环骨架伸缩振动;在指纹区都有酚的特征吸收峰,在官能团区也较为相似,初步推断KP具有PG或连苯三酚相近或相似的结构。
6.海带多酚的抗肿瘤活性和抗菌活性机理初步探讨:
以显微镜观察药物处理前后细胞的形态变化,考察了多酚对自由基、致癌物的清除能力;以流式细胞术分析了多酚对肿瘤细胞的凋亡率及对细胞增殖周期的影响,并测定了菌体系统的紫外吸收变化。结果表明:KP处理P388和BEL-7402细胞48h后,镜检细胞形态发生较大变化,与正常对照组差异显著;在100μg/mL的浓度下,KP具有一定的清除自由基和致癌物的活性,对·OH、O2·-、NO2-清除率分别为75.37±5.68 %、55.48±4.35%、50.51±6.46%。流式细胞术表明多酚对P388肿瘤细胞的凋亡率及细胞增殖周期存在影响,其中A2组分凋亡峰极为显著。加入多酚的菌悬液280nm处的吸光度均明显增高,且对革兰氏阴性菌更为明显。
As a class of natural material, which is rich in various biological activities, plant polyphenol links closely with human life. It has effected on human digestion, nutrition, health and has been widespreadly used in the area of food, drug and cosmetic industry.
In this paper, the extraction, isolation, anti-tumor activity and anti-bacterial activity of the polyphenols from fresh Laminaria japonica Aresch were studied systematically, and some characteristics, mechanism and application were also investigated. This work may provide primary data for further researching on the relationship between the structure and function of phlorotannins, as well as provide a new way for high-value utilization of kelp and exploiting some new anti-tumor medicine and food antiseptic.
This study includes six parts of content as follows:
1. Selection of measuement method of phlorotannins on exudation rate The kelp could be synthetically utilized to extract phlorotannins and other by-products. Based on comparison study on the two methods of determination phlorotannins, ferrous tartrate colorimetry and Folin-Denis method, ferrous tartrate colorimetry with PG as reference material was chosen because of its facility and credibility. Phlorotannins content of fresh kelp was slightly higher than dry kelp under the same extraction conditions.
2. Determination of the optimal conditions for extraction of phlorotannins
Adopt fresh kelp as materials, ultrasonic and microwave extraction as pretreatment, extraction rate of phlorotannins as target, on the base of the single factor experiment and orthogonal test, the optimal extraction conditions of kelp polyphenols were researched. The results show that optimal extraction conditions of kelp polyphenols from Laminaria japonica Aresch are the ratio of sample weight to the solvent volume 1:7(g/mL), 85% ethanol, temperature 70℃, extraction times 2, extraction time 4h, and the exudation rate of kelp polyphenols is 2.08%.
3. Investigation on anti-tumor activity and anti-bacterial activity of the crude extracts of brown algae phlorotannins
The anti-tumor activity of crude extract of phlorotannins for human lung adenocarcinoma cell (A549), murine leukemic cell (P388), human hepatocellular carcinoma cell (BEL-7402) and human cervical cancer cell (Hela) had been determined respectively by MTT method in vitro. The results showed crude extracts has anti-tumor activity and the inhibitory rate were 27.40±4.71%, 43.44±1.86%, 30.20±1.16%, 29.68±2.61%, respectively at the concentration of 100μg/mL. The result of the microscope detection showed that the morphologic features of tumor cells were changed from the normal control group. Dosage-effect relationship showed the half-maximal inhibitory concentration (IC50) for P388 cell and BEL-7402 cell were 120μg/mL and >200μg/mL respectively. Moreover, the cytotoxic effect to P388 and BEL-7402 seems to be dose-dependent and time-dependent and the inhibitory rate for P388 was superior to that of BEL-7402.
The anti-bacterial activity of crude extracts of phlorotannins had been determined by flat plate slotting to inhibit growth method in vitro, respectively. The results of bacteriostatic experiment showed that 2 species of gram-positive cocci and 5 species of gram-negative bacilli were inhibited by crude extract which had wide antimicrobial performance. Dosage-effect relationship was dose-dependent and the minimal inhibitory concentration (MIC) was 10μg/mL～40μg/mL. Meanwhile, the inhibitory effect of crude extract to Escherichia coli, Pseudomonsa aeruginosa was relatively obvious and fungi such as Penicillium, Candida albicans were also inhibited.
4. Isolation and purification process of the extract crudes
The crude extracts could be divided into 4 kinds of bioactive fractions (A1, A2, B1, B2) after extraction with organic solvent, adsorption and desorption with AB-8 macroporous resin, column chromatography with sephadexLH-20. All bioactive fractions were compared in each purification process. Screening results of antitumor and antibacterial activity showed A2 > A1> B2 > B1 and A2 > B2 > A1 > B1, respectively. And the inhibitory rate of fraction A2 was 61.96±7.02% and 40.47±8.70% to BEL-7402 and P388 respectively at the concentration of 70.42μg/mL, and antibacterial activity was also increased compared to front parts.
5. Primary research on the physical and chemical characteristics of phlorotannins
The physical and chemic characteristics of phlorotannins such as dissolubility, light refraction, characteristic color reaction, pH stability, heat stability and spectral feature were determined primaryly. The results showed that the dissolubility of phlorotannins in CH3OH, CH3CH2OH, C2H5COOC2H5 were better than that in H2O, CH3Cl, C2H5OC2H5. Refractive index of phlorotannins in different solvents was C2H5COOC2H5 > CH3CH2OH > CH3COCH3 > CH3OH > H2O > CH3Cl = C2H5OC2H5. Characteristic color reaction showed kelp phlorotannins had the commonness of phenols. Otherwise, kelp phlorotannins had pH stability, thermal stability between pH 2.0～11.0 and 40℃～100℃. Ultraviolet absorption spectra of component A2 was similar to PG and phloroglucinol dehydrate. IR spectras of fraction A1, fraction A2 fraction B2, PG and pyrogallol had obvious expansion and vibration of phenol hydroxyl, aromatic rings. Kelp phlorotannins compound is preliminarily deduced as similar structure of PG or pyrogallol.
