芥子碱对小鼠肝癌H22的抑制作用及其机制初探
|
摘要
原发性肝癌是恶性肿瘤之首,调查显示我国每年死于肝癌的人数超过11万,占世界肝癌死亡人数的45%。在临床抗肿瘤药物研究中,源于植物的生物碱一直都是抗肿瘤药物的重要来源。近年来,生物碱类抗肿瘤作用及其抗肿瘤机制的研究已成为攻克肿瘤这一世纪难题的重要领域之一。但目前临床上广泛运用的生物碱类药物均存在毒副作用强、耐药性高、选择性差的缺点,能否找到一种低毒、高效、选择性好的抗肿瘤药物已成为国内外学者研究的热点。
本研究通过分离、纯化白芥子的主要药效成分——芥子碱,研究其对小鼠肝癌H22的抑制作用及其机制,为芥子碱的开发应用提供参考。
采用溶剂萃取法分离制备芥子碱,以乙醇与水多次交互重结晶,得到浅黄色针状结晶芥子碱,得率为0.382%,并采用高效液相色谱法和分光光度法测定其纯度达95%以上。
通过MTT法和细胞排染法检测芥子碱对小鼠肝癌H22细胞体外增殖的影响,并通过DNA Ladder试验检测是否发生凋亡。结果显示：芥子碱对小鼠肝癌H22细胞增殖有显著的体外抑制作用,且给药组出现不同程度的凋亡特征现象。
昆明小鼠接种H22细胞,分别建立腹水型肝癌H22模型和小鼠肝癌H22实体瘤模型,观察芥子碱对小鼠肝癌H22的体内抑制作用,HE染色法观察细胞形态。通过检测胸腺指数、脾脏指数探究芥子碱对小鼠免疫器官的影响,并采用丫啶橙染色观察细胞凋亡形态,琼脂糖凝胶电泳及免疫组织化学法研究芥子碱的诱导细胞凋亡机制。实验结果显示：芥子碱对小鼠肝癌H22实体瘤有显著的抑制作用,存在剂量依赖效应,且未出现明显毒副作用,而对腹水型H22小鼠的生命延长无显著改善。凋亡机制探究发现芥子碱可以促进促凋亡因子Bax表达、抑制抗凋亡因子Bcl-2的表达。
Primary liver cancer(PLC) is one of the most common malignant tumor, which threats seriously to human health and life. Statistics showed that PLC kills an estimated 110 000 people, or forty-five percent of total global deaths each year. Alkaloid extracted from plants is always a very important resources to anti-tumor drugs. However, the drugs that were used extensively in clinic have some defects, such as high toxicity, poor specificity. The development of low toxicity, high efficiency, and good specificity of anti-tumor drugs has become a hot issue in all over the world.
In this paper, sinapine was separated from seeds of Brassica alba. The in vitro and in vivo inhibitory effect of sinapine on H22 was studied. And the mechanism of anti-tumor effect of sinapine on H22 hepatoma-bearing mice was explored. These may be the good basis for further study on sinapine and looking for safe and effective anti-tumor drugs.
Seeds of Brassica alba were smashed, defatted by ether, extracted by alcohol, and crystallized by potassium thiocyanate, then ethanol and water were used to recrystallize on many occasions. The yield of sinapine was 0.382%. The determination was performed by reversed-phase high performance liquid chromatography (RP-HPLC) and ultraviolet spectrophotometry (UV Spectrophotometry).
MTT assay and Trypan blue staining was used to detect the inhibitionrate of H22 hepatocarcinoma cells by sinapine in vitro. And the DNA changes of H22 hepatocarcinoma cells (actived by sinapine) were observed by DNA Ladder. The results showed that sinapine could obviously inhibited the proliferation of H22 hepatocarcinoma cells, and the phenomenon of apoptosis of H22 hepatocarcinoma cells (actived by sinapine) was observed.
Subcutaneous injection of H22 hepatocarcinoma cells to mice, creating the mice models of H22. The anti-tumor effect of sinapine was evaluated by observation of rate of tumor inhibition and immunity index. The pathological changes in tumor tissue was observed by HE staining. AO staining, Agarose gel electrophoresis analysis and Immunohisto-chemical assay were used to explore the possible mechanism of induction of apoptosis.
The results suggested that the growth of H22 solid tumor in vivo were significantly inhibited by sinapine without toxicity, and the concentration increases with the dose effect relationship. But the effect of sinapine on survivor life of mice bearing H22 ascites tumor is not significant. The immunohisto-chemical results showed that sinapine can enhance the expression of Bax protein and reduced Bcl-2 protein expression.
本研究通过分离、纯化白芥子的主要药效成分——芥子碱,研究其对小鼠肝癌H22的抑制作用及其机制,为芥子碱的开发应用提供参考。
采用溶剂萃取法分离制备芥子碱,以乙醇与水多次交互重结晶,得到浅黄色针状结晶芥子碱,得率为0.382%,并采用高效液相色谱法和分光光度法测定其纯度达95%以上。
通过MTT法和细胞排染法检测芥子碱对小鼠肝癌H22细胞体外增殖的影响,并通过DNA Ladder试验检测是否发生凋亡。结果显示：芥子碱对小鼠肝癌H22细胞增殖有显著的体外抑制作用,且给药组出现不同程度的凋亡特征现象。
昆明小鼠接种H22细胞,分别建立腹水型肝癌H22模型和小鼠肝癌H22实体瘤模型,观察芥子碱对小鼠肝癌H22的体内抑制作用,HE染色法观察细胞形态。通过检测胸腺指数、脾脏指数探究芥子碱对小鼠免疫器官的影响,并采用丫啶橙染色观察细胞凋亡形态,琼脂糖凝胶电泳及免疫组织化学法研究芥子碱的诱导细胞凋亡机制。实验结果显示：芥子碱对小鼠肝癌H22实体瘤有显著的抑制作用,存在剂量依赖效应,且未出现明显毒副作用,而对腹水型H22小鼠的生命延长无显著改善。凋亡机制探究发现芥子碱可以促进促凋亡因子Bax表达、抑制抗凋亡因子Bcl-2的表达。
Primary liver cancer(PLC) is one of the most common malignant tumor, which threats seriously to human health and life. Statistics showed that PLC kills an estimated 110 000 people, or forty-five percent of total global deaths each year. Alkaloid extracted from plants is always a very important resources to anti-tumor drugs. However, the drugs that were used extensively in clinic have some defects, such as high toxicity, poor specificity. The development of low toxicity, high efficiency, and good specificity of anti-tumor drugs has become a hot issue in all over the world.
In this paper, sinapine was separated from seeds of Brassica alba. The in vitro and in vivo inhibitory effect of sinapine on H22 was studied. And the mechanism of anti-tumor effect of sinapine on H22 hepatoma-bearing mice was explored. These may be the good basis for further study on sinapine and looking for safe and effective anti-tumor drugs.