6. Primary discussion of anti-tumor activity and anti-bacterial activity mechanism of phlorotannins
The change of cell morphology was observed by fluorescent microscopy, and scavenging ability of free radicals and carcinogenic agents were discussed. At the same time, we analysed cell apoptosis, cell cycle by flow cytometry and detected UV absorption. The results showed that microscope detection the morphologic features of P388 and BEL-7402 tumor cells were changed from the normal control group after treated 48h. The scavenging rate of·OH, O2·-, NO2–were 75.37±5.68 %, 55.48±4.35%, 50.51±6.46%, respectively. The anti-tumor activity of phlorotannins was related with the cell apoptosis and cell proliferation cycle, especially obvious of A2 apoptosis peak, but the molecular mechanisms remain unclear. Ultraviolet absorption of the system containing phlorotannins and bacteria was increased at 280nm, especially for G-.
引文
[1] Paul VJ,Littler MM,Littler DS,et al.Evidence for chemical defence in tropical green alga Caulerpa ashmeadii (Caulerpaceae: Chlorophyta): isolation of new bioactive sesquiterpenoids [J].J Chem Ecol,1987,13(5):1171～1185.
[2] Handley JT,Blackman AJ.Secondary metabolites from the marine alga Caulerpa brownii (Chlorophyta)[J].Australian Journal of Chemistry,2005,58(1):39～46.
[3] Erickson AA,Paul VJ,Van Alstyne KL,Kwiatkowski LM.Palatability of macroalgae that use different types of chemical defenses[J].Journal of Chemical Ecology,2006,31(9):1883～1895.
[4]纪明侯.海藻化学[M].北京:科学出版社,1997:628.
[5]徐秀丽,范晓,宋福行,等.褐藻小粘膜藻中的溴酚化合物[J].中国中药杂志,2005,30(1):39～41.
[6]严小军.中国常见褐藻的多酚含量测定[J].海洋科学集刊,1996,37:61～64.
[7] Kanget.Study on the extraction condition and quantity of polyphenols in a dry brown algae Ecklonia cava[J].Bull Mar Res,1990,14:25～38.
[8]Barwell CJ,Blunden G,Manandhar PD.Isolation and characterization of brown algal polyphenols as inhibitors ofα-amylase,lipase,and trypsin [J].J Appl Phycol,1989,1:319-323.
[9] Keusgen M,Glombitza KW.Phlorethols,fuhalols,and their derivatives from the brown alga Sargassum spinulligerum[J].Phytochemistry,1995,38(4):975～985.
[10]Glombitza KW,Keusgen M,Hauperich S.Fucophlorethols from the algae Sargassum spinuligerum and Cystophora torulosa[J].Phytochemistry,1997,46(8):1417～1422.
[11] Sailler B,Glombitza KW.Phlorethols and fucophlorethols from the brown alga Cystophora retroflexa[J].Phytochemistry,1999,50:869～881.
[12]Glombitza KW,Schmidt A.Trihydroxyphlorethols from the brown alga Carpophyllum angustifolium[J].Phytochemistry,1999,51:1095～1100.
[13] Smller B,Glombitza KW.Halogenated phlorethols and fucophlorethols from the brown alga Cystophora retroflexa[J]. Nat Toxins,1999,7(2):57～62.
[14] Glombitza KW,Schmidt A.Nonhalogenated and halogenated phlorotannins from the brown alga Carpophyllum angustifolium[J].J Nat Prod,1999,62(9):1238～1240.
[15]陈予敏,严小军,范晓.褐藻多酚的结构单元及生成机理[J].海洋与湖沼,1997,28(3):225～231.
[16]郭孝武.超声提取与常规提取对部分中药碱类成分提出率的比较[J].世界科学技术-中药现代化,2002,4(5):59～77.
[17]郑尚珍,孟军才,等.绿茶中茶多酚的提取工艺研究[M].西北师范大学学报,自然科学版, 1996,32(3):40～45.
[18]李杰红.微波辅助萃取技术提取中药有效成分研究[J].中国医药导报,2006,3(36):155～156.
[19]王晓.酶法提取山楂叶中总黄酮的研究[M].食品工业科技,2003,23(3):37～39.
[20]薛伟明,张效林,亢茂德,等.红花黄色索的酶法提取应用研究[M].化学工程,1999,27(1): 42～46.
[21] Vuataz L, BrandenbergerH, Egli RH. Separation of the tea leaf polyhenols by cellulose column chromatography [J].Chromatographia,1959,2:173～187.
[22]钟世安,周春山,杨娟玉.高效液相色谱分离纯化酯型儿茶素的研究[J].化学世界,2003 (5):237～249.
[23] Silva EM, Pompeu DR, Larondelle Y,et al. Optimisation of the adsorption of polyphenols from Inga edulis leaves on macroporous resins using an experimental design methodology[M]. Separation and Purifation Technology,2007,53(3):274-280.
[24]于曙光.褐藻多酚化合物提取、纯化及生物活性研究[M].青岛:青岛大学化工学院,2003.
[25]龚雨顺,刘仲化,黄建安.茶叶中儿茶素纯化研究进展[J].茶叶通讯,2003(1):15～18.
[26]杜琪珍,李名君,程启坤.高速逆流色谱法分离茶叶中的儿茶素[J].中国茶叶,1996,18(2): 20～21.
[27]林超.两种褐藻中多酚化合物的生理活性研究[D].青岛:青岛大学化工学院,2005.
[28]徐秀丽,范晓,宋福行.中国经济海藻提取物生物活性[J].海洋与湖沼,2004,35(1):55～63.
[29]徐年军,范晓,韩丽君,等.山东沿海海藻抗肿瘤活性的筛选[J].海洋与湖沼,2001,32(4): 408～413.
[30] Harada H.Selective cytotoxicity of marine algae extracts of several human leukemic cell lines [J].Cytotechnology,1997,25(1-3):213～219.
[31] Harada H,Noro T,Kamei Yuto.Selective antitumor activity in vitro from marine algae from Japan coasts[J].Biol Pharm Bull,1997,20(5):541～546.
[32] Yuan YV,Walsh NA.Antioxidant and antiproliferative activities of extracts from a variety of edible seaweeds[J].Food and Chemical Toxicology,2006,44:1144～1150.
[33]牛荣丽,范晓,韩丽君,等.海藻抗A-549和HL-60肿瘤细胞及抗菌活性研究[J].中国海洋药物,2003,22(4):1～4.
[34] Barchi JJ,Moore RE,Patterson GML.Acutiphycin and 20,21-didehydroacutiphycin,new antineoplastic agents from the cyanophyte Oscillatoria acutissima[J].J Am ChemSoc,1984,106:8193～8197.
[35]师然新,徐祖洪.青岛沿海九种海藻的类脂及酚类抗菌活性的研究[J].中国海洋药物,1997,4:16～19.
[36]Hudson JB,Kim JH,Lee MK,et al.Antiviral compounds in extracts of Korean seaweeds: Evidence for multiple activities[J].J Appl Phycol,1999,10: 427～434.
[37]周贞兵.马尾藻多糖与多酚的提取及其活性研究[D].广西:广西大学,2002.
[38] Glombitza KW,Vogels HP.Antibiotics from algae.XXXV.Phlorotannins from Ecklonia maxima[J].Plant Medica,1985,51:308～312.
[39]杨小强.新一代活性饲料——大型海藻饲料[J].饲料研究,2000,1:22～25.
[40]Martinez-Nadal NG,Rodriguez LV,Casillas C.Sarganin and chonalgin,new antiotic substances from marine algae from Puerto Rico[J].Antimicrob Agents Chemother,1964:68～72.
[41]吕志华,于广利,迟连利,等.海黍子多酚对亚油酸甲酯氧化的抑制作用[J].中国海洋药物,2001,6:25～28.
[42] Yan XJ,Li XC,Zhou CX,et al.Prevention of fish oil rancidity by phlorotannins from Sargassum kjllmanianum[J].Appl phycol,1996,8:201～203.
[43]魏玉西,徐祖洪.褐藻中相对高分子质量褐藻多酚的抗氧化活性研究[J].中草药,2003,34(4):317～319.
[44]魏玉西,于曙光.两种褐藻乙醇提取物的抗氧化活性研究[J].海洋科学,2002,26(9):49～51.
[45]范晓,严小军,陈予敏,等.高分子量褐藻多酚抗氧化性质研究[J].水生生物学报,1999,23(5):494～499.
[46] Boettcher AA,Targett NM.Role of polyphenolic molecular size in reduction of asssimilation efficiency in Xiphister mucosus[J].Ecology,1993,4:891～903.
[47] Choi C,Bareiss C,Walenciak O,et al.Impact of polyphenols on growth of the aquatic herbivore Acentria ephemerella[J].J Chem Ecol,2002,28:2245～2256.
[48]曾呈奎,王素娟,刘思俭.海藻栽培学[M].上海:上海科学技术出版社,1985:116.
[49] Chappell J,Hahlbrock.Transcription of plant defense genes in response to UV light or fungal elicitor[J].Nature,1984,311:76～78.
[50] Tokita F,Ishikawa M,Shibuya K,et al.Deodorizing activity of some plant extracts against methyl mercaptan[J].Nippon Nogeikagaku Kaishi,1984,58:585～589.
[51] Kita N,Fujimoto K,Nakajim AI,et al.Screening test for deodorizing substances from marine algae and identification of phlorotannins as the effective ingredients in Eisenia bicyclis[J].J Appl Phycol,1990,2:155～162.
[52] Rajeshkumar NV,Pillai MR,Kuttan R.Induction of apoptosis in mouse and human carcinoma cell lines by Emblica officinalis polyphenols and its effect on chemical carcinogenesis[J].J Exp Clin Cancer Res,2003,22(2):201～212.
[53]裴凌鹏,惠佰隶,金宗濂,等.黄酮类化合物的生理活性及其制备技术研究进展[J].食品科学,2004,25(2):203～207.
[54] Heo SJ, Park EJ,Lee KW,et al.Antioxidant activities of enzymatic extracts from brown seaweeds[J].Bioresour Technol,2005,96(14):1613～1623.
[55]吴勇,陈元仲,许建华,等.姜黄素对人类Burkitt淋巴瘤抗癌作用的研究[J].中华肿瘤杂志,2002,24(4):348～351.
[56] Agarwal C,Singh RP,Agarwal R.Grape seed extract induces apoptotic death of human prostate carcinoma DU145 cells via caspases activation accompanied by dissipation of mitochondrial membrane potential and cytochrome c release[J].Carcinogenesis,2002,23 (11):1869～1876.
[57]陈宏,丁云霞,林旭,等.姜黄素抗大肠癌作用研究[J].肿瘤研究与临床,2006,18(1):4～7.
[58] Feng WH,Wei HL,Liu GT.Effect of PYCNOGENOL on the toxicity of heart,bone marrow and immune organs as induced by antitumor drugs[J].Phytomedicine,2002,9(5):414～418.