Seeds of Brassica alba were smashed, defatted by ether, extracted by alcohol, and crystallized by potassium thiocyanate, then ethanol and water were used to recrystallize on many occasions. The yield of sinapine was 0.382%. The determination was performed by reversed-phase high performance liquid chromatography (RP-HPLC) and ultraviolet spectrophotometry (UV Spectrophotometry).
MTT assay and Trypan blue staining was used to detect the inhibitionrate of H22 hepatocarcinoma cells by sinapine in vitro. And the DNA changes of H22 hepatocarcinoma cells (actived by sinapine) were observed by DNA Ladder. The results showed that sinapine could obviously inhibited the proliferation of H22 hepatocarcinoma cells, and the phenomenon of apoptosis of H22 hepatocarcinoma cells (actived by sinapine) was observed.
Subcutaneous injection of H22 hepatocarcinoma cells to mice, creating the mice models of H22. The anti-tumor effect of sinapine was evaluated by observation of rate of tumor inhibition and immunity index. The pathological changes in tumor tissue was observed by HE staining. AO staining, Agarose gel electrophoresis analysis and Immunohisto-chemical assay were used to explore the possible mechanism of induction of apoptosis.
The results suggested that the growth of H22 solid tumor in vivo were significantly inhibited by sinapine without toxicity, and the concentration increases with the dose effect relationship. But the effect of sinapine on survivor life of mice bearing H22 ascites tumor is not significant. The immunohisto-chemical results showed that sinapine can enhance the expression of Bax protein and reduced Bcl-2 protein expression.
引文
[1]Pelletier, S. Alkaloids:Chenmical and Biological Perspectives[M]. New York:Jhon Wiley & Sons, 1983:26
[2]Wani M, Taylor H, Wall M, et al. Plant antitumor agents. VI. The isolation and structure of taxol, a novel antileukemic and antitumor agent from Taxus brevifolia[J]. Am Chem Soc,1971,93(9): 2325-2327
[3]张骁,束梅英,张韬.紫杉醇的研究进展[J].中国医药技术与市场,2006,6(4)：40-46
[4]Stone R. Surprise! A fungus factory for taxol[J]? Science,1993,260(5150):154-155
[5]Markman M, Hall J, Spitz DW, et al. Phase II trial of weekly single-agent paclitaxel in platinum/paclitaxel-refractory ovarian cancer[J]. Clin Oncol,2002,20(9):2365-2369
[6]Alberto M, Frank F, Gabriel H, et al. Docetaxel for treatment of solid tumours:a systenmatic review of clinical data. Lancet Oncol,2005,6:229-239
[7]AL-Sarrf M. Treatment of locally advanced head and neck cancer[J]. Cancer Contr,2002,9(5): 387-399
[8]Schiff P, Fant J, Horwitz S. Promation of microtubule assembly in vitro by taxol. Nature 1979, 277(5698):665-667
[9]Scatena C, Stewart Z, Mays D. Mitotic phosphorylation of Bcl-2 during normal cell cycle progression and Paclitaxel-induced growth arest[J]. Biol Chem.1998,273(46):30777-30784
[10]Haldar S, Basu A, Croce C. Serine-70 is one of the critical sites for drug-induced Bcl-2 phosphorylation in cancer cells. Cancer Res.1998,58(8):1609-1615
[11]Panvichian R, Orth K, Day ML, et al. Paclitaxel-associated multiminricmcleation is permitted by the inhibition og caspas sctivation:a potential early step in druy resistance[J]. Cancer Res,1998, 58(20):4667
[12]孟刚,李庆林,陈志武,等.紫杉醇诱导MCF27凋亡与bcl-2及bax的关系[J].中国药理学通报,2001,17(3)：282-285
[13]Yoshino T, Shiina H, Urakami S, et al. Bcl-2 expression as a predictive marker of hormone-refractory prostate cancer treated with taxane-based chemotherapy. Clin Cancer Res, 2006,238(1):90-103
[14]Blagosklonny M, Schulte T, Nguyen P, et al. Taxol-induced apoptosis and phosphorylation of Bcl-2 protein involves c-Raf-1 and represents a novel c-Raf-1 signal transduction pathway[J]. Cancer Res,1996,56(8):1851-1854
[15]Uffelie OF. Alkaloids with tumor inhibiting action from Catharanthus roseus[J]. Pharm Weekbl, 1969,104(16):321-330
[16]Zhou X, Ranmani R. Preclinical pharmacology of vinca alkaloids[J]. Drugs,1992,44(4):1-16
[17]丁亚芳,包永明,安利佳.长春碱类抗肿瘤药物的研究进展[J].中国医药工业杂志,2005,36(7)：424-427
[18]Ngan VK, Bellman K, Panda D, et al. Novel actions of the antitumor drugs vinflunine and vinorelbine on microtubules[J], Cancer Res,2000,60:5045-5051
[19]Lobert S, Vulevic B, Correia J. Interaction of vinca alkaloids with tubulin:a comparison of vinblastine, vincristine, and vinorelbine[J]. Biochem,1996,35(21):6806-6814
[20]马彦.长春碱类抗肿瘤药物的研究进展.中山大学研究生学刊,2004,25(3)：30-39
[21]Pourroy B, Carre M, Honore S, et al. Low concentration of vinflunine induce apoptosis in human SK-N-SH neuroblastoma cells through a postmitotic G1 arrest and a mitochondrial pathway[J]. Mol Pharmacol,2004,66(3):580-591
[22]Azra R, Elham C, Zahra H. The effect of vinca alkaloid anticancer drug, vinorelbine, on chromatin and histone proteins in solution[J]. Eur. J. Pharmac,2009,613(1-3):34-38