[59] Khan SG,Katiyar SK,Agarwal R,et al.Enhacement of antioxidant and phaseⅡenzymes by oral feeding of green tea polyphenols in drinking water to SKH-1 hairless mice :possible role in cancer chemop revention [J].Cancer Res,1992,52(14):4050～4052.
[60] Lin LC,Kuo YC,Chou CJ.Immunomodulatory proanthocyanidins from Ecdysanthera utilis [J].J Nat Prod,2002,65(4):505～508.
[61]曹明富,邵惠琴.茶多酚复合物抑癌及对细胞免疫功能的影响[J].药物生物技术,1999,6 (4):212～217.
[62] Cao Y,Cao R,Brakenhielm E.Antiangiogenic mechanisms of diet-derived polyphenols[J].J Nutr Biochem,2002,13(7):380～390.
[63]Yoo HG,Shin BA,Park JC,et al.Induction of apoptosis by the green tea flavonol(-)-epigallocatechin-3-gallate in human endothelial ECV304 cells[J].Anticancer Res,2002,22(6):3373～3378.
[64] Naasani I,Oh-Hashi F,Oh-Hara T,et al.Blocking telomerase by dietary polyphenols is a major mechanism for limiting the growth of human cancer cells in vitro and in vivo[J].Cancer Res,2003,63(4):824～830.
[65]夏金兰,王春,刘新星.抗菌剂及其抗菌机理[J].中南大学学报(自然科学版),2004,35(1): 31～38.
[66] Ivaneheva S,Manolova N,Serkedjieva J.Polyphenols from Bulgarian dedicinal plants withanti-infections activity[J].Plant polyphenols,1992:621～638.
[67] Schultz J,Hunter M,Appel H.Antimicrobial activity of polyphenols medicines plant herbivore interaction[J].Plant Polyphenols,1992:621～658.
[68]王莹.茶多酚的抗氧化和抑菌活性及其增效剂[J].生物学杂志,2007,24(5):54～56.
[69] Ragan MA,Glombitza KW.Phlorotannins,brown algal polyphenols[J].Progress Phycol, 1986,4:129～241.
[70] Ragan MA,Jensen A.Quantitative studies on brown algal phenols.I.Estimation of absolute polyphenol content of Ascophyllum nodosum (L.) Le Jol.and Fucus vesiculosus (L.)[J].J Exp Mar Biol Ecol,1977,34,245～258.
[71] Swanson AK,Druehl LD.Induction,exudation and the UV protective role of kelp phlorotannins[J].Aquatic Botany,2002,73:241～253.
[72]武文洁,樊文乐,衣守志.海带综合利用研究[J].食品科技,2006(1):49～52.
[73]张铁英.红豆中多酚类物质的提取及其含量测定的研究[J].中国食品添加剂,2004(15):99～100.
[74] Ragan MA,Jensen A.Quantitative studies on brown algal phenols.II.Seasonal variation in polyphenol content of Ascophyllum nodosum (L.) Le Jol.and Fucus vesiculosus (L.)[J].J Exp Mar Biol Ecol,1978,34,245～258.
[75]石碧,狄莹.植物多酚[M].北京:科学出版社,2000.
[76]冯丽,徐桂花,宋曙辉,等.植物多酚及其提取方法的研究进展[J].中国食物与营养,2007(10):39～41.
[77] Jormalainen V,Honkanen T, Vesakoski O,et al.Polar extracts of the brown alga Fucus vesiculosus (L.) reduce assimilation efficiency but do not deter the herbivorous isopod Idotea baltica (Pallas)[J].Exp Mar Biol Ecol,2005,317:143～157.
[78] Vaher M,Koel M.Separation of polyphenolic compounds extracted from plant matrices using capillary electrophoresis[J].J Chromatogr A,2003,990:225～230.
[79] Pekic B,Kovac V,Alonso E.Study of the extraction of proanthocyanidins from grape Seeds[J].Food Chem,1998,61(12):201～206.
[80]李华.超声波法从葡萄籽中提取多酚的研究[J].酿酒科技,2005,(5):89～91.
[81] Mossman T.Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assay[J].Journal of Immunological Methods,1983,65:55～63.
[82] Athukorala Y, Kim KN, Jeon YJ.Antiproliferative and antioxidant properties of an enzymatic hydrolysate from brown alga,Ecklonia cava[J].Food and Chemical Toxicology,2006,44: 1065～1074.
[83]莫晓媚,李静,李延团,耿美玉.一种钒配合物LMC抑制DNA拓扑异构酶Ⅰ的抗肿瘤作用[J].现代生物医学进展,2006,4(6):13～15.
[84] Haugen A.Molecular biology in diagnosis of lung cancer[J].Tidsskr Nor Laegeforen, 2005,125(23):3283～3285.
[85] Granville CA,Dennis PA.An overview of lung caneer genomics and proteomics[J].Am J Respir Cell Mol Biol,2005,32(3):169～176.
[86] Harris N,Jaffe ES,Diebold T,et al.World Health 0rganization classification of neoplastic diseases of the hematopoietic and lymphoid tissues: report of the clinical advisory committee meeting-Airlie house,Virgina,November 1997.J Clin Oncol,1999,17:3835～3849.
[87] Parkin DM,Bray F,Ferlay J,et al.Global caneer statistics,2002[J].CA Cancer J Clin,2005,55:74～108.
[88] Chen CJ,Yang Hl,Su J, et al.Risk of hepatocellular carcinoma across a biological gradient of serum hepatitis B virus DNA level[J].JAMA.2006,295:65～73.
[89] Boyle P,Veeehia C,Walker A.FIGO annual report on the results of treatment in gynecological cancer[J].Journal of epidemiology and biostatistics ,2001,12:7～43.
[90]林超,于曙光,郭道森,等.鼠尾藻中褐藻多酚化合物的抑菌活性研究[J].海洋科学,2006,30(3):94～97.
[91]裘爱泳,刘军海,张海晖.植物多酚提取和应用[J].粮食与油脂,2003,(6):10～11.
[92] Caccamese S,ToscanaR M,Furnari G,et al. Antimicrobial activities of red and brown algae from Southern Italy Coast[J].Bot Mar,1985, 28:505～507.
[93] Pesando D,Caram B.Screening of marine algae from the French Mediterranean Coast for antibacterial and antifungal activity[J].Bot Mar,1984,27:381～386.
[94] Reichelt JL,Borowizka MA.Antimicrobial activity from marine algae: Results of a large scale screening programme[J].Hydrobiologia,1984,22:158～167.
[95] Rao PPS,Rao PS,Karmarkar SM.Antibacterial substances from brow algae[J].Bot Mar, 1986,29:503～507.
[96]Hodgsobn LM.Antimicrobial and antineoplastic activity in some south Florida seameeds[J]. Bot Mar,1984,27:387～390.
[97]蔡杉杉.中药抗深部真菌实验研究进展[J].中医函授通讯,2000,19(4):11～14.
[98]王宏.苹果渣中多酚物质的提取分离及其抗氧化活性研究[D].陕西:陕西师范大学食品科学,2006.
[99] Paola P,Rita M,Sonia T,et al.Solubilization and stabilization ofβ-carotene in niosomes:delivery to cultured cells[J].Chemistry and Physics of Lipids,2006,139(1):32～42.
[100]宁永成.有机化合物结构鉴定与有机波谱学[M].北京:科学出版社,2000:364～384.
[101] Swanson AK,Druehl LD.Induction,exudation and the UV protective role of kelpphlorotannins[J].Aquatic Botany,2002,73:241～253.
[102] Pavia H,Cervin G, Lindgren A, Aberg P. The effect of UV-B radiation and simulated herbivory on the production of phlorotannins in the brown seaweed Ascophyllum nodosum[J]. Marine Ecology Progress Series,1997,157:139～146.
[103]李述文,范如霖.实用有机化学手册[M].上海:上海科学技术出版社,1981.
[104]贾之慎.比色法测定Fenton反应产生的经自由基[J].生物化学与生物物理进展,1996,23(2):184～186.
[105]王春波,贺孟泉,秦守哲.海洋肽的体外抗氧化作用[J],中国海洋药物,1998,3(67):15～17.
[106]郭丽萍.香蕉皮中多酚物质抗氧化性能的研究[D].广西:广西大学,2006.
[107]谢样茂,丁小雯,陈俊琴.金樱子提取液对NO2-清除作用的体外实验研究[J].食品科学, 2001,2(1):30～33.
[108]褚衍信,冯华进,孙燕.亚硝酸根清除剂体外研究的新方法及其应用[J].癌症,1994,5:470～471.
[109]郝会芳.枣果中多酚类物质的提取工艺及体外抗氧化作用研究[D].河北:河北农业大学, 2005.
[110] Cui CB,Yan SY,Cai B,et al.Carbazole alkaloids as new cell cycle inhibitors and apoptosis inducers from Clausena DunnianaLevl[J].J Asian Nat Prod Res,2002,4(4): 233～241.
[111] Cui CB,Zhao QC,Cai B,et al.Two new and four known polyphenolics obtained as new cell-cycle inhibitors fromRubus aleaefolius Poir [J].J Asian Nat Prod Res,2002,4(4):243～252.
[112]郑铁生.菌霉克星防霉剂抗菌效力和机理的研究[J].镇江医学院学报,1999,9(2):199～200.
[113]杨贤强,詹皓,沈生荣,等.茶多酚对脑损伤的保护作用[J].茶叶科学,1997,(17):124～130.
[2] Handley JT,Blackman AJ.Secondary metabolites from the marine alga Caulerpa brownii (Chlorophyta)[J].Australian Journal of Chemistry,2005,58(1):39～46.
[3] Erickson AA,Paul VJ,Van Alstyne KL,Kwiatkowski LM.Palatability of macroalgae that use different types of chemical defenses[J].Journal of Chemical Ecology,2006,31(9):1883～1895.
[4]纪明侯.海藻化学[M].北京:科学出版社,1997:628.
[5]徐秀丽,范晓,宋福行,等.褐藻小粘膜藻中的溴酚化合物[J].中国中药杂志,2005,30(1):39～41.
[6]严小军.中国常见褐藻的多酚含量测定[J].海洋科学集刊,1996,37:61～64.
[7] Kanget.Study on the extraction condition and quantity of polyphenols in a dry brown algae Ecklonia cava[J].Bull Mar Res,1990,14:25～38.
[8]Barwell CJ,Blunden G,Manandhar PD.Isolation and characterization of brown algal polyphenols as inhibitors ofα-amylase,lipase,and trypsin [J].J Appl Phycol,1989,1:319-323.
[9] Keusgen M,Glombitza KW.Phlorethols,fuhalols,and their derivatives from the brown alga Sargassum spinulligerum[J].Phytochemistry,1995,38(4):975～985.
[10]Glombitza KW,Keusgen M,Hauperich S.Fucophlorethols from the algae Sargassum spinuligerum and Cystophora torulosa[J].Phytochemistry,1997,46(8):1417～1422.
[11] Sailler B,Glombitza KW.Phlorethols and fucophlorethols from the brown alga Cystophora retroflexa[J].Phytochemistry,1999,50:869～881.
[12]Glombitza KW,Schmidt A.Trihydroxyphlorethols from the brown alga Carpophyllum angustifolium[J].Phytochemistry,1999,51:1095～1100.
[13] Smller B,Glombitza KW.Halogenated phlorethols and fucophlorethols from the brown alga Cystophora retroflexa[J]. Nat Toxins,1999,7(2):57～62.