[23]Yuan J, Zhang Z. Advances research and Bio-activity of matrine[J]. Pesticides,2003,42(7):1-4
[24]Wang T, Li s, Jian J, et al. Advances in the studies of anti-tumour activity of alkaloids in Radix Sophorae Flacescentis (Kushen)[J]. Chin J Exp Trad Med Form,2004,10(4):52-55
[25]Jin Y, Wu X, Lou J. Effects of matrine on cell proliferation, cell cycle and apoptosis of human hepatocarcinoma cells[J]. Chin. J. Clin Rehab.2006,10(3):107-109
[26]张鸣杰,黄健.苦参碱类抗肿瘤作用机制研究的新进展[J].中国中药杂志,2004,29(2)：115-118
[27]Dai Z, Gao J, Ji Z, et al. Matrine induces apoptosis in gastric carcinoma cells via alteration of Fas/FasL and activation of caspase-3[J]. J Ethnophrmacol,2009,123(1):91-96
[28]林洪生,李树奇,裴迎霞,等.三参冲剂对肺癌转移中细胞内皮细胞及粘附因子的影响.1999,8(12)：574-576
[29]Cohen J. Apoptosis. Immunol Today,1993,14(3):126-130
[30]Son Y, Kim J, Chung Y, et al. Ripe fruits of Solanum nigrum L. inhibits cell growth and induces apoptosis in MCF-7 cells (J). Food chem toxicol,2003,41(10):1421-1428
[31]Friedman M, Fitch T, Yokoyama W. Lowering of plasma LDL cholesterol in hansters by the tomato glyco-alkaloid tomatine[J]. Food Chem Toxicol,2000,38(7):549-553
[32]季宇彬,万梅绪,高世勇,等.龙葵碱对荷瘤小鼠红细胞免疫功能的影响[J].中草药,2007,38(3)：412-414
[33]季宇彬,王胜惠,高世勇,等.龙葵碱对H22荷瘤小鼠肿瘤细胞膜唾液酸和封闭度的影响[J].中草药,2005,36(1)：79-81
[34]高世勇,王秋娟,季宇彬.龙葵碱对HepG2细胞内caspase-3及Bcl-2蛋白含量的影响[J].2006,4(3)：224-229
[35]高世勇,邹翔,王宏亮.龙葵碱诱导HepG2细胞凋亡的观察[J],2007,23(6)：644-650
[36]Lee K, Kozukue N, Han J, et al. Glycoalkaloid and metabolites inhibit the growth of human colon(HT29) and liver(HepG2) cancer cells[J]. Food Chem,2004,52 (10):2832-2839
[37]Heuvel M, Ingham P. Smoothened encodes a receptor-like serpentine protein required for hedgehog signalling[J]. Nature,1996,382(6591):547-551
[38]Taipale J, Chen JK, Cooper MK, et al. Effects of oncogenic mutations in Smoothened and Patched can be reversed by cyclopamine. Nature,2000,406(6799):1005-1009
[39]Markus D, Yasemin S, Antje H, et al. KAAD-cyclopamine augmentde TRAIL-mediated apoptosis in malignant glioma cells by modulating the intrinsic and extrinsic apoptotic pathway[J]. Neurobiol Disease,2009,34(2):259-266
[40]Hsiang Y, Hertzberg R, Hecht S, et al. Plant antitumor agents. Ⅰ. The isolation and structure of camptothecin, a novel alkaloidal leukemia and tumor inhibitor from Camptotheca acuminata[J]. J Am Chem Soc.1966,88(16):3888-3890
[41]Takimoto C, Wright J, Arbuck S, et al. Clinical applications of the camptothecins[J]. Biochim Biophys Acta.1998,1400(1-3):107-119
[42]姜琦,王海翔.喜树类药物的现代研究进展[J].药学进展,2007,31(9)：408-412
[43]Arakawa Y, Suzuki H, Saito S, et al. Novel missense mutation of the DNA topoisomerase Ⅰ gene in SN-38-resistant DLD-1 cells[J]. Mol Cancer Ther,2006,5(3):502-508
[44]Aisner J, Musanti R, Beers S, et al. Sequencing topotecan and etoposide plus cisplatin to overcome topoisomerase Ⅰ and Ⅱ resistance[J]. Clin Cancer Res,2003,9(7):2504-2509
[45]Miura H, Onishi H, Sasatsu M, et al. Antitumor characteristics of methoxypolyethylene glycol-poly(DL-lactic acid) nanoparticles containing camptothecin[J]. J Controlled Release,2004, 97(1):101-113
[46]Liu J, Li L, Cai Y, et al. Immobilization of camptothecin with surfactant into hydrogel for contrloode drug release[J]. Eur Polymer J,2006,42(8):1767-1774
[47]Le-Doan T, Perrouault L, Asseline U, et al. Recognition and photoinduced cleavage and cross-linking of nucleic acids by oligonucleotides covalently linked to ellipticine[J]. Antisense Res Dev,1991,1(1):43-45
[48]Jitka P, Tomas E, Jan H, et al. The mechanism of cytotoxicity and DNA adduct formation by the anticancer drug ellipticine in human neuroblastoma cells[J]. Biocheml Pharmacol,2009,77(9): 1466-1479
[49]Mizumoto K, SatoN, Kusumoto M, et al. Diverse effects of 9-hydroxyellipticine on the chemosensiticity of human pancreatic cancer cells harborin p53 mutations[J]. Cancer Lett,2000, 149(1-2):85-94
[50]Yung H, Woo Y, Su H, et al. Anti-invasive activity of sanguinarine through modulation of tight junctions and matrix metalloproteinase activities in MDA-MB-231 human breast carcinoma cells[J]. Chemico-Biological Interactions.2009,179(2-3):185-191
[51]Manu K, Girija K. Anti-metastatic potential of Punarnavine, an alkaloid from Boerhaavia diffusa Linn[J]. Immunobiol.2009,214(4):245-255
[52]顾瑞琦.几种十字花科植物幼芽提取液对元麦和小麦萌发种子的辐射保护作用[J].植物生理学报,1983,9(3)：307-316
[53]顾瑞琦.十字花科植物中的芥子碱对大麦和小麦萌发种子的辐射保护作用[J].中国科学(B辑),1986,2：146-152
[54]张明发,沈雅琴.芥子碱的抗炎和抗腹泻作用[J].中药药理与临床,1996(1)：29-31
[55]李群,顾瑞琦.十字花科植物中的芥子碱对果蝇的抗衰老作用[J].应用与环境生物学报,1999,5(1)：32-35
[56]李晶,石卓,赵丽娟,等.芥子碱硫酸氢盐对自发性高血压大鼠血压和心率的影响[J].吉林中医药,2002,22(6)：55-56
[57]吴国欣,林跃鑫,欧敏锐,等.芥子碱的抗雄激素作用[J].中国医药学报,2003,18(3)：142-144
[58]柯木根,吴国欣,陈密玉,等.芥子碱对鸡胚绒毛尿囊膜血管生成的影响[J].中华中医药杂志,2007,22(11)：36-39
[59]吴国欣,柯木根,詹烨.应用盘状血管生成模型评价芥子碱的抑制血管生成作用[J].海峡药学,2007,19(10)：22-24
[60]郭清龙.肿瘤药理学[M].化学工业出版社.北京,2007,12：51-52
[61]Kobayashi J, Suzuka M, et al. Konbuacidin, a new bromopoyrrole alkaloids with Cdk4 inhibitory activity from Hymeniacidon sponge. Tetrahedron,1997,53(46):15681
[62]Lock E, Reed C, Kinsey G, et al. Caspase-dependent and independent induction of phosphatidylserine externalization during apoptosis in human renal carcinoma Cak(1)-1 and A-498 cells. Toxicol,2007,229(1-2):79-90
[63]杨银峰,李家林.细胞凋亡相关基因与人类肿瘤.生命的化学,2004,24(3)：270-273
[64]刘瑛琪,冯媛.利用荧光探针JC-1检测心肌细胞线粒体膜电位的改变.解放军医学杂志,2002,27(8)：716-718
[65]Kim H, Kim E, Yang S, et al. Trichostatin A induces apoptosis in lung cancer cells via simultaneous activation of the death receptor-mediated and mitochondrial pathway? Exp Mol Med,2006,38(6):616-624
[66]宋泽庆,黎金庆.Fas/FasL介导免疫细胞凋亡与肿瘤免疫逃逸.医学综述,2000,6(7)：319-320
[67]Wang J. DNA topoisomerase[J]. Annu Rev Biochem,1996,65:635-692
[68]Liu L, Desai S, Li T, et al. Mechanism of action of camptothecin[J]. Ann N Y Acad Sci,2000, 922(1):1-10
[69]Ulukan H, Swaan PW. Camptothecins:A review of their chemotherapeutic potential[J]. Drugs, 2002,62(14):2039-2057
[70]Bogenrieder T, Herlyn M. Axis of evil:molecular mechanisms of cancer metastasis[J]. Oncogene, 2003,22(42):6524-6536
[71]Fidler I. The organ microenvionment and cancer metastasis [J]. Differentiation,2002,70(9-10): 498-505
[72]Fidler I, Tadinsky R. Genetic control of cancer metastasis[J]. J Natl Cancer Inst,1990,82(3): 166-168
[73]Hartsough M, Morrison D, Salerno M, et al. Nm23-H1 metastasis suppresor phosphorylation of kinase suppressor of Ras via a histidine protein kinase pathway[J]. J Biol Chem,2002,277(35): 32389-32399
[74]Ouatas T, Halcerson D, Steeg P. Dexamethasone and medroxyprogesterone acetate elevate Nm23-H1 metastasis suppressor gene expression in metastatic human breast carcinoma cells:new uses for old compounds[J], Clin Cancer Res,2003,9(10):3763-3772
[75]Wu Q, Ji Y, Zhang M, et al. Role of tumor metastasis suppressor gene KAI1 in digestive tract carcinomas and cancer cells [J]. Cell Tissue Res,2003,314(2):237-249
[76]Seraj M, Samant R, Goldberg S, et al. Functional evidence for human melanoma metastasis by the metastasis suppressor gene, BRMS1[J]. Exp Cell Res,2002:229-239
[77]Ozaki I, Mizuta T, Zhao G, et al. Involvement of the Ets-1 gene in overexpression of matrilysin in human hepatocellular carcinoma. Cancer Res,2000,60(22):6519-6525
[78]Kozlowska H, Naczk M, Shahidi F, et al. Canola and rapeseed:Production, chemistry, nutrition and processing technology [J]. USA:Van Nostrand Reinhold,1990:193-210
[79]Liu L, Wang Y, Li H, et al. Study of distribution of sinapine in commonly used crude drugs from cruciferous plants [J]. Chin J Chromatog.2006,24(1):49-51
[80]张新勇,徐中南,张勇耀,等.三子养亲汤提取工艺研究[J],中成药,2004,26(6)：500-501
[81]Clandinin D. Effecet of sinapine, the bitter substances in rapeseed oil meal, on the growth of chickens[J], Poultry Sci.,1961,40:484-487
[82]林燕妮,陈密玉,吴国欣,等.芥子碱提取工艺的研究[J].海峡药学,2006,18(6)：15-18
[83]王晓玲,许丽,胡利华,等.正交试验法优选白芥子延胡索甘遂合并提取的工艺[J].解放军药学学报,2006,22(3)：187-189
[84]江苏新医学学院.中药大辞典[M].上册.上海：上海人民出版社.1977：716
[85]张振凌,杨海玲,张本山,等.炮制对白芥子中芥子碱硫氰酸盐含量及煎出量的影响[J].中国中药杂志,2007,32(19)：2067-2069.
[86]梁燕,赖庆水,刘强.炒制对白芥子中芥子碱硫氰酸盐含量的影响[J].南方医科大学学报,2007,27(17)：1579-1580
[87]王维兰,朱进,莱萌,等.莱菔子浸膏片的试剂和质量控制[J].中成药研究,1987,5：3-4
[88]黄德娟,黄德超,梁桂燕.大孔树脂纯化花椰菜芥子碱的研究[J].食品研究与开发,2008,29(12)：15-17
[89]张力,金明宣,曹高娃.光谱实验室[J],1999,16(1)：51
[90]周同惠,徐礼.分析实验室[J],1990,9(4)：2
[91]Ferial I, Michael E. A new quantitative procedure for determination of sinapine[J], Agric Food chem,1979,27(4):917-918
[92]胡滨,陈一资,胡惠民.高效液相色谱法在食品快速检测中的应用[J].农产品加工,2007,2：33-35
[93]Liu L, Wang Y, Zhang X, et al. Determination of sinapine in semen raphani[J]. Chinese Trad Patent Med.2002,24(1):52-54
[94]杜松云,李其兰,余军.RP-HPLC法测定白芥子中芥子碱硫氰酸盐的含量[J].中国药房,2007,18(27)：2126-2128
[95]潘卫三,胡晋.薄层荧光扫描法及其在药物分析中的应用[J].现代应用药学,1990,7(4)：40-42
[96]王幕邹.药物分析杂志[J],1984,4(5)：314
[97]朱进,戴苏林.荧光扫描法测定莱菔子中芥子碱含量[J],中草药,1990,21(2)：13-15
[98]Tzagoloff A. Metabolism of sinapine in mustard paznts. I.Degradation of sinapine into sinapic acid and choline[J]. Plant Physiol,1963,38:202-206
[99]林燕妮,陈密玉,吴国欣,等.芥子碱含量测定方法探讨[J].药物分析杂志,2007,27(2)：260-263
[100]吴红星,杨剑婷.抗辐射的研究现状[J].放射免疫学杂志,2005,18(1)：58-60
[101]闫东,李莹.中医药抗衰老研究新进展[J].黑龙江中医药,2006,6：59-61
[102]郭房庆,李群,顾瑞琦.芥子碱对小鼠辐照后外周血象变化的影响[J].辐射研究与辐射工艺学报,1995,13(3)：177-180
[103]李卫业,李群, 郭房庆.辐射及活性氧对DNA的损伤以及芥子碱的保护作用[J].植物生理学报,1997,23(4)：319-323
[104]Rahman A, Choudhary M. Bioactive natural products as a potential source of new pharmacophores. a theory of memory[J]. Pure Appl Chem,2001,73:555
[105]何玲,李海涛,郭胜伟,等.芥子碱对大鼠脑匀浆和血清中乙酰胆碱酯酶的抑制作用[J].中国中药杂志,2008,33(7)：813-815
[106]张素玲.高血压防治的新观点、现状及对策[J].洛阳师专学报,2000,19(2)：142-144
[107]Folkman J. Tumor angiogenesis:therapeutic implications[J]. N Engl J Med,1971,285(21): 1182-1186