[14] Glombitza KW,Schmidt A.Nonhalogenated and halogenated phlorotannins from the brown alga Carpophyllum angustifolium[J].J Nat Prod,1999,62(9):1238～1240.
[15]陈予敏,严小军,范晓.褐藻多酚的结构单元及生成机理[J].海洋与湖沼,1997,28(3):225～231.
[16]郭孝武.超声提取与常规提取对部分中药碱类成分提出率的比较[J].世界科学技术-中药现代化,2002,4(5):59～77.
[17]郑尚珍,孟军才,等.绿茶中茶多酚的提取工艺研究[M].西北师范大学学报,自然科学版, 1996,32(3):40～45.
[18]李杰红.微波辅助萃取技术提取中药有效成分研究[J].中国医药导报,2006,3(36):155～156.
[19]王晓.酶法提取山楂叶中总黄酮的研究[M].食品工业科技,2003,23(3):37～39.
[20]薛伟明,张效林,亢茂德,等.红花黄色索的酶法提取应用研究[M].化学工程,1999,27(1): 42～46.
[21] Vuataz L, BrandenbergerH, Egli RH. Separation of the tea leaf polyhenols by cellulose column chromatography [J].Chromatographia,1959,2:173～187.
[22]钟世安,周春山,杨娟玉.高效液相色谱分离纯化酯型儿茶素的研究[J].化学世界,2003 (5):237～249.
[23] Silva EM, Pompeu DR, Larondelle Y,et al. Optimisation of the adsorption of polyphenols from Inga edulis leaves on macroporous resins using an experimental design methodology[M]. Separation and Purifation Technology,2007,53(3):274-280.
[24]于曙光.褐藻多酚化合物提取、纯化及生物活性研究[M].青岛:青岛大学化工学院,2003.
[25]龚雨顺,刘仲化,黄建安.茶叶中儿茶素纯化研究进展[J].茶叶通讯,2003(1):15～18.
[26]杜琪珍,李名君,程启坤.高速逆流色谱法分离茶叶中的儿茶素[J].中国茶叶,1996,18(2): 20～21.
[27]林超.两种褐藻中多酚化合物的生理活性研究[D].青岛:青岛大学化工学院,2005.
[28]徐秀丽,范晓,宋福行.中国经济海藻提取物生物活性[J].海洋与湖沼,2004,35(1):55～63.
[29]徐年军,范晓,韩丽君,等.山东沿海海藻抗肿瘤活性的筛选[J].海洋与湖沼,2001,32(4): 408～413.
[30] Harada H.Selective cytotoxicity of marine algae extracts of several human leukemic cell lines [J].Cytotechnology,1997,25(1-3):213～219.
[31] Harada H,Noro T,Kamei Yuto.Selective antitumor activity in vitro from marine algae from Japan coasts[J].Biol Pharm Bull,1997,20(5):541～546.
[32] Yuan YV,Walsh NA.Antioxidant and antiproliferative activities of extracts from a variety of edible seaweeds[J].Food and Chemical Toxicology,2006,44:1144～1150.
[33]牛荣丽,范晓,韩丽君,等.海藻抗A-549和HL-60肿瘤细胞及抗菌活性研究[J].中国海洋药物,2003,22(4):1～4.
[34] Barchi JJ,Moore RE,Patterson GML.Acutiphycin and 20,21-didehydroacutiphycin,new antineoplastic agents from the cyanophyte Oscillatoria acutissima[J].J Am ChemSoc,1984,106:8193～8197.
[35]师然新,徐祖洪.青岛沿海九种海藻的类脂及酚类抗菌活性的研究[J].中国海洋药物,1997,4:16～19.
[36]Hudson JB,Kim JH,Lee MK,et al.Antiviral compounds in extracts of Korean seaweeds: Evidence for multiple activities[J].J Appl Phycol,1999,10: 427～434.
[37]周贞兵.马尾藻多糖与多酚的提取及其活性研究[D].广西:广西大学,2002.
[38] Glombitza KW,Vogels HP.Antibiotics from algae.XXXV.Phlorotannins from Ecklonia maxima[J].Plant Medica,1985,51:308～312.
[39]杨小强.新一代活性饲料——大型海藻饲料[J].饲料研究,2000,1:22～25.
[40]Martinez-Nadal NG,Rodriguez LV,Casillas C.Sarganin and chonalgin,new antiotic substances from marine algae from Puerto Rico[J].Antimicrob Agents Chemother,1964:68～72.
[41]吕志华,于广利,迟连利,等.海黍子多酚对亚油酸甲酯氧化的抑制作用[J].中国海洋药物,2001,6:25～28.
[42] Yan XJ,Li XC,Zhou CX,et al.Prevention of fish oil rancidity by phlorotannins from Sargassum kjllmanianum[J].Appl phycol,1996,8:201～203.
[43]魏玉西,徐祖洪.褐藻中相对高分子质量褐藻多酚的抗氧化活性研究[J].中草药,2003,34(4):317～319.
[44]魏玉西,于曙光.两种褐藻乙醇提取物的抗氧化活性研究[J].海洋科学,2002,26(9):49～51.
[45]范晓,严小军,陈予敏,等.高分子量褐藻多酚抗氧化性质研究[J].水生生物学报,1999,23(5):494～499.
[46] Boettcher AA,Targett NM.Role of polyphenolic molecular size in reduction of asssimilation efficiency in Xiphister mucosus[J].Ecology,1993,4:891～903.
[47] Choi C,Bareiss C,Walenciak O,et al.Impact of polyphenols on growth of the aquatic herbivore Acentria ephemerella[J].J Chem Ecol,2002,28:2245～2256.
[48]曾呈奎,王素娟,刘思俭.海藻栽培学[M].上海:上海科学技术出版社,1985:116.