[108]Yang C, He L. Neuroprotective Effects of Sinapine on PC 12 Cells Apoptosis Induced by Sodium Dithionite[J]. Chin J Natural Med,2008,6:205-209
[109]田代华.实用中药辞典[M].北京：人民卫生出版社(上册):576
[110]刘明,张永萍,罗春丽.白芥子不同提取物对前列腺增生的作用[J].贵阳中医学院学报,2008,30(2)：15-17
[111]林燕妮,陈密玉,吴国欣等.芥子碱提取工艺的研究[J].海峡药学,2006,18(6)：15-18
[112]张振凌,杨海玲,张本山,等.炮制对白芥子中芥子碱硫氰酸盐含量及煎出量的影响.中国中药杂志,2007,32(19)：2067-2069
[113]Hui Z, Yinxi H, Tomonori H, et al. Electrochemistry of sinapine and its detection in medicinal plants[J]. Analtl Bioanalyt Chem,2005,382(4):1196-1201
[114]Cai R, Arntfield S. A rapid high-performance liquid chromatographic method for the determination of sinapine and sinapic acid in canola seed and meal[J]. J Am Oil Chemists' Society,2001,78:903-910
[115]Pawlikowski M, Melen M G. Perspectives of new potential therapeutic applications of somatostatin analogs [J]. Neuro Endocrinol Lett,2003,24(1-2):21-27
[116]Zhai Y, Lu Z J. Effect of thalidomide on tumor growth in mouse hepatoma H22 model[J]. Chinese journal of cancer,2003,22(12):1301-1306
[117]徐叔云,陈修.药理实验方法学[M].北京：人民卫生出版社(第三版).2005：1785-1788
[118]Hayon T, Dvilansky A, Shpilberg O, et al. Appraisal of the MTT-base assay as a useful tool for predicting drug chemosensitivity in leukemia[J]. Leukemia Lymphoma,2003,44(11):1957-1962
[119]Mosmann T. Rapid colorimetric assay for cellular growth and survival:application to proliferation and cytotoxicity assays[J]. Journal of Immunological Methods,1983,65(1-2):55-63
[120]Sargent J, Mitchell J, Degraff, et al Chemosensitivity testing of human lung cancer cell lines using the MTT assay[J]. Br J Cancer,1989,60(2):206-210
[121]刘建文.药理实验方法学——新技术与新方法[M].北京：化学工业出版社(第二版).2008,67-69
[122]姜勇男,李燕,陆虹,等.荷瘤小鼠免疫器官及免疫功能的研究.延边医学学院学报,1996,19(2)：70-72
[123]刘亚玲,刑欣,刑凌霄,等.杂色曲霉素对小鼠胸腺和脾脏转录因子Foxp3+调节性T淋巴细胞的影响.中国药理学与毒理学杂志,2009,6(23)：188-194
[124]高丽,张肇铭,杨官娥.光合细菌制剂对荷瘤小鼠免疫功能的影响及抑瘤作用.山西医科 大学学报,2004,35(1)：8-9
[125]郑国兴,范成成,冷波,等.文蛤多肽粉对人癌细胞SMMC-7721的抑制作用及免疫调节作用.厦门大学学报,2008,47(2)：135-137
[126]Wu X, Xi Z, Zhu M, et al. A Study of the Expressions of CD44V6、Bcl-2 and P53 for prediction of Pmgnisis in Adenomas and Carcinomas of Colorectum. Acta universitatis medicinalis secondae shanghai,2003,23(2):140-148
[127]O'Brien I, Reutelingsperger C, Holdaway K. The use of annexin-V and TUNEL to monitor the progression of apoptosis in plants. Cytomet,1997,29(1):28-35
[128]Harov V, Sabbah H, Shimoyama H, et al. Evidence of cardiocyte apoptosis in myo cardium of dogs with chronic heart failure [J]. Am J Pathol,1996,148(1):141-149.
[129]Sakakura C, Sweeney E A, Shirahama T, et al. Overexpression of bax sensitizes human breast cancer MCF-7 cells to radiation-induced apoptosis[J]. Int J Cancer,1996,67(1):101-105
[130]Tsujimoto Y, Shumizu S. Bcl-2 family:life-or-death switch. FEBS Letters,2000,466(1):6-10
[131]Griffiths G, Dudrez L, Morgan C, et al. Cell Damage-induced conformational changes of the pro-apoptotic protein Bak in vivo precede the onset of apoptosis. J Cell Biol,1999,144(5): 903-914
[132]Murphy K, Streips U, Lock R. Bcl-2 inhibits a Fas-induced confortnational change in the Bas N teminus and Bax mitochondrial translocation. J Biol Chem.2000,275(23):17225-17228
[133]Six S, MatsuyamaS, et al. The putative poreforming domain of Bax regulates mitochondrial localization and interaction with Bcl-xl. Mol Cell Biol.2000,20(5):1604-1615
[134]Gross A Jockel J, Wie M, et al. Enforced dimerization of BAX results in the translcation, mitochondnal dysfunction and apoptosis. EMBO J.1998,17:3878-3885
[2]Wani M, Taylor H, Wall M, et al. Plant antitumor agents. VI. The isolation and structure of taxol, a novel antileukemic and antitumor agent from Taxus brevifolia[J]. Am Chem Soc,1971,93(9): 2325-2327
[3]张骁,束梅英,张韬.紫杉醇的研究进展[J].中国医药技术与市场,2006,6(4)：40-46
[4]Stone R. Surprise! A fungus factory for taxol[J]? Science,1993,260(5150):154-155
[5]Markman M, Hall J, Spitz DW, et al. Phase II trial of weekly single-agent paclitaxel in platinum/paclitaxel-refractory ovarian cancer[J]. Clin Oncol,2002,20(9):2365-2369
[6]Alberto M, Frank F, Gabriel H, et al. Docetaxel for treatment of solid tumours:a systenmatic review of clinical data. Lancet Oncol,2005,6:229-239
[7]AL-Sarrf M. Treatment of locally advanced head and neck cancer[J]. Cancer Contr,2002,9(5): 387-399
[8]Schiff P, Fant J, Horwitz S. Promation of microtubule assembly in vitro by taxol. Nature 1979, 277(5698):665-667
[9]Scatena C, Stewart Z, Mays D. Mitotic phosphorylation of Bcl-2 during normal cell cycle progression and Paclitaxel-induced growth arest[J]. Biol Chem.1998,273(46):30777-30784
[10]Haldar S, Basu A, Croce C. Serine-70 is one of the critical sites for drug-induced Bcl-2 phosphorylation in cancer cells. Cancer Res.1998,58(8):1609-1615
[11]Panvichian R, Orth K, Day ML, et al. Paclitaxel-associated multiminricmcleation is permitted by the inhibition og caspas sctivation:a potential early step in druy resistance[J]. Cancer Res,1998, 58(20):4667
[12]孟刚,李庆林,陈志武,等.紫杉醇诱导MCF27凋亡与bcl-2及bax的关系[J].中国药理学通报,2001,17(3)：282-285
[13]Yoshino T, Shiina H, Urakami S, et al. Bcl-2 expression as a predictive marker of hormone-refractory prostate cancer treated with taxane-based chemotherapy. Clin Cancer Res, 2006,238(1):90-103
[14]Blagosklonny M, Schulte T, Nguyen P, et al. Taxol-induced apoptosis and phosphorylation of Bcl-2 protein involves c-Raf-1 and represents a novel c-Raf-1 signal transduction pathway[J]. Cancer Res,1996,56(8):1851-1854
[15]Uffelie OF. Alkaloids with tumor inhibiting action from Catharanthus roseus[J]. Pharm Weekbl, 1969,104(16):321-330
[16]Zhou X, Ranmani R. Preclinical pharmacology of vinca alkaloids[J]. Drugs,1992,44(4):1-16
[17]丁亚芳,包永明,安利佳.长春碱类抗肿瘤药物的研究进展[J].中国医药工业杂志,2005,36(7)：424-427
[18]Ngan VK, Bellman K, Panda D, et al. Novel actions of the antitumor drugs vinflunine and vinorelbine on microtubules[J], Cancer Res,2000,60:5045-5051
[19]Lobert S, Vulevic B, Correia J. Interaction of vinca alkaloids with tubulin:a comparison of vinblastine, vincristine, and vinorelbine[J]. Biochem,1996,35(21):6806-6814
[20]马彦.长春碱类抗肿瘤药物的研究进展.中山大学研究生学刊,2004,25(3)：30-39
[21]Pourroy B, Carre M, Honore S, et al. Low concentration of vinflunine induce apoptosis in human SK-N-SH neuroblastoma cells through a postmitotic G1 arrest and a mitochondrial pathway[J]. Mol Pharmacol,2004,66(3):580-591
[22]Azra R, Elham C, Zahra H. The effect of vinca alkaloid anticancer drug, vinorelbine, on chromatin and histone proteins in solution[J]. Eur. J. Pharmac,2009,613(1-3):34-38
[23]Yuan J, Zhang Z. Advances research and Bio-activity of matrine[J]. Pesticides,2003,42(7):1-4
[24]Wang T, Li s, Jian J, et al. Advances in the studies of anti-tumour activity of alkaloids in Radix Sophorae Flacescentis (Kushen)[J]. Chin J Exp Trad Med Form,2004,10(4):52-55
[25]Jin Y, Wu X, Lou J. Effects of matrine on cell proliferation, cell cycle and apoptosis of human hepatocarcinoma cells[J]. Chin. J. Clin Rehab.2006,10(3):107-109
[26]张鸣杰,黄健.苦参碱类抗肿瘤作用机制研究的新进展[J].中国中药杂志,2004,29(2)：115-118
[27]Dai Z, Gao J, Ji Z, et al. Matrine induces apoptosis in gastric carcinoma cells via alteration of Fas/FasL and activation of caspase-3[J]. J Ethnophrmacol,2009,123(1):91-96
[28]林洪生,李树奇,裴迎霞,等.三参冲剂对肺癌转移中细胞内皮细胞及粘附因子的影响.1999,8(12)：574-576
[29]Cohen J. Apoptosis. Immunol Today,1993,14(3):126-130
[30]Son Y, Kim J, Chung Y, et al. Ripe fruits of Solanum nigrum L. inhibits cell growth and induces apoptosis in MCF-7 cells (J). Food chem toxicol,2003,41(10):1421-1428
[31]Friedman M, Fitch T, Yokoyama W. Lowering of plasma LDL cholesterol in hansters by the tomato glyco-alkaloid tomatine[J]. Food Chem Toxicol,2000,38(7):549-553
[32]季宇彬,万梅绪,高世勇,等.龙葵碱对荷瘤小鼠红细胞免疫功能的影响[J].中草药,2007,38(3)：412-414
[33]季宇彬,王胜惠,高世勇,等.龙葵碱对H22荷瘤小鼠肿瘤细胞膜唾液酸和封闭度的影响[J].中草药,2005,36(1)：79-81
[34]高世勇,王秋娟,季宇彬.龙葵碱对HepG2细胞内caspase-3及Bcl-2蛋白含量的影响[J].2006,4(3)：224-229
[35]高世勇,邹翔,王宏亮.龙葵碱诱导HepG2细胞凋亡的观察[J],2007,23(6)：644-650
[36]Lee K, Kozukue N, Han J, et al. Glycoalkaloid and metabolites inhibit the growth of human colon(HT29) and liver(HepG2) cancer cells[J]. Food Chem,2004,52 (10):2832-2839
[37]Heuvel M, Ingham P. Smoothened encodes a receptor-like serpentine protein required for hedgehog signalling[J]. Nature,1996,382(6591):547-551
[38]Taipale J, Chen JK, Cooper MK, et al. Effects of oncogenic mutations in Smoothened and Patched can be reversed by cyclopamine. Nature,2000,406(6799):1005-1009
[39]Markus D, Yasemin S, Antje H, et al. KAAD-cyclopamine augmentde TRAIL-mediated apoptosis in malignant glioma cells by modulating the intrinsic and extrinsic apoptotic pathway[J]. Neurobiol Disease,2009,34(2):259-266
[40]Hsiang Y, Hertzberg R, Hecht S, et al. Plant antitumor agents. Ⅰ. The isolation and structure of camptothecin, a novel alkaloidal leukemia and tumor inhibitor from Camptotheca acuminata[J]. J Am Chem Soc.1966,88(16):3888-3890
[41]Takimoto C, Wright J, Arbuck S, et al. Clinical applications of the camptothecins[J]. Biochim Biophys Acta.1998,1400(1-3):107-119
[42]姜琦,王海翔.喜树类药物的现代研究进展[J].药学进展,2007,31(9)：408-412
[43]Arakawa Y, Suzuki H, Saito S, et al. Novel missense mutation of the DNA topoisomerase Ⅰ gene in SN-38-resistant DLD-1 cells[J]. Mol Cancer Ther,2006,5(3):502-508
[44]Aisner J, Musanti R, Beers S, et al. Sequencing topotecan and etoposide plus cisplatin to overcome topoisomerase Ⅰ and Ⅱ resistance[J]. Clin Cancer Res,2003,9(7):2504-2509
[45]Miura H, Onishi H, Sasatsu M, et al. Antitumor characteristics of methoxypolyethylene glycol-poly(DL-lactic acid) nanoparticles containing camptothecin[J]. J Controlled Release,2004, 97(1):101-113