[49] Chappell J,Hahlbrock.Transcription of plant defense genes in response to UV light or fungal elicitor[J].Nature,1984,311:76～78.
[50] Tokita F,Ishikawa M,Shibuya K,et al.Deodorizing activity of some plant extracts against methyl mercaptan[J].Nippon Nogeikagaku Kaishi,1984,58:585～589.
[51] Kita N,Fujimoto K,Nakajim AI,et al.Screening test for deodorizing substances from marine algae and identification of phlorotannins as the effective ingredients in Eisenia bicyclis[J].J Appl Phycol,1990,2:155～162.
[52] Rajeshkumar NV,Pillai MR,Kuttan R.Induction of apoptosis in mouse and human carcinoma cell lines by Emblica officinalis polyphenols and its effect on chemical carcinogenesis[J].J Exp Clin Cancer Res,2003,22(2):201～212.
[53]裴凌鹏,惠佰隶,金宗濂,等.黄酮类化合物的生理活性及其制备技术研究进展[J].食品科学,2004,25(2):203～207.
[54] Heo SJ, Park EJ,Lee KW,et al.Antioxidant activities of enzymatic extracts from brown seaweeds[J].Bioresour Technol,2005,96(14):1613～1623.
[55]吴勇,陈元仲,许建华,等.姜黄素对人类Burkitt淋巴瘤抗癌作用的研究[J].中华肿瘤杂志,2002,24(4):348～351.
[56] Agarwal C,Singh RP,Agarwal R.Grape seed extract induces apoptotic death of human prostate carcinoma DU145 cells via caspases activation accompanied by dissipation of mitochondrial membrane potential and cytochrome c release[J].Carcinogenesis,2002,23 (11):1869～1876.
[57]陈宏,丁云霞,林旭,等.姜黄素抗大肠癌作用研究[J].肿瘤研究与临床,2006,18(1):4～7.
[58] Feng WH,Wei HL,Liu GT.Effect of PYCNOGENOL on the toxicity of heart,bone marrow and immune organs as induced by antitumor drugs[J].Phytomedicine,2002,9(5):414～418.
[59] Khan SG,Katiyar SK,Agarwal R,et al.Enhacement of antioxidant and phaseⅡenzymes by oral feeding of green tea polyphenols in drinking water to SKH-1 hairless mice :possible role in cancer chemop revention [J].Cancer Res,1992,52(14):4050～4052.
[60] Lin LC,Kuo YC,Chou CJ.Immunomodulatory proanthocyanidins from Ecdysanthera utilis [J].J Nat Prod,2002,65(4):505～508.
[61]曹明富,邵惠琴.茶多酚复合物抑癌及对细胞免疫功能的影响[J].药物生物技术,1999,6 (4):212～217.
[62] Cao Y,Cao R,Brakenhielm E.Antiangiogenic mechanisms of diet-derived polyphenols[J].J Nutr Biochem,2002,13(7):380～390.
[63]Yoo HG,Shin BA,Park JC,et al.Induction of apoptosis by the green tea flavonol(-)-epigallocatechin-3-gallate in human endothelial ECV304 cells[J].Anticancer Res,2002,22(6):3373～3378.
[64] Naasani I,Oh-Hashi F,Oh-Hara T,et al.Blocking telomerase by dietary polyphenols is a major mechanism for limiting the growth of human cancer cells in vitro and in vivo[J].Cancer Res,2003,63(4):824～830.
[65]夏金兰,王春,刘新星.抗菌剂及其抗菌机理[J].中南大学学报(自然科学版),2004,35(1): 31～38.
[66] Ivaneheva S,Manolova N,Serkedjieva J.Polyphenols from Bulgarian dedicinal plants withanti-infections activity[J].Plant polyphenols,1992:621～638.
[67] Schultz J,Hunter M,Appel H.Antimicrobial activity of polyphenols medicines plant herbivore interaction[J].Plant Polyphenols,1992:621～658.
[68]王莹.茶多酚的抗氧化和抑菌活性及其增效剂[J].生物学杂志,2007,24(5):54～56.
[69] Ragan MA,Glombitza KW.Phlorotannins,brown algal polyphenols[J].Progress Phycol, 1986,4:129～241.
[70] Ragan MA,Jensen A.Quantitative studies on brown algal phenols.I.Estimation of absolute polyphenol content of Ascophyllum nodosum (L.) Le Jol.and Fucus vesiculosus (L.)[J].J Exp Mar Biol Ecol,1977,34,245～258.
[71] Swanson AK,Druehl LD.Induction,exudation and the UV protective role of kelp phlorotannins[J].Aquatic Botany,2002,73:241～253.
[72]武文洁,樊文乐,衣守志.海带综合利用研究[J].食品科技,2006(1):49～52.
[73]张铁英.红豆中多酚类物质的提取及其含量测定的研究[J].中国食品添加剂,2004(15):99～100.
[74] Ragan MA,Jensen A.Quantitative studies on brown algal phenols.II.Seasonal variation in polyphenol content of Ascophyllum nodosum (L.) Le Jol.and Fucus vesiculosus (L.)[J].J Exp Mar Biol Ecol,1978,34,245～258.
[75]石碧,狄莹.植物多酚[M].北京:科学出版社,2000.
[76]冯丽,徐桂花,宋曙辉,等.植物多酚及其提取方法的研究进展[J].中国食物与营养,2007(10):39～41.
[77] Jormalainen V,Honkanen T, Vesakoski O,et al.Polar extracts of the brown alga Fucus vesiculosus (L.) reduce assimilation efficiency but do not deter the herbivorous isopod Idotea baltica (Pallas)[J].Exp Mar Biol Ecol,2005,317:143～157.
[78] Vaher M,Koel M.Separation of polyphenolic compounds extracted from plant matrices using capillary electrophoresis[J].J Chromatogr A,2003,990:225～230.