[46]Liu J, Li L, Cai Y, et al. Immobilization of camptothecin with surfactant into hydrogel for contrloode drug release[J]. Eur Polymer J,2006,42(8):1767-1774
[47]Le-Doan T, Perrouault L, Asseline U, et al. Recognition and photoinduced cleavage and cross-linking of nucleic acids by oligonucleotides covalently linked to ellipticine[J]. Antisense Res Dev,1991,1(1):43-45
[48]Jitka P, Tomas E, Jan H, et al. The mechanism of cytotoxicity and DNA adduct formation by the anticancer drug ellipticine in human neuroblastoma cells[J]. Biocheml Pharmacol,2009,77(9): 1466-1479
[49]Mizumoto K, SatoN, Kusumoto M, et al. Diverse effects of 9-hydroxyellipticine on the chemosensiticity of human pancreatic cancer cells harborin p53 mutations[J]. Cancer Lett,2000, 149(1-2):85-94
[50]Yung H, Woo Y, Su H, et al. Anti-invasive activity of sanguinarine through modulation of tight junctions and matrix metalloproteinase activities in MDA-MB-231 human breast carcinoma cells[J]. Chemico-Biological Interactions.2009,179(2-3):185-191
[51]Manu K, Girija K. Anti-metastatic potential of Punarnavine, an alkaloid from Boerhaavia diffusa Linn[J]. Immunobiol.2009,214(4):245-255
[52]顾瑞琦.几种十字花科植物幼芽提取液对元麦和小麦萌发种子的辐射保护作用[J].植物生理学报,1983,9(3)：307-316
[53]顾瑞琦.十字花科植物中的芥子碱对大麦和小麦萌发种子的辐射保护作用[J].中国科学(B辑),1986,2：146-152
[54]张明发,沈雅琴.芥子碱的抗炎和抗腹泻作用[J].中药药理与临床,1996(1)：29-31
[55]李群,顾瑞琦.十字花科植物中的芥子碱对果蝇的抗衰老作用[J].应用与环境生物学报,1999,5(1)：32-35
[56]李晶,石卓,赵丽娟,等.芥子碱硫酸氢盐对自发性高血压大鼠血压和心率的影响[J].吉林中医药,2002,22(6)：55-56
[57]吴国欣,林跃鑫,欧敏锐,等.芥子碱的抗雄激素作用[J].中国医药学报,2003,18(3)：142-144
[58]柯木根,吴国欣,陈密玉,等.芥子碱对鸡胚绒毛尿囊膜血管生成的影响[J].中华中医药杂志,2007,22(11)：36-39
[59]吴国欣,柯木根,詹烨.应用盘状血管生成模型评价芥子碱的抑制血管生成作用[J].海峡药学,2007,19(10)：22-24
[60]郭清龙.肿瘤药理学[M].化学工业出版社.北京,2007,12：51-52
[61]Kobayashi J, Suzuka M, et al. Konbuacidin, a new bromopoyrrole alkaloids with Cdk4 inhibitory activity from Hymeniacidon sponge. Tetrahedron,1997,53(46):15681
[62]Lock E, Reed C, Kinsey G, et al. Caspase-dependent and independent induction of phosphatidylserine externalization during apoptosis in human renal carcinoma Cak(1)-1 and A-498 cells. Toxicol,2007,229(1-2):79-90
[63]杨银峰,李家林.细胞凋亡相关基因与人类肿瘤.生命的化学,2004,24(3)：270-273
[64]刘瑛琪,冯媛.利用荧光探针JC-1检测心肌细胞线粒体膜电位的改变.解放军医学杂志,2002,27(8)：716-718
[65]Kim H, Kim E, Yang S, et al. Trichostatin A induces apoptosis in lung cancer cells via simultaneous activation of the death receptor-mediated and mitochondrial pathway? Exp Mol Med,2006,38(6):616-624
[66]宋泽庆,黎金庆.Fas/FasL介导免疫细胞凋亡与肿瘤免疫逃逸.医学综述,2000,6(7)：319-320
[67]Wang J. DNA topoisomerase[J]. Annu Rev Biochem,1996,65:635-692
[68]Liu L, Desai S, Li T, et al. Mechanism of action of camptothecin[J]. Ann N Y Acad Sci,2000, 922(1):1-10
[69]Ulukan H, Swaan PW. Camptothecins:A review of their chemotherapeutic potential[J]. Drugs, 2002,62(14):2039-2057
[70]Bogenrieder T, Herlyn M. Axis of evil:molecular mechanisms of cancer metastasis[J]. Oncogene, 2003,22(42):6524-6536
[71]Fidler I. The organ microenvionment and cancer metastasis [J]. Differentiation,2002,70(9-10): 498-505
[72]Fidler I, Tadinsky R. Genetic control of cancer metastasis[J]. J Natl Cancer Inst,1990,82(3): 166-168
[73]Hartsough M, Morrison D, Salerno M, et al. Nm23-H1 metastasis suppresor phosphorylation of kinase suppressor of Ras via a histidine protein kinase pathway[J]. J Biol Chem,2002,277(35): 32389-32399
[74]Ouatas T, Halcerson D, Steeg P. Dexamethasone and medroxyprogesterone acetate elevate Nm23-H1 metastasis suppressor gene expression in metastatic human breast carcinoma cells:new uses for old compounds[J], Clin Cancer Res,2003,9(10):3763-3772
[75]Wu Q, Ji Y, Zhang M, et al. Role of tumor metastasis suppressor gene KAI1 in digestive tract carcinomas and cancer cells [J]. Cell Tissue Res,2003,314(2):237-249
[76]Seraj M, Samant R, Goldberg S, et al. Functional evidence for human melanoma metastasis by the metastasis suppressor gene, BRMS1[J]. Exp Cell Res,2002:229-239
[77]Ozaki I, Mizuta T, Zhao G, et al. Involvement of the Ets-1 gene in overexpression of matrilysin in human hepatocellular carcinoma. Cancer Res,2000,60(22):6519-6525
[78]Kozlowska H, Naczk M, Shahidi F, et al. Canola and rapeseed:Production, chemistry, nutrition and processing technology [J]. USA:Van Nostrand Reinhold,1990:193-210
[79]Liu L, Wang Y, Li H, et al. Study of distribution of sinapine in commonly used crude drugs from cruciferous plants [J]. Chin J Chromatog.2006,24(1):49-51
[80]张新勇,徐中南,张勇耀,等.三子养亲汤提取工艺研究[J],中成药,2004,26(6)：500-501
[81]Clandinin D. Effecet of sinapine, the bitter substances in rapeseed oil meal, on the growth of chickens[J], Poultry Sci.,1961,40:484-487
[82]林燕妮,陈密玉,吴国欣,等.芥子碱提取工艺的研究[J].海峡药学,2006,18(6)：15-18
[83]王晓玲,许丽,胡利华,等.正交试验法优选白芥子延胡索甘遂合并提取的工艺[J].解放军药学学报,2006,22(3)：187-189
[84]江苏新医学学院.中药大辞典[M].上册.上海：上海人民出版社.1977：716
[85]张振凌,杨海玲,张本山,等.炮制对白芥子中芥子碱硫氰酸盐含量及煎出量的影响[J].中国中药杂志,2007,32(19)：2067-2069.
[86]梁燕,赖庆水,刘强.炒制对白芥子中芥子碱硫氰酸盐含量的影响[J].南方医科大学学报,2007,27(17)：1579-1580
[87]王维兰,朱进,莱萌,等.莱菔子浸膏片的试剂和质量控制[J].中成药研究,1987,5：3-4
[88]黄德娟,黄德超,梁桂燕.大孔树脂纯化花椰菜芥子碱的研究[J].食品研究与开发,2008,29(12)：15-17
[89]张力,金明宣,曹高娃.光谱实验室[J],1999,16(1)：51
[90]周同惠,徐礼.分析实验室[J],1990,9(4)：2
[91]Ferial I, Michael E. A new quantitative procedure for determination of sinapine[J], Agric Food chem,1979,27(4):917-918
[92]胡滨,陈一资,胡惠民.高效液相色谱法在食品快速检测中的应用[J].农产品加工,2007,2：33-35
[93]Liu L, Wang Y, Zhang X, et al. Determination of sinapine in semen raphani[J]. Chinese Trad Patent Med.2002,24(1):52-54
[94]杜松云,李其兰,余军.RP-HPLC法测定白芥子中芥子碱硫氰酸盐的含量[J].中国药房,2007,18(27)：2126-2128
[95]潘卫三,胡晋.薄层荧光扫描法及其在药物分析中的应用[J].现代应用药学,1990,7(4)：40-42
[96]王幕邹.药物分析杂志[J],1984,4(5)：314
[97]朱进,戴苏林.荧光扫描法测定莱菔子中芥子碱含量[J],中草药,1990,21(2)：13-15
[98]Tzagoloff A. Metabolism of sinapine in mustard paznts. I.Degradation of sinapine into sinapic acid and choline[J]. Plant Physiol,1963,38:202-206
[99]林燕妮,陈密玉,吴国欣,等.芥子碱含量测定方法探讨[J].药物分析杂志,2007,27(2)：260-263
[100]吴红星,杨剑婷.抗辐射的研究现状[J].放射免疫学杂志,2005,18(1)：58-60
[101]闫东,李莹.中医药抗衰老研究新进展[J].黑龙江中医药,2006,6：59-61
[102]郭房庆,李群,顾瑞琦.芥子碱对小鼠辐照后外周血象变化的影响[J].辐射研究与辐射工艺学报,1995,13(3)：177-180
[103]李卫业,李群, 郭房庆.辐射及活性氧对DNA的损伤以及芥子碱的保护作用[J].植物生理学报,1997,23(4)：319-323
[104]Rahman A, Choudhary M. Bioactive natural products as a potential source of new pharmacophores. a theory of memory[J]. Pure Appl Chem,2001,73:555
[105]何玲,李海涛,郭胜伟,等.芥子碱对大鼠脑匀浆和血清中乙酰胆碱酯酶的抑制作用[J].中国中药杂志,2008,33(7)：813-815
[106]张素玲.高血压防治的新观点、现状及对策[J].洛阳师专学报,2000,19(2)：142-144
[107]Folkman J. Tumor angiogenesis:therapeutic implications[J]. N Engl J Med,1971,285(21): 1182-1186
[108]Yang C, He L. Neuroprotective Effects of Sinapine on PC 12 Cells Apoptosis Induced by Sodium Dithionite[J]. Chin J Natural Med,2008,6:205-209
[109]田代华.实用中药辞典[M].北京：人民卫生出版社(上册):576
[110]刘明,张永萍,罗春丽.白芥子不同提取物对前列腺增生的作用[J].贵阳中医学院学报,2008,30(2)：15-17
[111]林燕妮,陈密玉,吴国欣等.芥子碱提取工艺的研究[J].海峡药学,2006,18(6)：15-18
[112]张振凌,杨海玲,张本山,等.炮制对白芥子中芥子碱硫氰酸盐含量及煎出量的影响.中国中药杂志,2007,32(19)：2067-2069
[113]Hui Z, Yinxi H, Tomonori H, et al. Electrochemistry of sinapine and its detection in medicinal plants[J]. Analtl Bioanalyt Chem,2005,382(4):1196-1201
[114]Cai R, Arntfield S. A rapid high-performance liquid chromatographic method for the determination of sinapine and sinapic acid in canola seed and meal[J]. J Am Oil Chemists' Society,2001,78:903-910
[115]Pawlikowski M, Melen M G. Perspectives of new potential therapeutic applications of somatostatin analogs [J]. Neuro Endocrinol Lett,2003,24(1-2):21-27
[116]Zhai Y, Lu Z J. Effect of thalidomide on tumor growth in mouse hepatoma H22 model[J]. Chinese journal of cancer,2003,22(12):1301-1306
[117]徐叔云,陈修.药理实验方法学[M].北京：人民卫生出版社(第三版).2005：1785-1788
[118]Hayon T, Dvilansky A, Shpilberg O, et al. Appraisal of the MTT-base assay as a useful tool for predicting drug chemosensitivity in leukemia[J]. Leukemia Lymphoma,2003,44(11):1957-1962
[119]Mosmann T. Rapid colorimetric assay for cellular growth and survival:application to proliferation and cytotoxicity assays[J]. Journal of Immunological Methods,1983,65(1-2):55-63
[120]Sargent J, Mitchell J, Degraff, et al Chemosensitivity testing of human lung cancer cell lines using the MTT assay[J]. Br J Cancer,1989,60(2):206-210
[121]刘建文.药理实验方法学——新技术与新方法[M].北京：化学工业出版社(第二版).2008,67-69
[122]姜勇男,李燕,陆虹,等.荷瘤小鼠免疫器官及免疫功能的研究.延边医学学院学报,1996,19(2)：70-72
[123]刘亚玲,刑欣,刑凌霄,等.杂色曲霉素对小鼠胸腺和脾脏转录因子Foxp3+调节性T淋巴细胞的影响.中国药理学与毒理学杂志,2009,6(23)：188-194
[124]高丽,张肇铭,杨官娥.光合细菌制剂对荷瘤小鼠免疫功能的影响及抑瘤作用.山西医科 大学学报,2004,35(1)：8-9
[125]郑国兴,范成成,冷波,等.文蛤多肽粉对人癌细胞SMMC-7721的抑制作用及免疫调节作用.厦门大学学报,2008,47(2)：135-137
[126]Wu X, Xi Z, Zhu M, et al. A Study of the Expressions of CD44V6、Bcl-2 and P53 for prediction of Pmgnisis in Adenomas and Carcinomas of Colorectum. Acta universitatis medicinalis secondae shanghai,2003,23(2):140-148
[127]O'Brien I, Reutelingsperger C, Holdaway K. The use of annexin-V and TUNEL to monitor the progression of apoptosis in plants. Cytomet,1997,29(1):28-35
[128]Harov V, Sabbah H, Shimoyama H, et al. Evidence of cardiocyte apoptosis in myo cardium of dogs with chronic heart failure [J]. Am J Pathol,1996,148(1):141-149.
[129]Sakakura C, Sweeney E A, Shirahama T, et al. Overexpression of bax sensitizes human breast cancer MCF-7 cells to radiation-induced apoptosis[J]. Int J Cancer,1996,67(1):101-105
[130]Tsujimoto Y, Shumizu S. Bcl-2 family:life-or-death switch. FEBS Letters,2000,466(1):6-10
[131]Griffiths G, Dudrez L, Morgan C, et al. Cell Damage-induced conformational changes of the pro-apoptotic protein Bak in vivo precede the onset of apoptosis. J Cell Biol,1999,144(5): 903-914
[132]Murphy K, Streips U, Lock R. Bcl-2 inhibits a Fas-induced confortnational change in the Bas N teminus and Bax mitochondrial translocation. J Biol Chem.2000,275(23):17225-17228
[133]Six S, MatsuyamaS, et al. The putative poreforming domain of Bax regulates mitochondrial localization and interaction with Bcl-xl. Mol Cell Biol.2000,20(5):1604-1615
[134]Gross A Jockel J, Wie M, et al. Enforced dimerization of BAX results in the translcation, mitochondnal dysfunction and apoptosis. EMBO J.1998,17:3878-3885