[79] Pekic B,Kovac V,Alonso E.Study of the extraction of proanthocyanidins from grape Seeds[J].Food Chem,1998,61(12):201～206.
[80]李华.超声波法从葡萄籽中提取多酚的研究[J].酿酒科技,2005,(5):89～91.
[81] Mossman T.Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assay[J].Journal of Immunological Methods,1983,65:55～63.
[82] Athukorala Y, Kim KN, Jeon YJ.Antiproliferative and antioxidant properties of an enzymatic hydrolysate from brown alga,Ecklonia cava[J].Food and Chemical Toxicology,2006,44: 1065～1074.
[83]莫晓媚,李静,李延团,耿美玉.一种钒配合物LMC抑制DNA拓扑异构酶Ⅰ的抗肿瘤作用[J].现代生物医学进展,2006,4(6):13～15.
[84] Haugen A.Molecular biology in diagnosis of lung cancer[J].Tidsskr Nor Laegeforen, 2005,125(23):3283～3285.
[85] Granville CA,Dennis PA.An overview of lung caneer genomics and proteomics[J].Am J Respir Cell Mol Biol,2005,32(3):169～176.
[86] Harris N,Jaffe ES,Diebold T,et al.World Health 0rganization classification of neoplastic diseases of the hematopoietic and lymphoid tissues: report of the clinical advisory committee meeting-Airlie house,Virgina,November 1997.J Clin Oncol,1999,17:3835～3849.
[87] Parkin DM,Bray F,Ferlay J,et al.Global caneer statistics,2002[J].CA Cancer J Clin,2005,55:74～108.
[88] Chen CJ,Yang Hl,Su J, et al.Risk of hepatocellular carcinoma across a biological gradient of serum hepatitis B virus DNA level[J].JAMA.2006,295:65～73.
[89] Boyle P,Veeehia C,Walker A.FIGO annual report on the results of treatment in gynecological cancer[J].Journal of epidemiology and biostatistics ,2001,12:7～43.
[90]林超,于曙光,郭道森,等.鼠尾藻中褐藻多酚化合物的抑菌活性研究[J].海洋科学,2006,30(3):94～97.
[91]裘爱泳,刘军海,张海晖.植物多酚提取和应用[J].粮食与油脂,2003,(6):10～11.
[92] Caccamese S,ToscanaR M,Furnari G,et al. Antimicrobial activities of red and brown algae from Southern Italy Coast[J].Bot Mar,1985, 28:505～507.
[93] Pesando D,Caram B.Screening of marine algae from the French Mediterranean Coast for antibacterial and antifungal activity[J].Bot Mar,1984,27:381～386.
[94] Reichelt JL,Borowizka MA.Antimicrobial activity from marine algae: Results of a large scale screening programme[J].Hydrobiologia,1984,22:158～167.
[95] Rao PPS,Rao PS,Karmarkar SM.Antibacterial substances from brow algae[J].Bot Mar, 1986,29:503～507.
[96]Hodgsobn LM.Antimicrobial and antineoplastic activity in some south Florida seameeds[J]. Bot Mar,1984,27:387～390.
[97]蔡杉杉.中药抗深部真菌实验研究进展[J].中医函授通讯,2000,19(4):11～14.
[98]王宏.苹果渣中多酚物质的提取分离及其抗氧化活性研究[D].陕西:陕西师范大学食品科学,2006.
[99] Paola P,Rita M,Sonia T,et al.Solubilization and stabilization ofβ-carotene in niosomes:delivery to cultured cells[J].Chemistry and Physics of Lipids,2006,139(1):32～42.
[100]宁永成.有机化合物结构鉴定与有机波谱学[M].北京:科学出版社,2000:364～384.
[101] Swanson AK,Druehl LD.Induction,exudation and the UV protective role of kelpphlorotannins[J].Aquatic Botany,2002,73:241～253.
[102] Pavia H,Cervin G, Lindgren A, Aberg P. The effect of UV-B radiation and simulated herbivory on the production of phlorotannins in the brown seaweed Ascophyllum nodosum[J]. Marine Ecology Progress Series,1997,157:139～146.
[103]李述文,范如霖.实用有机化学手册[M].上海:上海科学技术出版社,1981.
[104]贾之慎.比色法测定Fenton反应产生的经自由基[J].生物化学与生物物理进展,1996,23(2):184～186.
[105]王春波,贺孟泉,秦守哲.海洋肽的体外抗氧化作用[J],中国海洋药物,1998,3(67):15～17.
[106]郭丽萍.香蕉皮中多酚物质抗氧化性能的研究[D].广西:广西大学,2006.
[107]谢样茂,丁小雯,陈俊琴.金樱子提取液对NO2-清除作用的体外实验研究[J].食品科学, 2001,2(1):30～33.
[108]褚衍信,冯华进,孙燕.亚硝酸根清除剂体外研究的新方法及其应用[J].癌症,1994,5:470～471.
[109]郝会芳.枣果中多酚类物质的提取工艺及体外抗氧化作用研究[D].河北:河北农业大学, 2005.
[110] Cui CB,Yan SY,Cai B,et al.Carbazole alkaloids as new cell cycle inhibitors and apoptosis inducers from Clausena DunnianaLevl[J].J Asian Nat Prod Res,2002,4(4): 233～241.
[111] Cui CB,Zhao QC,Cai B,et al.Two new and four known polyphenolics obtained as new cell-cycle inhibitors fromRubus aleaefolius Poir [J].J Asian Nat Prod Res,2002,4(4):243～252.
[112]郑铁生.菌霉克星防霉剂抗菌效力和机理的研究[J].镇江医学院学报,1999,9(2):199～200.
[113]杨贤强,詹皓,沈生荣,等.茶多酚对脑损伤的保护作用[J].茶叶科学,1997,(17):124～130.